transplantant rejection: experimental therapy with protein a
TRANSCRIPT
ACUTE AND CHRONIC
TRANSPLANT REJECTION
THERAPY WITH PROTEIN AELECTIVE IMMUNOSUPPRESSION
Dmitri Popov PhD Radiobiology MD (Russia)
Advanced Medical Technology and Systems Inc
Canada
Protein A
Acute and chronic transplant
rejection Therapy with protein A
Organ transplantation is the moving of an organ from one body to another or from a donor site to another location on the persons own body to replace the recipients damaged or absent organ The emerging field of regenerative medicine is allowing scientists and engineers to create organs to be re-grown from the persons own cells (stem cells or cells extracted from the failing organs)Organs andor tissues that are transplanted within the same persons body are called autografts Transplants that are recently performed between two subjects of the same species are called allografts Allografts can either be from a living or cadaveric source httpenwikipediaorgwikiOrgan_transplantation
Acute and chronic transplant
rejection Therapy with protein A
Organs that can be transplanted are the heart kidneys liver lungs pancreas intestine and thymus Tissues include bones tendons (both referred to as musculoskeletal grafts) cornea skin heart valves nerves and veins Worldwide the kidneys are the most commonly transplanted organs followed by the liver and then the heart Cornea and musculoskeletal grafts are the most commonly transplanted tissues these outnumber organ transplants by more than tenfold httpenwikipediaorgwikiOrgan_transplantation
Acute and chronic transplant
rejection Therapy with protein A
Transplant rejection is when transplanted tissue is rejected by the recipients immune system which destroys the transplanted tissue Transplant rejection can be lessened by determining the molecular similitude between donor and recipient and by use of immunosuppressant drugs after transplant httpenwikipediaorgwikiOrgan_transplantation
Acute and chronic transplant
rejection Therapy with protein A
Rejection is an adaptive immune response via cellular immunity (mediated by killer T cells inducing apoptosis of target cells) as well as humoral immunity (mediated by activated B cells secreting antibody molecules) though the action is joined by components of innate immune response (phagocytes and soluble immune proteins) Different types of transplanted tissues tend to favor different balances of rejection mechanisms httpenwikipediaorgwikiOrgan_transplantation
Acute and chronic transplant
rejection Therapy with protein A
Hyperacute rejection[edit]
Initiated by preexisting humoral
immunity hyperacute rejection manifests within
minutes after transplant and if tissue is left
implanted brings systemic inflammatory response
syndrome Of high risk in kidney transplants is
rapid clumping namely agglutination of red blood
cells (RBCs or erythrocytes) as an antibody
molecule binds multiple target cells at once
httpenwikipediaorgwikiOrgan_transplantation
Acute and chronic transplant
rejection Therapy with protein A
While kidneys can routinely be obtained from human donors most organs are in short supply leading to consideration of xenotransplants from other species Pigs are especially likely sources for xenotransplants chosen for the anatomical and physiological characteristics they share with humans However the sugar galactose-alpha-13-galactose(αGal) has been implicated as a major factor in hyperacute rejection in xenotransplantation Unlike virtually all other mammals humans and other primates do not make αGal and in fact recognize it as an antigen During transplantation xenoreactive natural antibodies recognize αGal on the graft endothelium as an antigen and the resulting complement-mediated immune response leads to a rejection of the transplant httpenwikipediaorgwikiOrgan_transplantation
Acute and chronic transplant
rejection Therapy with protein A
Acute rejection
Developing with formation of cellular immunity acute rejection occurs to some degree in all transplants except between identical twins unless immunosuppression is achieved (usually through drugs) Acute rejection begins as early as one week after transplant the risk being highest in the first three months though it can occur months to years later Highly vascular tissues such as kidney or liver often host the earliest signsmdashparticularly at endothelial cells lining blood vesselsmdashthough it eventually occurs in roughly 10 to 30 of liver transplants and 10 to 20 of kidney transplants httpenwikipediaorgwikiOrgan_transplantation
Acute and chronic transplant
rejection Therapy with protein A
A single episode of acute rejection can be recognized and promptly treated usually preventing organ failure but recurrent episodes lead to chronic rejection It is believed that the process of acute rejection is mediated by the cell mediated pathway specifically by mononuclear macrophages and T-lymphocytes httpenwikipediaorgwikiOrgan_transplantation
Acute and chronic transplant
rejection Therapy with protein A
Chronic Rejection
The term chronic rejection initially described long-term loss of function in transplanted organs via fibrosis of the transplanted tissues blood vessels This is now chronic allograft vasculopathy however leaving chronic rejection referring to rejection due to more patent aspects of immunity httpenwikipediaorgwikiOrgan_transplantation
Acute and chronic transplant
rejection Therapy with protein A
Chronic rejection explains long-term morbidity in most lung-transplant recipients the median survival roughly 47 years about half the span versus other major organ transplants[16] In histopathology the condition is bronchiolitisobliterans which clinically presents as progressive airflow obstruction often involving dyspnea and coughing and the patient eventually succumbs to pulmonary insufficiency or secondary acute infection
httpenwikipediaorgwikiOrgan_transplantation
Acute and chronic transplant
rejection Therapy with protein A
Airflow obstruction not ascribable to other cause is labeled bronchiolitis obliterans syndrome (BOS) confirmed by a persistent dropmdashthree or more weeksmdashin forced expiratory volume (FEV1) by at least 20[17] BOS is seen in over 50 of lung-transplant recipients by 5 years and in over 80 by ten years First noted is infiltration by lymphocytes followed by epithelial cell injury then inflammatory lesions and recruitment of fibroblasts and myofibroblasts which proliferate and secrete proteins forming scar tissue
Acute and chronic transplant
rejection Therapy with protein A
Generally thought unpredictable BOS
progression varies widely lung function may
suddenly fall but stabilize for years or rapidly
progress to death within a few months Risk
factors include prior acute rejection
episodes gastroesophageal reflux disease acute
infections particular age groups HLA mis-
matching lymphocytic bronchiolitis and graft
dysfunction (eg airway ischemia)
httpenwikipediaorgwikiOrgan_transplantation
Acute and chronic transplant
rejection Therapy with protein A
Immune Mechanisms of Transplant Rejections
Rejection is an adaptive immune response via cellular immunity (mediated by killer T cells inducing apoptosis of target cells) as well as humoral immunity (mediated by activated B cells secreting antibody molecules) though the action is joined by components of innate immune response (phagocytes and soluble immune proteins) Different types of transplanted tissues tend to favor different balances of rejection mechanisms
Acute and chronic transplant
rejection Therapy with protein A
Immunization
An animals exposure to the antigens of a different member of the same or similar species is allostimulation and the tissue is allogenic Transplanted organs are often acquired from a cadaver (usually a host who had succumbed to trauma) whose tissues had already sustained ischemia or inflammation
Dendritic cells (DCs) which are the primary antigen-presenting cells (APCs) of the donor tissue migrate to the recipients peripheral lymphoid tissue (lymphoid follicles and lymph nodes) and present the donors self peptides to the recipients lymphocytes (immune cells residing in lymphoid tissues) Lymphocytes include two classes that enact adaptive immunity also called specific immunity Lymphocytes of specific immunity T cellsmdashincluding the subclasses helper T cells and killer T cellsmdashand B cells
The recipients helper T cells coordinate specific immunity directed at the donors self peptides or at the donors Major histocompatibilitycomplex molecules or at both
Acute and chronic transplant
rejection Therapy with protein A
Immune memory
When memory helper T cells CD4 receptors bind to the MHC class II molecules which are expressed on the surfaces of the target cells of the graft tissue the memory helper T cells T cell receptors (TCRs) can recognize their target antigen that is presented by the MHC class II molecules The memory helper T cell subsequently produces clones that as effector cells secrete immune signalling molecules (cytokines) in approximately the cytokine balance that had prevailed at the memory helper T cells priming to memorize the antigen As the priming event in this instance occurred amid inflammation the immune memory is pro-inflammatory
Acute and chronic transplant
rejection Therapy with protein A
Cellular immunity
As a cell is indicated by the prefix cyto a cytotoxicinfluence destroys the cell Alloreactive killer T cells also called cytotoxic T lymphocytes (CTLs) have CD8 receptors that dock to the transplanted tissues MHC class I moleculeswhich display the donors self peptides (In the living donor such presentation of self antigens helped maintain self tolerance) Thereupon the T cell receptors (TCRs) of the killer T cells recognize their matching epitope and trigger the target cells programmed cell death by apoptosis
Acute and chronic transplant
rejection Therapy with protein A
Humoral immunity
Developed through an earlier primary exposure that primed specific immunity to the nonself antigen a transplant recipient can have specific antibody crossreacting with the donor tissue upon the transplant event a secondary exposure This is typical after earlier mismatching among ABO blood types during blood transfusion At this secondary exposure these crossreactive antibody molecules interact with aspects of innate immunitymdashsoluble immune proteins called complement and innate immune cells calledphagocytesmdashwhich inflames and destroys the transplanted tissue
Acute and chronic transplant
rejection Therapy with protein A
Antibody[edit]
Secreted by an activated B cell then
called plasma cell an antibody molecule is a
soluble immunoglobulin (Ig) whose basic unit is
shaped like the letter Y the two arms are theFab
regions while the single stalk is the Fc region
Each of the two tips of Fab region is the paratope
which binds a matching molecular sequence and
its 3D shape (conformation) altogether
called epitope within the target antigen
Acute and chronic transplant
rejection Therapy with protein A
Opsonization
The IgGs Fc region also enables opsonization by a phagocyte a process by which the Fc receptor on the phagocytemdashsuch as neutrophils in blood and macrophages in tissuesmdashbinds the antibody molecules FC stalk and the phagocyte exhibits enhanced uptake of the antigen attached to the antibody molecules Fab region
Acute and chronic transplant
rejection Therapy with protein A
Complement cascade
When the paratope of Ig class gamma (IgG) binds its matching epitope IgGs Fc region conformationally shifts and can host a complement protein initiating the complement cascade that terminates by punching a hole in a cell membrane With many holes so punched fluid rushes into the cell and ruptures it
Cell debris can be recognized as damage associated molecular patterns (DAMPs) by pattern recognition receptors (PRRs) such as Toll-like receptors (TLRs) on membranes ofphagocytes which thereupon secrete proinflammatory cytokines recruiting more phagocytes to traffic to the area by sensing the concentration gradient of the secreted cytokines (chemotaxis)
Acute and chronic transplant
rejection Therapy with protein A
Protein A is a
42 kDa surface protein originally found in the
cell wall of the bacterium Staphylococcus
aureus It is encoded by the spa gene and its
regulation is controlled by DNA topology
cellular osmolarity and a two-component
system called ArlS-ArlR It has found use in
biochemical research because of its ability to
bind immunoglobulins
Acute and chronic transplant
rejection Therapy with protein A
It is composed of five homologous Ig-binding domains that fold into a three-helix bundle Each domain is able to bind proteins from many mammalian species most notably IgGs It binds the heavy chain within the Fc region of most immunoglobulins and also within the Fab region in the case of the human VH3 family Through these interactions in serum where IgG molecules are bound in the wrong orientation (in relation to normal antibodyfunction) the bacteria disrupts opsonization and phagocytosis
Acute and chronic transplant
rejection Therapy with protein A
Purified (unconjugated) Thermo Scientific Pierce Recombinant Protein A is useful as the basis for preparing various kinds of probes or affinity media for detection or purification of rabbit and human antibodies especially IgGisotypes in immunoassays and antibody purification protocols
Features of Recombinant Protein A
bull Contains four Fc-binding domains per proteinbull Better than Protein G for polyclonal IgG from rabbit pig dog and cat serumbull Poorer than Protein G for mouse IgG1 human IgG3 and rat goat and cow antibodies
Acute and chronic transplant
rejection Therapy with protein A
Protein A is a cell wall component produced by several strains of Staphylococcus aureus that consists of a single polypeptide chain and contains little or no carbohydrate Recombinant Protein A is produced in E coli and functions essentially the same as native Protein A The Protein A molecule contains four high-affinity binding sites capable of interacting with the Fcregion from IgG of several species including human and rabbit Optimal binding occurs at pH 82 although binding is also effective at neutral or physiological conditions (pH 70 to 76)
Acute and chronic transplant
rejection Therapy with protein A
httpswwwlifetechnologiescomordercatalog
product21184
Acute and chronic transplant
rejection Therapy with protein A
httpstoolslifetechnologiescomcontentsfsm
anualsMAN0011438_Pierce_Ig_Bind_Protein
s_UGpdf
httpstoolslifetechnologiescomcontentsfsbr
ochuresTR0034-Ab-binding-proteinspdf
Acute and chronic transplant
rejection Therapy with protein A
Crystal structure of a Staphylococcus
aureus protein A domain complexed with
the Fab fragment of a human IgM antibody
Structural basis for recognition of B-cell
receptors and superantigen activity Marc Graille et al Proc Natl Acad Sci U S A
2000 May 9 97(10) 5399ndash5404 PMCID PMC25840
Immunology
Acute and chronic transplant
rejection Therapy with protein A
Staphylococcus aureus Protein A Promotes
Immune Suppression
Scott D Kobayashi
Frank R DeLeo
Acute and chronic transplant
rejection Therapy with protein A
SpA binds the Fc region of antibody and the Fabregions of the B-cell receptor processes that are known to block opsonophagocytosis and cause B-cell death in vitro In a recent study Falugi et al [F Falugi H K Kim D M Missiakas and O Schneewind mBio4(5)e00575-13 2013] showed that vaccination with spa mutant S aureus strains lacking antibody Fc-andor Fab-binding capacity protects against subsequent challenge with the USA300 epidemic strain The findings provide strong support for the idea that SpA promotes S aureus immune evasion in vivo and form the foundation for a new approach in our efforts to develop a vaccine that prevents severe S aureusinfections
Protein A
Acute and chronic transplant
rejection Therapy with protein A
Organ transplantation is the moving of an organ from one body to another or from a donor site to another location on the persons own body to replace the recipients damaged or absent organ The emerging field of regenerative medicine is allowing scientists and engineers to create organs to be re-grown from the persons own cells (stem cells or cells extracted from the failing organs)Organs andor tissues that are transplanted within the same persons body are called autografts Transplants that are recently performed between two subjects of the same species are called allografts Allografts can either be from a living or cadaveric source httpenwikipediaorgwikiOrgan_transplantation
Acute and chronic transplant
rejection Therapy with protein A
Organs that can be transplanted are the heart kidneys liver lungs pancreas intestine and thymus Tissues include bones tendons (both referred to as musculoskeletal grafts) cornea skin heart valves nerves and veins Worldwide the kidneys are the most commonly transplanted organs followed by the liver and then the heart Cornea and musculoskeletal grafts are the most commonly transplanted tissues these outnumber organ transplants by more than tenfold httpenwikipediaorgwikiOrgan_transplantation
Acute and chronic transplant
rejection Therapy with protein A
Transplant rejection is when transplanted tissue is rejected by the recipients immune system which destroys the transplanted tissue Transplant rejection can be lessened by determining the molecular similitude between donor and recipient and by use of immunosuppressant drugs after transplant httpenwikipediaorgwikiOrgan_transplantation
Acute and chronic transplant
rejection Therapy with protein A
Rejection is an adaptive immune response via cellular immunity (mediated by killer T cells inducing apoptosis of target cells) as well as humoral immunity (mediated by activated B cells secreting antibody molecules) though the action is joined by components of innate immune response (phagocytes and soluble immune proteins) Different types of transplanted tissues tend to favor different balances of rejection mechanisms httpenwikipediaorgwikiOrgan_transplantation
Acute and chronic transplant
rejection Therapy with protein A
Hyperacute rejection[edit]
Initiated by preexisting humoral
immunity hyperacute rejection manifests within
minutes after transplant and if tissue is left
implanted brings systemic inflammatory response
syndrome Of high risk in kidney transplants is
rapid clumping namely agglutination of red blood
cells (RBCs or erythrocytes) as an antibody
molecule binds multiple target cells at once
httpenwikipediaorgwikiOrgan_transplantation
Acute and chronic transplant
rejection Therapy with protein A
While kidneys can routinely be obtained from human donors most organs are in short supply leading to consideration of xenotransplants from other species Pigs are especially likely sources for xenotransplants chosen for the anatomical and physiological characteristics they share with humans However the sugar galactose-alpha-13-galactose(αGal) has been implicated as a major factor in hyperacute rejection in xenotransplantation Unlike virtually all other mammals humans and other primates do not make αGal and in fact recognize it as an antigen During transplantation xenoreactive natural antibodies recognize αGal on the graft endothelium as an antigen and the resulting complement-mediated immune response leads to a rejection of the transplant httpenwikipediaorgwikiOrgan_transplantation
Acute and chronic transplant
rejection Therapy with protein A
Acute rejection
Developing with formation of cellular immunity acute rejection occurs to some degree in all transplants except between identical twins unless immunosuppression is achieved (usually through drugs) Acute rejection begins as early as one week after transplant the risk being highest in the first three months though it can occur months to years later Highly vascular tissues such as kidney or liver often host the earliest signsmdashparticularly at endothelial cells lining blood vesselsmdashthough it eventually occurs in roughly 10 to 30 of liver transplants and 10 to 20 of kidney transplants httpenwikipediaorgwikiOrgan_transplantation
Acute and chronic transplant
rejection Therapy with protein A
A single episode of acute rejection can be recognized and promptly treated usually preventing organ failure but recurrent episodes lead to chronic rejection It is believed that the process of acute rejection is mediated by the cell mediated pathway specifically by mononuclear macrophages and T-lymphocytes httpenwikipediaorgwikiOrgan_transplantation
Acute and chronic transplant
rejection Therapy with protein A
Chronic Rejection
The term chronic rejection initially described long-term loss of function in transplanted organs via fibrosis of the transplanted tissues blood vessels This is now chronic allograft vasculopathy however leaving chronic rejection referring to rejection due to more patent aspects of immunity httpenwikipediaorgwikiOrgan_transplantation
Acute and chronic transplant
rejection Therapy with protein A
Chronic rejection explains long-term morbidity in most lung-transplant recipients the median survival roughly 47 years about half the span versus other major organ transplants[16] In histopathology the condition is bronchiolitisobliterans which clinically presents as progressive airflow obstruction often involving dyspnea and coughing and the patient eventually succumbs to pulmonary insufficiency or secondary acute infection
httpenwikipediaorgwikiOrgan_transplantation
Acute and chronic transplant
rejection Therapy with protein A
Airflow obstruction not ascribable to other cause is labeled bronchiolitis obliterans syndrome (BOS) confirmed by a persistent dropmdashthree or more weeksmdashin forced expiratory volume (FEV1) by at least 20[17] BOS is seen in over 50 of lung-transplant recipients by 5 years and in over 80 by ten years First noted is infiltration by lymphocytes followed by epithelial cell injury then inflammatory lesions and recruitment of fibroblasts and myofibroblasts which proliferate and secrete proteins forming scar tissue
Acute and chronic transplant
rejection Therapy with protein A
Generally thought unpredictable BOS
progression varies widely lung function may
suddenly fall but stabilize for years or rapidly
progress to death within a few months Risk
factors include prior acute rejection
episodes gastroesophageal reflux disease acute
infections particular age groups HLA mis-
matching lymphocytic bronchiolitis and graft
dysfunction (eg airway ischemia)
httpenwikipediaorgwikiOrgan_transplantation
Acute and chronic transplant
rejection Therapy with protein A
Immune Mechanisms of Transplant Rejections
Rejection is an adaptive immune response via cellular immunity (mediated by killer T cells inducing apoptosis of target cells) as well as humoral immunity (mediated by activated B cells secreting antibody molecules) though the action is joined by components of innate immune response (phagocytes and soluble immune proteins) Different types of transplanted tissues tend to favor different balances of rejection mechanisms
Acute and chronic transplant
rejection Therapy with protein A
Immunization
An animals exposure to the antigens of a different member of the same or similar species is allostimulation and the tissue is allogenic Transplanted organs are often acquired from a cadaver (usually a host who had succumbed to trauma) whose tissues had already sustained ischemia or inflammation
Dendritic cells (DCs) which are the primary antigen-presenting cells (APCs) of the donor tissue migrate to the recipients peripheral lymphoid tissue (lymphoid follicles and lymph nodes) and present the donors self peptides to the recipients lymphocytes (immune cells residing in lymphoid tissues) Lymphocytes include two classes that enact adaptive immunity also called specific immunity Lymphocytes of specific immunity T cellsmdashincluding the subclasses helper T cells and killer T cellsmdashand B cells
The recipients helper T cells coordinate specific immunity directed at the donors self peptides or at the donors Major histocompatibilitycomplex molecules or at both
Acute and chronic transplant
rejection Therapy with protein A
Immune memory
When memory helper T cells CD4 receptors bind to the MHC class II molecules which are expressed on the surfaces of the target cells of the graft tissue the memory helper T cells T cell receptors (TCRs) can recognize their target antigen that is presented by the MHC class II molecules The memory helper T cell subsequently produces clones that as effector cells secrete immune signalling molecules (cytokines) in approximately the cytokine balance that had prevailed at the memory helper T cells priming to memorize the antigen As the priming event in this instance occurred amid inflammation the immune memory is pro-inflammatory
Acute and chronic transplant
rejection Therapy with protein A
Cellular immunity
As a cell is indicated by the prefix cyto a cytotoxicinfluence destroys the cell Alloreactive killer T cells also called cytotoxic T lymphocytes (CTLs) have CD8 receptors that dock to the transplanted tissues MHC class I moleculeswhich display the donors self peptides (In the living donor such presentation of self antigens helped maintain self tolerance) Thereupon the T cell receptors (TCRs) of the killer T cells recognize their matching epitope and trigger the target cells programmed cell death by apoptosis
Acute and chronic transplant
rejection Therapy with protein A
Humoral immunity
Developed through an earlier primary exposure that primed specific immunity to the nonself antigen a transplant recipient can have specific antibody crossreacting with the donor tissue upon the transplant event a secondary exposure This is typical after earlier mismatching among ABO blood types during blood transfusion At this secondary exposure these crossreactive antibody molecules interact with aspects of innate immunitymdashsoluble immune proteins called complement and innate immune cells calledphagocytesmdashwhich inflames and destroys the transplanted tissue
Acute and chronic transplant
rejection Therapy with protein A
Antibody[edit]
Secreted by an activated B cell then
called plasma cell an antibody molecule is a
soluble immunoglobulin (Ig) whose basic unit is
shaped like the letter Y the two arms are theFab
regions while the single stalk is the Fc region
Each of the two tips of Fab region is the paratope
which binds a matching molecular sequence and
its 3D shape (conformation) altogether
called epitope within the target antigen
Acute and chronic transplant
rejection Therapy with protein A
Opsonization
The IgGs Fc region also enables opsonization by a phagocyte a process by which the Fc receptor on the phagocytemdashsuch as neutrophils in blood and macrophages in tissuesmdashbinds the antibody molecules FC stalk and the phagocyte exhibits enhanced uptake of the antigen attached to the antibody molecules Fab region
Acute and chronic transplant
rejection Therapy with protein A
Complement cascade
When the paratope of Ig class gamma (IgG) binds its matching epitope IgGs Fc region conformationally shifts and can host a complement protein initiating the complement cascade that terminates by punching a hole in a cell membrane With many holes so punched fluid rushes into the cell and ruptures it
Cell debris can be recognized as damage associated molecular patterns (DAMPs) by pattern recognition receptors (PRRs) such as Toll-like receptors (TLRs) on membranes ofphagocytes which thereupon secrete proinflammatory cytokines recruiting more phagocytes to traffic to the area by sensing the concentration gradient of the secreted cytokines (chemotaxis)
Acute and chronic transplant
rejection Therapy with protein A
Protein A is a
42 kDa surface protein originally found in the
cell wall of the bacterium Staphylococcus
aureus It is encoded by the spa gene and its
regulation is controlled by DNA topology
cellular osmolarity and a two-component
system called ArlS-ArlR It has found use in
biochemical research because of its ability to
bind immunoglobulins
Acute and chronic transplant
rejection Therapy with protein A
It is composed of five homologous Ig-binding domains that fold into a three-helix bundle Each domain is able to bind proteins from many mammalian species most notably IgGs It binds the heavy chain within the Fc region of most immunoglobulins and also within the Fab region in the case of the human VH3 family Through these interactions in serum where IgG molecules are bound in the wrong orientation (in relation to normal antibodyfunction) the bacteria disrupts opsonization and phagocytosis
Acute and chronic transplant
rejection Therapy with protein A
Purified (unconjugated) Thermo Scientific Pierce Recombinant Protein A is useful as the basis for preparing various kinds of probes or affinity media for detection or purification of rabbit and human antibodies especially IgGisotypes in immunoassays and antibody purification protocols
Features of Recombinant Protein A
bull Contains four Fc-binding domains per proteinbull Better than Protein G for polyclonal IgG from rabbit pig dog and cat serumbull Poorer than Protein G for mouse IgG1 human IgG3 and rat goat and cow antibodies
Acute and chronic transplant
rejection Therapy with protein A
Protein A is a cell wall component produced by several strains of Staphylococcus aureus that consists of a single polypeptide chain and contains little or no carbohydrate Recombinant Protein A is produced in E coli and functions essentially the same as native Protein A The Protein A molecule contains four high-affinity binding sites capable of interacting with the Fcregion from IgG of several species including human and rabbit Optimal binding occurs at pH 82 although binding is also effective at neutral or physiological conditions (pH 70 to 76)
Acute and chronic transplant
rejection Therapy with protein A
httpswwwlifetechnologiescomordercatalog
product21184
Acute and chronic transplant
rejection Therapy with protein A
httpstoolslifetechnologiescomcontentsfsm
anualsMAN0011438_Pierce_Ig_Bind_Protein
s_UGpdf
httpstoolslifetechnologiescomcontentsfsbr
ochuresTR0034-Ab-binding-proteinspdf
Acute and chronic transplant
rejection Therapy with protein A
Crystal structure of a Staphylococcus
aureus protein A domain complexed with
the Fab fragment of a human IgM antibody
Structural basis for recognition of B-cell
receptors and superantigen activity Marc Graille et al Proc Natl Acad Sci U S A
2000 May 9 97(10) 5399ndash5404 PMCID PMC25840
Immunology
Acute and chronic transplant
rejection Therapy with protein A
Staphylococcus aureus Protein A Promotes
Immune Suppression
Scott D Kobayashi
Frank R DeLeo
Acute and chronic transplant
rejection Therapy with protein A
SpA binds the Fc region of antibody and the Fabregions of the B-cell receptor processes that are known to block opsonophagocytosis and cause B-cell death in vitro In a recent study Falugi et al [F Falugi H K Kim D M Missiakas and O Schneewind mBio4(5)e00575-13 2013] showed that vaccination with spa mutant S aureus strains lacking antibody Fc-andor Fab-binding capacity protects against subsequent challenge with the USA300 epidemic strain The findings provide strong support for the idea that SpA promotes S aureus immune evasion in vivo and form the foundation for a new approach in our efforts to develop a vaccine that prevents severe S aureusinfections
Acute and chronic transplant
rejection Therapy with protein A
Organ transplantation is the moving of an organ from one body to another or from a donor site to another location on the persons own body to replace the recipients damaged or absent organ The emerging field of regenerative medicine is allowing scientists and engineers to create organs to be re-grown from the persons own cells (stem cells or cells extracted from the failing organs)Organs andor tissues that are transplanted within the same persons body are called autografts Transplants that are recently performed between two subjects of the same species are called allografts Allografts can either be from a living or cadaveric source httpenwikipediaorgwikiOrgan_transplantation
Acute and chronic transplant
rejection Therapy with protein A
Organs that can be transplanted are the heart kidneys liver lungs pancreas intestine and thymus Tissues include bones tendons (both referred to as musculoskeletal grafts) cornea skin heart valves nerves and veins Worldwide the kidneys are the most commonly transplanted organs followed by the liver and then the heart Cornea and musculoskeletal grafts are the most commonly transplanted tissues these outnumber organ transplants by more than tenfold httpenwikipediaorgwikiOrgan_transplantation
Acute and chronic transplant
rejection Therapy with protein A
Transplant rejection is when transplanted tissue is rejected by the recipients immune system which destroys the transplanted tissue Transplant rejection can be lessened by determining the molecular similitude between donor and recipient and by use of immunosuppressant drugs after transplant httpenwikipediaorgwikiOrgan_transplantation
Acute and chronic transplant
rejection Therapy with protein A
Rejection is an adaptive immune response via cellular immunity (mediated by killer T cells inducing apoptosis of target cells) as well as humoral immunity (mediated by activated B cells secreting antibody molecules) though the action is joined by components of innate immune response (phagocytes and soluble immune proteins) Different types of transplanted tissues tend to favor different balances of rejection mechanisms httpenwikipediaorgwikiOrgan_transplantation
Acute and chronic transplant
rejection Therapy with protein A
Hyperacute rejection[edit]
Initiated by preexisting humoral
immunity hyperacute rejection manifests within
minutes after transplant and if tissue is left
implanted brings systemic inflammatory response
syndrome Of high risk in kidney transplants is
rapid clumping namely agglutination of red blood
cells (RBCs or erythrocytes) as an antibody
molecule binds multiple target cells at once
httpenwikipediaorgwikiOrgan_transplantation
Acute and chronic transplant
rejection Therapy with protein A
While kidneys can routinely be obtained from human donors most organs are in short supply leading to consideration of xenotransplants from other species Pigs are especially likely sources for xenotransplants chosen for the anatomical and physiological characteristics they share with humans However the sugar galactose-alpha-13-galactose(αGal) has been implicated as a major factor in hyperacute rejection in xenotransplantation Unlike virtually all other mammals humans and other primates do not make αGal and in fact recognize it as an antigen During transplantation xenoreactive natural antibodies recognize αGal on the graft endothelium as an antigen and the resulting complement-mediated immune response leads to a rejection of the transplant httpenwikipediaorgwikiOrgan_transplantation
Acute and chronic transplant
rejection Therapy with protein A
Acute rejection
Developing with formation of cellular immunity acute rejection occurs to some degree in all transplants except between identical twins unless immunosuppression is achieved (usually through drugs) Acute rejection begins as early as one week after transplant the risk being highest in the first three months though it can occur months to years later Highly vascular tissues such as kidney or liver often host the earliest signsmdashparticularly at endothelial cells lining blood vesselsmdashthough it eventually occurs in roughly 10 to 30 of liver transplants and 10 to 20 of kidney transplants httpenwikipediaorgwikiOrgan_transplantation
Acute and chronic transplant
rejection Therapy with protein A
A single episode of acute rejection can be recognized and promptly treated usually preventing organ failure but recurrent episodes lead to chronic rejection It is believed that the process of acute rejection is mediated by the cell mediated pathway specifically by mononuclear macrophages and T-lymphocytes httpenwikipediaorgwikiOrgan_transplantation
Acute and chronic transplant
rejection Therapy with protein A
Chronic Rejection
The term chronic rejection initially described long-term loss of function in transplanted organs via fibrosis of the transplanted tissues blood vessels This is now chronic allograft vasculopathy however leaving chronic rejection referring to rejection due to more patent aspects of immunity httpenwikipediaorgwikiOrgan_transplantation
Acute and chronic transplant
rejection Therapy with protein A
Chronic rejection explains long-term morbidity in most lung-transplant recipients the median survival roughly 47 years about half the span versus other major organ transplants[16] In histopathology the condition is bronchiolitisobliterans which clinically presents as progressive airflow obstruction often involving dyspnea and coughing and the patient eventually succumbs to pulmonary insufficiency or secondary acute infection
httpenwikipediaorgwikiOrgan_transplantation
Acute and chronic transplant
rejection Therapy with protein A
Airflow obstruction not ascribable to other cause is labeled bronchiolitis obliterans syndrome (BOS) confirmed by a persistent dropmdashthree or more weeksmdashin forced expiratory volume (FEV1) by at least 20[17] BOS is seen in over 50 of lung-transplant recipients by 5 years and in over 80 by ten years First noted is infiltration by lymphocytes followed by epithelial cell injury then inflammatory lesions and recruitment of fibroblasts and myofibroblasts which proliferate and secrete proteins forming scar tissue
Acute and chronic transplant
rejection Therapy with protein A
Generally thought unpredictable BOS
progression varies widely lung function may
suddenly fall but stabilize for years or rapidly
progress to death within a few months Risk
factors include prior acute rejection
episodes gastroesophageal reflux disease acute
infections particular age groups HLA mis-
matching lymphocytic bronchiolitis and graft
dysfunction (eg airway ischemia)
httpenwikipediaorgwikiOrgan_transplantation
Acute and chronic transplant
rejection Therapy with protein A
Immune Mechanisms of Transplant Rejections
Rejection is an adaptive immune response via cellular immunity (mediated by killer T cells inducing apoptosis of target cells) as well as humoral immunity (mediated by activated B cells secreting antibody molecules) though the action is joined by components of innate immune response (phagocytes and soluble immune proteins) Different types of transplanted tissues tend to favor different balances of rejection mechanisms
Acute and chronic transplant
rejection Therapy with protein A
Immunization
An animals exposure to the antigens of a different member of the same or similar species is allostimulation and the tissue is allogenic Transplanted organs are often acquired from a cadaver (usually a host who had succumbed to trauma) whose tissues had already sustained ischemia or inflammation
Dendritic cells (DCs) which are the primary antigen-presenting cells (APCs) of the donor tissue migrate to the recipients peripheral lymphoid tissue (lymphoid follicles and lymph nodes) and present the donors self peptides to the recipients lymphocytes (immune cells residing in lymphoid tissues) Lymphocytes include two classes that enact adaptive immunity also called specific immunity Lymphocytes of specific immunity T cellsmdashincluding the subclasses helper T cells and killer T cellsmdashand B cells
The recipients helper T cells coordinate specific immunity directed at the donors self peptides or at the donors Major histocompatibilitycomplex molecules or at both
Acute and chronic transplant
rejection Therapy with protein A
Immune memory
When memory helper T cells CD4 receptors bind to the MHC class II molecules which are expressed on the surfaces of the target cells of the graft tissue the memory helper T cells T cell receptors (TCRs) can recognize their target antigen that is presented by the MHC class II molecules The memory helper T cell subsequently produces clones that as effector cells secrete immune signalling molecules (cytokines) in approximately the cytokine balance that had prevailed at the memory helper T cells priming to memorize the antigen As the priming event in this instance occurred amid inflammation the immune memory is pro-inflammatory
Acute and chronic transplant
rejection Therapy with protein A
Cellular immunity
As a cell is indicated by the prefix cyto a cytotoxicinfluence destroys the cell Alloreactive killer T cells also called cytotoxic T lymphocytes (CTLs) have CD8 receptors that dock to the transplanted tissues MHC class I moleculeswhich display the donors self peptides (In the living donor such presentation of self antigens helped maintain self tolerance) Thereupon the T cell receptors (TCRs) of the killer T cells recognize their matching epitope and trigger the target cells programmed cell death by apoptosis
Acute and chronic transplant
rejection Therapy with protein A
Humoral immunity
Developed through an earlier primary exposure that primed specific immunity to the nonself antigen a transplant recipient can have specific antibody crossreacting with the donor tissue upon the transplant event a secondary exposure This is typical after earlier mismatching among ABO blood types during blood transfusion At this secondary exposure these crossreactive antibody molecules interact with aspects of innate immunitymdashsoluble immune proteins called complement and innate immune cells calledphagocytesmdashwhich inflames and destroys the transplanted tissue
Acute and chronic transplant
rejection Therapy with protein A
Antibody[edit]
Secreted by an activated B cell then
called plasma cell an antibody molecule is a
soluble immunoglobulin (Ig) whose basic unit is
shaped like the letter Y the two arms are theFab
regions while the single stalk is the Fc region
Each of the two tips of Fab region is the paratope
which binds a matching molecular sequence and
its 3D shape (conformation) altogether
called epitope within the target antigen
Acute and chronic transplant
rejection Therapy with protein A
Opsonization
The IgGs Fc region also enables opsonization by a phagocyte a process by which the Fc receptor on the phagocytemdashsuch as neutrophils in blood and macrophages in tissuesmdashbinds the antibody molecules FC stalk and the phagocyte exhibits enhanced uptake of the antigen attached to the antibody molecules Fab region
Acute and chronic transplant
rejection Therapy with protein A
Complement cascade
When the paratope of Ig class gamma (IgG) binds its matching epitope IgGs Fc region conformationally shifts and can host a complement protein initiating the complement cascade that terminates by punching a hole in a cell membrane With many holes so punched fluid rushes into the cell and ruptures it
Cell debris can be recognized as damage associated molecular patterns (DAMPs) by pattern recognition receptors (PRRs) such as Toll-like receptors (TLRs) on membranes ofphagocytes which thereupon secrete proinflammatory cytokines recruiting more phagocytes to traffic to the area by sensing the concentration gradient of the secreted cytokines (chemotaxis)
Acute and chronic transplant
rejection Therapy with protein A
Protein A is a
42 kDa surface protein originally found in the
cell wall of the bacterium Staphylococcus
aureus It is encoded by the spa gene and its
regulation is controlled by DNA topology
cellular osmolarity and a two-component
system called ArlS-ArlR It has found use in
biochemical research because of its ability to
bind immunoglobulins
Acute and chronic transplant
rejection Therapy with protein A
It is composed of five homologous Ig-binding domains that fold into a three-helix bundle Each domain is able to bind proteins from many mammalian species most notably IgGs It binds the heavy chain within the Fc region of most immunoglobulins and also within the Fab region in the case of the human VH3 family Through these interactions in serum where IgG molecules are bound in the wrong orientation (in relation to normal antibodyfunction) the bacteria disrupts opsonization and phagocytosis
Acute and chronic transplant
rejection Therapy with protein A
Purified (unconjugated) Thermo Scientific Pierce Recombinant Protein A is useful as the basis for preparing various kinds of probes or affinity media for detection or purification of rabbit and human antibodies especially IgGisotypes in immunoassays and antibody purification protocols
Features of Recombinant Protein A
bull Contains four Fc-binding domains per proteinbull Better than Protein G for polyclonal IgG from rabbit pig dog and cat serumbull Poorer than Protein G for mouse IgG1 human IgG3 and rat goat and cow antibodies
Acute and chronic transplant
rejection Therapy with protein A
Protein A is a cell wall component produced by several strains of Staphylococcus aureus that consists of a single polypeptide chain and contains little or no carbohydrate Recombinant Protein A is produced in E coli and functions essentially the same as native Protein A The Protein A molecule contains four high-affinity binding sites capable of interacting with the Fcregion from IgG of several species including human and rabbit Optimal binding occurs at pH 82 although binding is also effective at neutral or physiological conditions (pH 70 to 76)
Acute and chronic transplant
rejection Therapy with protein A
httpswwwlifetechnologiescomordercatalog
product21184
Acute and chronic transplant
rejection Therapy with protein A
httpstoolslifetechnologiescomcontentsfsm
anualsMAN0011438_Pierce_Ig_Bind_Protein
s_UGpdf
httpstoolslifetechnologiescomcontentsfsbr
ochuresTR0034-Ab-binding-proteinspdf
Acute and chronic transplant
rejection Therapy with protein A
Crystal structure of a Staphylococcus
aureus protein A domain complexed with
the Fab fragment of a human IgM antibody
Structural basis for recognition of B-cell
receptors and superantigen activity Marc Graille et al Proc Natl Acad Sci U S A
2000 May 9 97(10) 5399ndash5404 PMCID PMC25840
Immunology
Acute and chronic transplant
rejection Therapy with protein A
Staphylococcus aureus Protein A Promotes
Immune Suppression
Scott D Kobayashi
Frank R DeLeo
Acute and chronic transplant
rejection Therapy with protein A
SpA binds the Fc region of antibody and the Fabregions of the B-cell receptor processes that are known to block opsonophagocytosis and cause B-cell death in vitro In a recent study Falugi et al [F Falugi H K Kim D M Missiakas and O Schneewind mBio4(5)e00575-13 2013] showed that vaccination with spa mutant S aureus strains lacking antibody Fc-andor Fab-binding capacity protects against subsequent challenge with the USA300 epidemic strain The findings provide strong support for the idea that SpA promotes S aureus immune evasion in vivo and form the foundation for a new approach in our efforts to develop a vaccine that prevents severe S aureusinfections
Acute and chronic transplant
rejection Therapy with protein A
Organs that can be transplanted are the heart kidneys liver lungs pancreas intestine and thymus Tissues include bones tendons (both referred to as musculoskeletal grafts) cornea skin heart valves nerves and veins Worldwide the kidneys are the most commonly transplanted organs followed by the liver and then the heart Cornea and musculoskeletal grafts are the most commonly transplanted tissues these outnumber organ transplants by more than tenfold httpenwikipediaorgwikiOrgan_transplantation
Acute and chronic transplant
rejection Therapy with protein A
Transplant rejection is when transplanted tissue is rejected by the recipients immune system which destroys the transplanted tissue Transplant rejection can be lessened by determining the molecular similitude between donor and recipient and by use of immunosuppressant drugs after transplant httpenwikipediaorgwikiOrgan_transplantation
Acute and chronic transplant
rejection Therapy with protein A
Rejection is an adaptive immune response via cellular immunity (mediated by killer T cells inducing apoptosis of target cells) as well as humoral immunity (mediated by activated B cells secreting antibody molecules) though the action is joined by components of innate immune response (phagocytes and soluble immune proteins) Different types of transplanted tissues tend to favor different balances of rejection mechanisms httpenwikipediaorgwikiOrgan_transplantation
Acute and chronic transplant
rejection Therapy with protein A
Hyperacute rejection[edit]
Initiated by preexisting humoral
immunity hyperacute rejection manifests within
minutes after transplant and if tissue is left
implanted brings systemic inflammatory response
syndrome Of high risk in kidney transplants is
rapid clumping namely agglutination of red blood
cells (RBCs or erythrocytes) as an antibody
molecule binds multiple target cells at once
httpenwikipediaorgwikiOrgan_transplantation
Acute and chronic transplant
rejection Therapy with protein A
While kidneys can routinely be obtained from human donors most organs are in short supply leading to consideration of xenotransplants from other species Pigs are especially likely sources for xenotransplants chosen for the anatomical and physiological characteristics they share with humans However the sugar galactose-alpha-13-galactose(αGal) has been implicated as a major factor in hyperacute rejection in xenotransplantation Unlike virtually all other mammals humans and other primates do not make αGal and in fact recognize it as an antigen During transplantation xenoreactive natural antibodies recognize αGal on the graft endothelium as an antigen and the resulting complement-mediated immune response leads to a rejection of the transplant httpenwikipediaorgwikiOrgan_transplantation
Acute and chronic transplant
rejection Therapy with protein A
Acute rejection
Developing with formation of cellular immunity acute rejection occurs to some degree in all transplants except between identical twins unless immunosuppression is achieved (usually through drugs) Acute rejection begins as early as one week after transplant the risk being highest in the first three months though it can occur months to years later Highly vascular tissues such as kidney or liver often host the earliest signsmdashparticularly at endothelial cells lining blood vesselsmdashthough it eventually occurs in roughly 10 to 30 of liver transplants and 10 to 20 of kidney transplants httpenwikipediaorgwikiOrgan_transplantation
Acute and chronic transplant
rejection Therapy with protein A
A single episode of acute rejection can be recognized and promptly treated usually preventing organ failure but recurrent episodes lead to chronic rejection It is believed that the process of acute rejection is mediated by the cell mediated pathway specifically by mononuclear macrophages and T-lymphocytes httpenwikipediaorgwikiOrgan_transplantation
Acute and chronic transplant
rejection Therapy with protein A
Chronic Rejection
The term chronic rejection initially described long-term loss of function in transplanted organs via fibrosis of the transplanted tissues blood vessels This is now chronic allograft vasculopathy however leaving chronic rejection referring to rejection due to more patent aspects of immunity httpenwikipediaorgwikiOrgan_transplantation
Acute and chronic transplant
rejection Therapy with protein A
Chronic rejection explains long-term morbidity in most lung-transplant recipients the median survival roughly 47 years about half the span versus other major organ transplants[16] In histopathology the condition is bronchiolitisobliterans which clinically presents as progressive airflow obstruction often involving dyspnea and coughing and the patient eventually succumbs to pulmonary insufficiency or secondary acute infection
httpenwikipediaorgwikiOrgan_transplantation
Acute and chronic transplant
rejection Therapy with protein A
Airflow obstruction not ascribable to other cause is labeled bronchiolitis obliterans syndrome (BOS) confirmed by a persistent dropmdashthree or more weeksmdashin forced expiratory volume (FEV1) by at least 20[17] BOS is seen in over 50 of lung-transplant recipients by 5 years and in over 80 by ten years First noted is infiltration by lymphocytes followed by epithelial cell injury then inflammatory lesions and recruitment of fibroblasts and myofibroblasts which proliferate and secrete proteins forming scar tissue
Acute and chronic transplant
rejection Therapy with protein A
Generally thought unpredictable BOS
progression varies widely lung function may
suddenly fall but stabilize for years or rapidly
progress to death within a few months Risk
factors include prior acute rejection
episodes gastroesophageal reflux disease acute
infections particular age groups HLA mis-
matching lymphocytic bronchiolitis and graft
dysfunction (eg airway ischemia)
httpenwikipediaorgwikiOrgan_transplantation
Acute and chronic transplant
rejection Therapy with protein A
Immune Mechanisms of Transplant Rejections
Rejection is an adaptive immune response via cellular immunity (mediated by killer T cells inducing apoptosis of target cells) as well as humoral immunity (mediated by activated B cells secreting antibody molecules) though the action is joined by components of innate immune response (phagocytes and soluble immune proteins) Different types of transplanted tissues tend to favor different balances of rejection mechanisms
Acute and chronic transplant
rejection Therapy with protein A
Immunization
An animals exposure to the antigens of a different member of the same or similar species is allostimulation and the tissue is allogenic Transplanted organs are often acquired from a cadaver (usually a host who had succumbed to trauma) whose tissues had already sustained ischemia or inflammation
Dendritic cells (DCs) which are the primary antigen-presenting cells (APCs) of the donor tissue migrate to the recipients peripheral lymphoid tissue (lymphoid follicles and lymph nodes) and present the donors self peptides to the recipients lymphocytes (immune cells residing in lymphoid tissues) Lymphocytes include two classes that enact adaptive immunity also called specific immunity Lymphocytes of specific immunity T cellsmdashincluding the subclasses helper T cells and killer T cellsmdashand B cells
The recipients helper T cells coordinate specific immunity directed at the donors self peptides or at the donors Major histocompatibilitycomplex molecules or at both
Acute and chronic transplant
rejection Therapy with protein A
Immune memory
When memory helper T cells CD4 receptors bind to the MHC class II molecules which are expressed on the surfaces of the target cells of the graft tissue the memory helper T cells T cell receptors (TCRs) can recognize their target antigen that is presented by the MHC class II molecules The memory helper T cell subsequently produces clones that as effector cells secrete immune signalling molecules (cytokines) in approximately the cytokine balance that had prevailed at the memory helper T cells priming to memorize the antigen As the priming event in this instance occurred amid inflammation the immune memory is pro-inflammatory
Acute and chronic transplant
rejection Therapy with protein A
Cellular immunity
As a cell is indicated by the prefix cyto a cytotoxicinfluence destroys the cell Alloreactive killer T cells also called cytotoxic T lymphocytes (CTLs) have CD8 receptors that dock to the transplanted tissues MHC class I moleculeswhich display the donors self peptides (In the living donor such presentation of self antigens helped maintain self tolerance) Thereupon the T cell receptors (TCRs) of the killer T cells recognize their matching epitope and trigger the target cells programmed cell death by apoptosis
Acute and chronic transplant
rejection Therapy with protein A
Humoral immunity
Developed through an earlier primary exposure that primed specific immunity to the nonself antigen a transplant recipient can have specific antibody crossreacting with the donor tissue upon the transplant event a secondary exposure This is typical after earlier mismatching among ABO blood types during blood transfusion At this secondary exposure these crossreactive antibody molecules interact with aspects of innate immunitymdashsoluble immune proteins called complement and innate immune cells calledphagocytesmdashwhich inflames and destroys the transplanted tissue
Acute and chronic transplant
rejection Therapy with protein A
Antibody[edit]
Secreted by an activated B cell then
called plasma cell an antibody molecule is a
soluble immunoglobulin (Ig) whose basic unit is
shaped like the letter Y the two arms are theFab
regions while the single stalk is the Fc region
Each of the two tips of Fab region is the paratope
which binds a matching molecular sequence and
its 3D shape (conformation) altogether
called epitope within the target antigen
Acute and chronic transplant
rejection Therapy with protein A
Opsonization
The IgGs Fc region also enables opsonization by a phagocyte a process by which the Fc receptor on the phagocytemdashsuch as neutrophils in blood and macrophages in tissuesmdashbinds the antibody molecules FC stalk and the phagocyte exhibits enhanced uptake of the antigen attached to the antibody molecules Fab region
Acute and chronic transplant
rejection Therapy with protein A
Complement cascade
When the paratope of Ig class gamma (IgG) binds its matching epitope IgGs Fc region conformationally shifts and can host a complement protein initiating the complement cascade that terminates by punching a hole in a cell membrane With many holes so punched fluid rushes into the cell and ruptures it
Cell debris can be recognized as damage associated molecular patterns (DAMPs) by pattern recognition receptors (PRRs) such as Toll-like receptors (TLRs) on membranes ofphagocytes which thereupon secrete proinflammatory cytokines recruiting more phagocytes to traffic to the area by sensing the concentration gradient of the secreted cytokines (chemotaxis)
Acute and chronic transplant
rejection Therapy with protein A
Protein A is a
42 kDa surface protein originally found in the
cell wall of the bacterium Staphylococcus
aureus It is encoded by the spa gene and its
regulation is controlled by DNA topology
cellular osmolarity and a two-component
system called ArlS-ArlR It has found use in
biochemical research because of its ability to
bind immunoglobulins
Acute and chronic transplant
rejection Therapy with protein A
It is composed of five homologous Ig-binding domains that fold into a three-helix bundle Each domain is able to bind proteins from many mammalian species most notably IgGs It binds the heavy chain within the Fc region of most immunoglobulins and also within the Fab region in the case of the human VH3 family Through these interactions in serum where IgG molecules are bound in the wrong orientation (in relation to normal antibodyfunction) the bacteria disrupts opsonization and phagocytosis
Acute and chronic transplant
rejection Therapy with protein A
Purified (unconjugated) Thermo Scientific Pierce Recombinant Protein A is useful as the basis for preparing various kinds of probes or affinity media for detection or purification of rabbit and human antibodies especially IgGisotypes in immunoassays and antibody purification protocols
Features of Recombinant Protein A
bull Contains four Fc-binding domains per proteinbull Better than Protein G for polyclonal IgG from rabbit pig dog and cat serumbull Poorer than Protein G for mouse IgG1 human IgG3 and rat goat and cow antibodies
Acute and chronic transplant
rejection Therapy with protein A
Protein A is a cell wall component produced by several strains of Staphylococcus aureus that consists of a single polypeptide chain and contains little or no carbohydrate Recombinant Protein A is produced in E coli and functions essentially the same as native Protein A The Protein A molecule contains four high-affinity binding sites capable of interacting with the Fcregion from IgG of several species including human and rabbit Optimal binding occurs at pH 82 although binding is also effective at neutral or physiological conditions (pH 70 to 76)
Acute and chronic transplant
rejection Therapy with protein A
httpswwwlifetechnologiescomordercatalog
product21184
Acute and chronic transplant
rejection Therapy with protein A
httpstoolslifetechnologiescomcontentsfsm
anualsMAN0011438_Pierce_Ig_Bind_Protein
s_UGpdf
httpstoolslifetechnologiescomcontentsfsbr
ochuresTR0034-Ab-binding-proteinspdf
Acute and chronic transplant
rejection Therapy with protein A
Crystal structure of a Staphylococcus
aureus protein A domain complexed with
the Fab fragment of a human IgM antibody
Structural basis for recognition of B-cell
receptors and superantigen activity Marc Graille et al Proc Natl Acad Sci U S A
2000 May 9 97(10) 5399ndash5404 PMCID PMC25840
Immunology
Acute and chronic transplant
rejection Therapy with protein A
Staphylococcus aureus Protein A Promotes
Immune Suppression
Scott D Kobayashi
Frank R DeLeo
Acute and chronic transplant
rejection Therapy with protein A
SpA binds the Fc region of antibody and the Fabregions of the B-cell receptor processes that are known to block opsonophagocytosis and cause B-cell death in vitro In a recent study Falugi et al [F Falugi H K Kim D M Missiakas and O Schneewind mBio4(5)e00575-13 2013] showed that vaccination with spa mutant S aureus strains lacking antibody Fc-andor Fab-binding capacity protects against subsequent challenge with the USA300 epidemic strain The findings provide strong support for the idea that SpA promotes S aureus immune evasion in vivo and form the foundation for a new approach in our efforts to develop a vaccine that prevents severe S aureusinfections
Acute and chronic transplant
rejection Therapy with protein A
Transplant rejection is when transplanted tissue is rejected by the recipients immune system which destroys the transplanted tissue Transplant rejection can be lessened by determining the molecular similitude between donor and recipient and by use of immunosuppressant drugs after transplant httpenwikipediaorgwikiOrgan_transplantation
Acute and chronic transplant
rejection Therapy with protein A
Rejection is an adaptive immune response via cellular immunity (mediated by killer T cells inducing apoptosis of target cells) as well as humoral immunity (mediated by activated B cells secreting antibody molecules) though the action is joined by components of innate immune response (phagocytes and soluble immune proteins) Different types of transplanted tissues tend to favor different balances of rejection mechanisms httpenwikipediaorgwikiOrgan_transplantation
Acute and chronic transplant
rejection Therapy with protein A
Hyperacute rejection[edit]
Initiated by preexisting humoral
immunity hyperacute rejection manifests within
minutes after transplant and if tissue is left
implanted brings systemic inflammatory response
syndrome Of high risk in kidney transplants is
rapid clumping namely agglutination of red blood
cells (RBCs or erythrocytes) as an antibody
molecule binds multiple target cells at once
httpenwikipediaorgwikiOrgan_transplantation
Acute and chronic transplant
rejection Therapy with protein A
While kidneys can routinely be obtained from human donors most organs are in short supply leading to consideration of xenotransplants from other species Pigs are especially likely sources for xenotransplants chosen for the anatomical and physiological characteristics they share with humans However the sugar galactose-alpha-13-galactose(αGal) has been implicated as a major factor in hyperacute rejection in xenotransplantation Unlike virtually all other mammals humans and other primates do not make αGal and in fact recognize it as an antigen During transplantation xenoreactive natural antibodies recognize αGal on the graft endothelium as an antigen and the resulting complement-mediated immune response leads to a rejection of the transplant httpenwikipediaorgwikiOrgan_transplantation
Acute and chronic transplant
rejection Therapy with protein A
Acute rejection
Developing with formation of cellular immunity acute rejection occurs to some degree in all transplants except between identical twins unless immunosuppression is achieved (usually through drugs) Acute rejection begins as early as one week after transplant the risk being highest in the first three months though it can occur months to years later Highly vascular tissues such as kidney or liver often host the earliest signsmdashparticularly at endothelial cells lining blood vesselsmdashthough it eventually occurs in roughly 10 to 30 of liver transplants and 10 to 20 of kidney transplants httpenwikipediaorgwikiOrgan_transplantation
Acute and chronic transplant
rejection Therapy with protein A
A single episode of acute rejection can be recognized and promptly treated usually preventing organ failure but recurrent episodes lead to chronic rejection It is believed that the process of acute rejection is mediated by the cell mediated pathway specifically by mononuclear macrophages and T-lymphocytes httpenwikipediaorgwikiOrgan_transplantation
Acute and chronic transplant
rejection Therapy with protein A
Chronic Rejection
The term chronic rejection initially described long-term loss of function in transplanted organs via fibrosis of the transplanted tissues blood vessels This is now chronic allograft vasculopathy however leaving chronic rejection referring to rejection due to more patent aspects of immunity httpenwikipediaorgwikiOrgan_transplantation
Acute and chronic transplant
rejection Therapy with protein A
Chronic rejection explains long-term morbidity in most lung-transplant recipients the median survival roughly 47 years about half the span versus other major organ transplants[16] In histopathology the condition is bronchiolitisobliterans which clinically presents as progressive airflow obstruction often involving dyspnea and coughing and the patient eventually succumbs to pulmonary insufficiency or secondary acute infection
httpenwikipediaorgwikiOrgan_transplantation
Acute and chronic transplant
rejection Therapy with protein A
Airflow obstruction not ascribable to other cause is labeled bronchiolitis obliterans syndrome (BOS) confirmed by a persistent dropmdashthree or more weeksmdashin forced expiratory volume (FEV1) by at least 20[17] BOS is seen in over 50 of lung-transplant recipients by 5 years and in over 80 by ten years First noted is infiltration by lymphocytes followed by epithelial cell injury then inflammatory lesions and recruitment of fibroblasts and myofibroblasts which proliferate and secrete proteins forming scar tissue
Acute and chronic transplant
rejection Therapy with protein A
Generally thought unpredictable BOS
progression varies widely lung function may
suddenly fall but stabilize for years or rapidly
progress to death within a few months Risk
factors include prior acute rejection
episodes gastroesophageal reflux disease acute
infections particular age groups HLA mis-
matching lymphocytic bronchiolitis and graft
dysfunction (eg airway ischemia)
httpenwikipediaorgwikiOrgan_transplantation
Acute and chronic transplant
rejection Therapy with protein A
Immune Mechanisms of Transplant Rejections
Rejection is an adaptive immune response via cellular immunity (mediated by killer T cells inducing apoptosis of target cells) as well as humoral immunity (mediated by activated B cells secreting antibody molecules) though the action is joined by components of innate immune response (phagocytes and soluble immune proteins) Different types of transplanted tissues tend to favor different balances of rejection mechanisms
Acute and chronic transplant
rejection Therapy with protein A
Immunization
An animals exposure to the antigens of a different member of the same or similar species is allostimulation and the tissue is allogenic Transplanted organs are often acquired from a cadaver (usually a host who had succumbed to trauma) whose tissues had already sustained ischemia or inflammation
Dendritic cells (DCs) which are the primary antigen-presenting cells (APCs) of the donor tissue migrate to the recipients peripheral lymphoid tissue (lymphoid follicles and lymph nodes) and present the donors self peptides to the recipients lymphocytes (immune cells residing in lymphoid tissues) Lymphocytes include two classes that enact adaptive immunity also called specific immunity Lymphocytes of specific immunity T cellsmdashincluding the subclasses helper T cells and killer T cellsmdashand B cells
The recipients helper T cells coordinate specific immunity directed at the donors self peptides or at the donors Major histocompatibilitycomplex molecules or at both
Acute and chronic transplant
rejection Therapy with protein A
Immune memory
When memory helper T cells CD4 receptors bind to the MHC class II molecules which are expressed on the surfaces of the target cells of the graft tissue the memory helper T cells T cell receptors (TCRs) can recognize their target antigen that is presented by the MHC class II molecules The memory helper T cell subsequently produces clones that as effector cells secrete immune signalling molecules (cytokines) in approximately the cytokine balance that had prevailed at the memory helper T cells priming to memorize the antigen As the priming event in this instance occurred amid inflammation the immune memory is pro-inflammatory
Acute and chronic transplant
rejection Therapy with protein A
Cellular immunity
As a cell is indicated by the prefix cyto a cytotoxicinfluence destroys the cell Alloreactive killer T cells also called cytotoxic T lymphocytes (CTLs) have CD8 receptors that dock to the transplanted tissues MHC class I moleculeswhich display the donors self peptides (In the living donor such presentation of self antigens helped maintain self tolerance) Thereupon the T cell receptors (TCRs) of the killer T cells recognize their matching epitope and trigger the target cells programmed cell death by apoptosis
Acute and chronic transplant
rejection Therapy with protein A
Humoral immunity
Developed through an earlier primary exposure that primed specific immunity to the nonself antigen a transplant recipient can have specific antibody crossreacting with the donor tissue upon the transplant event a secondary exposure This is typical after earlier mismatching among ABO blood types during blood transfusion At this secondary exposure these crossreactive antibody molecules interact with aspects of innate immunitymdashsoluble immune proteins called complement and innate immune cells calledphagocytesmdashwhich inflames and destroys the transplanted tissue
Acute and chronic transplant
rejection Therapy with protein A
Antibody[edit]
Secreted by an activated B cell then
called plasma cell an antibody molecule is a
soluble immunoglobulin (Ig) whose basic unit is
shaped like the letter Y the two arms are theFab
regions while the single stalk is the Fc region
Each of the two tips of Fab region is the paratope
which binds a matching molecular sequence and
its 3D shape (conformation) altogether
called epitope within the target antigen
Acute and chronic transplant
rejection Therapy with protein A
Opsonization
The IgGs Fc region also enables opsonization by a phagocyte a process by which the Fc receptor on the phagocytemdashsuch as neutrophils in blood and macrophages in tissuesmdashbinds the antibody molecules FC stalk and the phagocyte exhibits enhanced uptake of the antigen attached to the antibody molecules Fab region
Acute and chronic transplant
rejection Therapy with protein A
Complement cascade
When the paratope of Ig class gamma (IgG) binds its matching epitope IgGs Fc region conformationally shifts and can host a complement protein initiating the complement cascade that terminates by punching a hole in a cell membrane With many holes so punched fluid rushes into the cell and ruptures it
Cell debris can be recognized as damage associated molecular patterns (DAMPs) by pattern recognition receptors (PRRs) such as Toll-like receptors (TLRs) on membranes ofphagocytes which thereupon secrete proinflammatory cytokines recruiting more phagocytes to traffic to the area by sensing the concentration gradient of the secreted cytokines (chemotaxis)
Acute and chronic transplant
rejection Therapy with protein A
Protein A is a
42 kDa surface protein originally found in the
cell wall of the bacterium Staphylococcus
aureus It is encoded by the spa gene and its
regulation is controlled by DNA topology
cellular osmolarity and a two-component
system called ArlS-ArlR It has found use in
biochemical research because of its ability to
bind immunoglobulins
Acute and chronic transplant
rejection Therapy with protein A
It is composed of five homologous Ig-binding domains that fold into a three-helix bundle Each domain is able to bind proteins from many mammalian species most notably IgGs It binds the heavy chain within the Fc region of most immunoglobulins and also within the Fab region in the case of the human VH3 family Through these interactions in serum where IgG molecules are bound in the wrong orientation (in relation to normal antibodyfunction) the bacteria disrupts opsonization and phagocytosis
Acute and chronic transplant
rejection Therapy with protein A
Purified (unconjugated) Thermo Scientific Pierce Recombinant Protein A is useful as the basis for preparing various kinds of probes or affinity media for detection or purification of rabbit and human antibodies especially IgGisotypes in immunoassays and antibody purification protocols
Features of Recombinant Protein A
bull Contains four Fc-binding domains per proteinbull Better than Protein G for polyclonal IgG from rabbit pig dog and cat serumbull Poorer than Protein G for mouse IgG1 human IgG3 and rat goat and cow antibodies
Acute and chronic transplant
rejection Therapy with protein A
Protein A is a cell wall component produced by several strains of Staphylococcus aureus that consists of a single polypeptide chain and contains little or no carbohydrate Recombinant Protein A is produced in E coli and functions essentially the same as native Protein A The Protein A molecule contains four high-affinity binding sites capable of interacting with the Fcregion from IgG of several species including human and rabbit Optimal binding occurs at pH 82 although binding is also effective at neutral or physiological conditions (pH 70 to 76)
Acute and chronic transplant
rejection Therapy with protein A
httpswwwlifetechnologiescomordercatalog
product21184
Acute and chronic transplant
rejection Therapy with protein A
httpstoolslifetechnologiescomcontentsfsm
anualsMAN0011438_Pierce_Ig_Bind_Protein
s_UGpdf
httpstoolslifetechnologiescomcontentsfsbr
ochuresTR0034-Ab-binding-proteinspdf
Acute and chronic transplant
rejection Therapy with protein A
Crystal structure of a Staphylococcus
aureus protein A domain complexed with
the Fab fragment of a human IgM antibody
Structural basis for recognition of B-cell
receptors and superantigen activity Marc Graille et al Proc Natl Acad Sci U S A
2000 May 9 97(10) 5399ndash5404 PMCID PMC25840
Immunology
Acute and chronic transplant
rejection Therapy with protein A
Staphylococcus aureus Protein A Promotes
Immune Suppression
Scott D Kobayashi
Frank R DeLeo
Acute and chronic transplant
rejection Therapy with protein A
SpA binds the Fc region of antibody and the Fabregions of the B-cell receptor processes that are known to block opsonophagocytosis and cause B-cell death in vitro In a recent study Falugi et al [F Falugi H K Kim D M Missiakas and O Schneewind mBio4(5)e00575-13 2013] showed that vaccination with spa mutant S aureus strains lacking antibody Fc-andor Fab-binding capacity protects against subsequent challenge with the USA300 epidemic strain The findings provide strong support for the idea that SpA promotes S aureus immune evasion in vivo and form the foundation for a new approach in our efforts to develop a vaccine that prevents severe S aureusinfections
Acute and chronic transplant
rejection Therapy with protein A
Rejection is an adaptive immune response via cellular immunity (mediated by killer T cells inducing apoptosis of target cells) as well as humoral immunity (mediated by activated B cells secreting antibody molecules) though the action is joined by components of innate immune response (phagocytes and soluble immune proteins) Different types of transplanted tissues tend to favor different balances of rejection mechanisms httpenwikipediaorgwikiOrgan_transplantation
Acute and chronic transplant
rejection Therapy with protein A
Hyperacute rejection[edit]
Initiated by preexisting humoral
immunity hyperacute rejection manifests within
minutes after transplant and if tissue is left
implanted brings systemic inflammatory response
syndrome Of high risk in kidney transplants is
rapid clumping namely agglutination of red blood
cells (RBCs or erythrocytes) as an antibody
molecule binds multiple target cells at once
httpenwikipediaorgwikiOrgan_transplantation
Acute and chronic transplant
rejection Therapy with protein A
While kidneys can routinely be obtained from human donors most organs are in short supply leading to consideration of xenotransplants from other species Pigs are especially likely sources for xenotransplants chosen for the anatomical and physiological characteristics they share with humans However the sugar galactose-alpha-13-galactose(αGal) has been implicated as a major factor in hyperacute rejection in xenotransplantation Unlike virtually all other mammals humans and other primates do not make αGal and in fact recognize it as an antigen During transplantation xenoreactive natural antibodies recognize αGal on the graft endothelium as an antigen and the resulting complement-mediated immune response leads to a rejection of the transplant httpenwikipediaorgwikiOrgan_transplantation
Acute and chronic transplant
rejection Therapy with protein A
Acute rejection
Developing with formation of cellular immunity acute rejection occurs to some degree in all transplants except between identical twins unless immunosuppression is achieved (usually through drugs) Acute rejection begins as early as one week after transplant the risk being highest in the first three months though it can occur months to years later Highly vascular tissues such as kidney or liver often host the earliest signsmdashparticularly at endothelial cells lining blood vesselsmdashthough it eventually occurs in roughly 10 to 30 of liver transplants and 10 to 20 of kidney transplants httpenwikipediaorgwikiOrgan_transplantation
Acute and chronic transplant
rejection Therapy with protein A
A single episode of acute rejection can be recognized and promptly treated usually preventing organ failure but recurrent episodes lead to chronic rejection It is believed that the process of acute rejection is mediated by the cell mediated pathway specifically by mononuclear macrophages and T-lymphocytes httpenwikipediaorgwikiOrgan_transplantation
Acute and chronic transplant
rejection Therapy with protein A
Chronic Rejection
The term chronic rejection initially described long-term loss of function in transplanted organs via fibrosis of the transplanted tissues blood vessels This is now chronic allograft vasculopathy however leaving chronic rejection referring to rejection due to more patent aspects of immunity httpenwikipediaorgwikiOrgan_transplantation
Acute and chronic transplant
rejection Therapy with protein A
Chronic rejection explains long-term morbidity in most lung-transplant recipients the median survival roughly 47 years about half the span versus other major organ transplants[16] In histopathology the condition is bronchiolitisobliterans which clinically presents as progressive airflow obstruction often involving dyspnea and coughing and the patient eventually succumbs to pulmonary insufficiency or secondary acute infection
httpenwikipediaorgwikiOrgan_transplantation
Acute and chronic transplant
rejection Therapy with protein A
Airflow obstruction not ascribable to other cause is labeled bronchiolitis obliterans syndrome (BOS) confirmed by a persistent dropmdashthree or more weeksmdashin forced expiratory volume (FEV1) by at least 20[17] BOS is seen in over 50 of lung-transplant recipients by 5 years and in over 80 by ten years First noted is infiltration by lymphocytes followed by epithelial cell injury then inflammatory lesions and recruitment of fibroblasts and myofibroblasts which proliferate and secrete proteins forming scar tissue
Acute and chronic transplant
rejection Therapy with protein A
Generally thought unpredictable BOS
progression varies widely lung function may
suddenly fall but stabilize for years or rapidly
progress to death within a few months Risk
factors include prior acute rejection
episodes gastroesophageal reflux disease acute
infections particular age groups HLA mis-
matching lymphocytic bronchiolitis and graft
dysfunction (eg airway ischemia)
httpenwikipediaorgwikiOrgan_transplantation
Acute and chronic transplant
rejection Therapy with protein A
Immune Mechanisms of Transplant Rejections
Rejection is an adaptive immune response via cellular immunity (mediated by killer T cells inducing apoptosis of target cells) as well as humoral immunity (mediated by activated B cells secreting antibody molecules) though the action is joined by components of innate immune response (phagocytes and soluble immune proteins) Different types of transplanted tissues tend to favor different balances of rejection mechanisms
Acute and chronic transplant
rejection Therapy with protein A
Immunization
An animals exposure to the antigens of a different member of the same or similar species is allostimulation and the tissue is allogenic Transplanted organs are often acquired from a cadaver (usually a host who had succumbed to trauma) whose tissues had already sustained ischemia or inflammation
Dendritic cells (DCs) which are the primary antigen-presenting cells (APCs) of the donor tissue migrate to the recipients peripheral lymphoid tissue (lymphoid follicles and lymph nodes) and present the donors self peptides to the recipients lymphocytes (immune cells residing in lymphoid tissues) Lymphocytes include two classes that enact adaptive immunity also called specific immunity Lymphocytes of specific immunity T cellsmdashincluding the subclasses helper T cells and killer T cellsmdashand B cells
The recipients helper T cells coordinate specific immunity directed at the donors self peptides or at the donors Major histocompatibilitycomplex molecules or at both
Acute and chronic transplant
rejection Therapy with protein A
Immune memory
When memory helper T cells CD4 receptors bind to the MHC class II molecules which are expressed on the surfaces of the target cells of the graft tissue the memory helper T cells T cell receptors (TCRs) can recognize their target antigen that is presented by the MHC class II molecules The memory helper T cell subsequently produces clones that as effector cells secrete immune signalling molecules (cytokines) in approximately the cytokine balance that had prevailed at the memory helper T cells priming to memorize the antigen As the priming event in this instance occurred amid inflammation the immune memory is pro-inflammatory
Acute and chronic transplant
rejection Therapy with protein A
Cellular immunity
As a cell is indicated by the prefix cyto a cytotoxicinfluence destroys the cell Alloreactive killer T cells also called cytotoxic T lymphocytes (CTLs) have CD8 receptors that dock to the transplanted tissues MHC class I moleculeswhich display the donors self peptides (In the living donor such presentation of self antigens helped maintain self tolerance) Thereupon the T cell receptors (TCRs) of the killer T cells recognize their matching epitope and trigger the target cells programmed cell death by apoptosis
Acute and chronic transplant
rejection Therapy with protein A
Humoral immunity
Developed through an earlier primary exposure that primed specific immunity to the nonself antigen a transplant recipient can have specific antibody crossreacting with the donor tissue upon the transplant event a secondary exposure This is typical after earlier mismatching among ABO blood types during blood transfusion At this secondary exposure these crossreactive antibody molecules interact with aspects of innate immunitymdashsoluble immune proteins called complement and innate immune cells calledphagocytesmdashwhich inflames and destroys the transplanted tissue
Acute and chronic transplant
rejection Therapy with protein A
Antibody[edit]
Secreted by an activated B cell then
called plasma cell an antibody molecule is a
soluble immunoglobulin (Ig) whose basic unit is
shaped like the letter Y the two arms are theFab
regions while the single stalk is the Fc region
Each of the two tips of Fab region is the paratope
which binds a matching molecular sequence and
its 3D shape (conformation) altogether
called epitope within the target antigen
Acute and chronic transplant
rejection Therapy with protein A
Opsonization
The IgGs Fc region also enables opsonization by a phagocyte a process by which the Fc receptor on the phagocytemdashsuch as neutrophils in blood and macrophages in tissuesmdashbinds the antibody molecules FC stalk and the phagocyte exhibits enhanced uptake of the antigen attached to the antibody molecules Fab region
Acute and chronic transplant
rejection Therapy with protein A
Complement cascade
When the paratope of Ig class gamma (IgG) binds its matching epitope IgGs Fc region conformationally shifts and can host a complement protein initiating the complement cascade that terminates by punching a hole in a cell membrane With many holes so punched fluid rushes into the cell and ruptures it
Cell debris can be recognized as damage associated molecular patterns (DAMPs) by pattern recognition receptors (PRRs) such as Toll-like receptors (TLRs) on membranes ofphagocytes which thereupon secrete proinflammatory cytokines recruiting more phagocytes to traffic to the area by sensing the concentration gradient of the secreted cytokines (chemotaxis)
Acute and chronic transplant
rejection Therapy with protein A
Protein A is a
42 kDa surface protein originally found in the
cell wall of the bacterium Staphylococcus
aureus It is encoded by the spa gene and its
regulation is controlled by DNA topology
cellular osmolarity and a two-component
system called ArlS-ArlR It has found use in
biochemical research because of its ability to
bind immunoglobulins
Acute and chronic transplant
rejection Therapy with protein A
It is composed of five homologous Ig-binding domains that fold into a three-helix bundle Each domain is able to bind proteins from many mammalian species most notably IgGs It binds the heavy chain within the Fc region of most immunoglobulins and also within the Fab region in the case of the human VH3 family Through these interactions in serum where IgG molecules are bound in the wrong orientation (in relation to normal antibodyfunction) the bacteria disrupts opsonization and phagocytosis
Acute and chronic transplant
rejection Therapy with protein A
Purified (unconjugated) Thermo Scientific Pierce Recombinant Protein A is useful as the basis for preparing various kinds of probes or affinity media for detection or purification of rabbit and human antibodies especially IgGisotypes in immunoassays and antibody purification protocols
Features of Recombinant Protein A
bull Contains four Fc-binding domains per proteinbull Better than Protein G for polyclonal IgG from rabbit pig dog and cat serumbull Poorer than Protein G for mouse IgG1 human IgG3 and rat goat and cow antibodies
Acute and chronic transplant
rejection Therapy with protein A
Protein A is a cell wall component produced by several strains of Staphylococcus aureus that consists of a single polypeptide chain and contains little or no carbohydrate Recombinant Protein A is produced in E coli and functions essentially the same as native Protein A The Protein A molecule contains four high-affinity binding sites capable of interacting with the Fcregion from IgG of several species including human and rabbit Optimal binding occurs at pH 82 although binding is also effective at neutral or physiological conditions (pH 70 to 76)
Acute and chronic transplant
rejection Therapy with protein A
httpswwwlifetechnologiescomordercatalog
product21184
Acute and chronic transplant
rejection Therapy with protein A
httpstoolslifetechnologiescomcontentsfsm
anualsMAN0011438_Pierce_Ig_Bind_Protein
s_UGpdf
httpstoolslifetechnologiescomcontentsfsbr
ochuresTR0034-Ab-binding-proteinspdf
Acute and chronic transplant
rejection Therapy with protein A
Crystal structure of a Staphylococcus
aureus protein A domain complexed with
the Fab fragment of a human IgM antibody
Structural basis for recognition of B-cell
receptors and superantigen activity Marc Graille et al Proc Natl Acad Sci U S A
2000 May 9 97(10) 5399ndash5404 PMCID PMC25840
Immunology
Acute and chronic transplant
rejection Therapy with protein A
Staphylococcus aureus Protein A Promotes
Immune Suppression
Scott D Kobayashi
Frank R DeLeo
Acute and chronic transplant
rejection Therapy with protein A
SpA binds the Fc region of antibody and the Fabregions of the B-cell receptor processes that are known to block opsonophagocytosis and cause B-cell death in vitro In a recent study Falugi et al [F Falugi H K Kim D M Missiakas and O Schneewind mBio4(5)e00575-13 2013] showed that vaccination with spa mutant S aureus strains lacking antibody Fc-andor Fab-binding capacity protects against subsequent challenge with the USA300 epidemic strain The findings provide strong support for the idea that SpA promotes S aureus immune evasion in vivo and form the foundation for a new approach in our efforts to develop a vaccine that prevents severe S aureusinfections
Acute and chronic transplant
rejection Therapy with protein A
Hyperacute rejection[edit]
Initiated by preexisting humoral
immunity hyperacute rejection manifests within
minutes after transplant and if tissue is left
implanted brings systemic inflammatory response
syndrome Of high risk in kidney transplants is
rapid clumping namely agglutination of red blood
cells (RBCs or erythrocytes) as an antibody
molecule binds multiple target cells at once
httpenwikipediaorgwikiOrgan_transplantation
Acute and chronic transplant
rejection Therapy with protein A
While kidneys can routinely be obtained from human donors most organs are in short supply leading to consideration of xenotransplants from other species Pigs are especially likely sources for xenotransplants chosen for the anatomical and physiological characteristics they share with humans However the sugar galactose-alpha-13-galactose(αGal) has been implicated as a major factor in hyperacute rejection in xenotransplantation Unlike virtually all other mammals humans and other primates do not make αGal and in fact recognize it as an antigen During transplantation xenoreactive natural antibodies recognize αGal on the graft endothelium as an antigen and the resulting complement-mediated immune response leads to a rejection of the transplant httpenwikipediaorgwikiOrgan_transplantation
Acute and chronic transplant
rejection Therapy with protein A
Acute rejection
Developing with formation of cellular immunity acute rejection occurs to some degree in all transplants except between identical twins unless immunosuppression is achieved (usually through drugs) Acute rejection begins as early as one week after transplant the risk being highest in the first three months though it can occur months to years later Highly vascular tissues such as kidney or liver often host the earliest signsmdashparticularly at endothelial cells lining blood vesselsmdashthough it eventually occurs in roughly 10 to 30 of liver transplants and 10 to 20 of kidney transplants httpenwikipediaorgwikiOrgan_transplantation
Acute and chronic transplant
rejection Therapy with protein A
A single episode of acute rejection can be recognized and promptly treated usually preventing organ failure but recurrent episodes lead to chronic rejection It is believed that the process of acute rejection is mediated by the cell mediated pathway specifically by mononuclear macrophages and T-lymphocytes httpenwikipediaorgwikiOrgan_transplantation
Acute and chronic transplant
rejection Therapy with protein A
Chronic Rejection
The term chronic rejection initially described long-term loss of function in transplanted organs via fibrosis of the transplanted tissues blood vessels This is now chronic allograft vasculopathy however leaving chronic rejection referring to rejection due to more patent aspects of immunity httpenwikipediaorgwikiOrgan_transplantation
Acute and chronic transplant
rejection Therapy with protein A
Chronic rejection explains long-term morbidity in most lung-transplant recipients the median survival roughly 47 years about half the span versus other major organ transplants[16] In histopathology the condition is bronchiolitisobliterans which clinically presents as progressive airflow obstruction often involving dyspnea and coughing and the patient eventually succumbs to pulmonary insufficiency or secondary acute infection
httpenwikipediaorgwikiOrgan_transplantation
Acute and chronic transplant
rejection Therapy with protein A
Airflow obstruction not ascribable to other cause is labeled bronchiolitis obliterans syndrome (BOS) confirmed by a persistent dropmdashthree or more weeksmdashin forced expiratory volume (FEV1) by at least 20[17] BOS is seen in over 50 of lung-transplant recipients by 5 years and in over 80 by ten years First noted is infiltration by lymphocytes followed by epithelial cell injury then inflammatory lesions and recruitment of fibroblasts and myofibroblasts which proliferate and secrete proteins forming scar tissue
Acute and chronic transplant
rejection Therapy with protein A
Generally thought unpredictable BOS
progression varies widely lung function may
suddenly fall but stabilize for years or rapidly
progress to death within a few months Risk
factors include prior acute rejection
episodes gastroesophageal reflux disease acute
infections particular age groups HLA mis-
matching lymphocytic bronchiolitis and graft
dysfunction (eg airway ischemia)
httpenwikipediaorgwikiOrgan_transplantation
Acute and chronic transplant
rejection Therapy with protein A
Immune Mechanisms of Transplant Rejections
Rejection is an adaptive immune response via cellular immunity (mediated by killer T cells inducing apoptosis of target cells) as well as humoral immunity (mediated by activated B cells secreting antibody molecules) though the action is joined by components of innate immune response (phagocytes and soluble immune proteins) Different types of transplanted tissues tend to favor different balances of rejection mechanisms
Acute and chronic transplant
rejection Therapy with protein A
Immunization
An animals exposure to the antigens of a different member of the same or similar species is allostimulation and the tissue is allogenic Transplanted organs are often acquired from a cadaver (usually a host who had succumbed to trauma) whose tissues had already sustained ischemia or inflammation
Dendritic cells (DCs) which are the primary antigen-presenting cells (APCs) of the donor tissue migrate to the recipients peripheral lymphoid tissue (lymphoid follicles and lymph nodes) and present the donors self peptides to the recipients lymphocytes (immune cells residing in lymphoid tissues) Lymphocytes include two classes that enact adaptive immunity also called specific immunity Lymphocytes of specific immunity T cellsmdashincluding the subclasses helper T cells and killer T cellsmdashand B cells
The recipients helper T cells coordinate specific immunity directed at the donors self peptides or at the donors Major histocompatibilitycomplex molecules or at both
Acute and chronic transplant
rejection Therapy with protein A
Immune memory
When memory helper T cells CD4 receptors bind to the MHC class II molecules which are expressed on the surfaces of the target cells of the graft tissue the memory helper T cells T cell receptors (TCRs) can recognize their target antigen that is presented by the MHC class II molecules The memory helper T cell subsequently produces clones that as effector cells secrete immune signalling molecules (cytokines) in approximately the cytokine balance that had prevailed at the memory helper T cells priming to memorize the antigen As the priming event in this instance occurred amid inflammation the immune memory is pro-inflammatory
Acute and chronic transplant
rejection Therapy with protein A
Cellular immunity
As a cell is indicated by the prefix cyto a cytotoxicinfluence destroys the cell Alloreactive killer T cells also called cytotoxic T lymphocytes (CTLs) have CD8 receptors that dock to the transplanted tissues MHC class I moleculeswhich display the donors self peptides (In the living donor such presentation of self antigens helped maintain self tolerance) Thereupon the T cell receptors (TCRs) of the killer T cells recognize their matching epitope and trigger the target cells programmed cell death by apoptosis
Acute and chronic transplant
rejection Therapy with protein A
Humoral immunity
Developed through an earlier primary exposure that primed specific immunity to the nonself antigen a transplant recipient can have specific antibody crossreacting with the donor tissue upon the transplant event a secondary exposure This is typical after earlier mismatching among ABO blood types during blood transfusion At this secondary exposure these crossreactive antibody molecules interact with aspects of innate immunitymdashsoluble immune proteins called complement and innate immune cells calledphagocytesmdashwhich inflames and destroys the transplanted tissue
Acute and chronic transplant
rejection Therapy with protein A
Antibody[edit]
Secreted by an activated B cell then
called plasma cell an antibody molecule is a
soluble immunoglobulin (Ig) whose basic unit is
shaped like the letter Y the two arms are theFab
regions while the single stalk is the Fc region
Each of the two tips of Fab region is the paratope
which binds a matching molecular sequence and
its 3D shape (conformation) altogether
called epitope within the target antigen
Acute and chronic transplant
rejection Therapy with protein A
Opsonization
The IgGs Fc region also enables opsonization by a phagocyte a process by which the Fc receptor on the phagocytemdashsuch as neutrophils in blood and macrophages in tissuesmdashbinds the antibody molecules FC stalk and the phagocyte exhibits enhanced uptake of the antigen attached to the antibody molecules Fab region
Acute and chronic transplant
rejection Therapy with protein A
Complement cascade
When the paratope of Ig class gamma (IgG) binds its matching epitope IgGs Fc region conformationally shifts and can host a complement protein initiating the complement cascade that terminates by punching a hole in a cell membrane With many holes so punched fluid rushes into the cell and ruptures it
Cell debris can be recognized as damage associated molecular patterns (DAMPs) by pattern recognition receptors (PRRs) such as Toll-like receptors (TLRs) on membranes ofphagocytes which thereupon secrete proinflammatory cytokines recruiting more phagocytes to traffic to the area by sensing the concentration gradient of the secreted cytokines (chemotaxis)
Acute and chronic transplant
rejection Therapy with protein A
Protein A is a
42 kDa surface protein originally found in the
cell wall of the bacterium Staphylococcus
aureus It is encoded by the spa gene and its
regulation is controlled by DNA topology
cellular osmolarity and a two-component
system called ArlS-ArlR It has found use in
biochemical research because of its ability to
bind immunoglobulins
Acute and chronic transplant
rejection Therapy with protein A
It is composed of five homologous Ig-binding domains that fold into a three-helix bundle Each domain is able to bind proteins from many mammalian species most notably IgGs It binds the heavy chain within the Fc region of most immunoglobulins and also within the Fab region in the case of the human VH3 family Through these interactions in serum where IgG molecules are bound in the wrong orientation (in relation to normal antibodyfunction) the bacteria disrupts opsonization and phagocytosis
Acute and chronic transplant
rejection Therapy with protein A
Purified (unconjugated) Thermo Scientific Pierce Recombinant Protein A is useful as the basis for preparing various kinds of probes or affinity media for detection or purification of rabbit and human antibodies especially IgGisotypes in immunoassays and antibody purification protocols
Features of Recombinant Protein A
bull Contains four Fc-binding domains per proteinbull Better than Protein G for polyclonal IgG from rabbit pig dog and cat serumbull Poorer than Protein G for mouse IgG1 human IgG3 and rat goat and cow antibodies
Acute and chronic transplant
rejection Therapy with protein A
Protein A is a cell wall component produced by several strains of Staphylococcus aureus that consists of a single polypeptide chain and contains little or no carbohydrate Recombinant Protein A is produced in E coli and functions essentially the same as native Protein A The Protein A molecule contains four high-affinity binding sites capable of interacting with the Fcregion from IgG of several species including human and rabbit Optimal binding occurs at pH 82 although binding is also effective at neutral or physiological conditions (pH 70 to 76)
Acute and chronic transplant
rejection Therapy with protein A
httpswwwlifetechnologiescomordercatalog
product21184
Acute and chronic transplant
rejection Therapy with protein A
httpstoolslifetechnologiescomcontentsfsm
anualsMAN0011438_Pierce_Ig_Bind_Protein
s_UGpdf
httpstoolslifetechnologiescomcontentsfsbr
ochuresTR0034-Ab-binding-proteinspdf
Acute and chronic transplant
rejection Therapy with protein A
Crystal structure of a Staphylococcus
aureus protein A domain complexed with
the Fab fragment of a human IgM antibody
Structural basis for recognition of B-cell
receptors and superantigen activity Marc Graille et al Proc Natl Acad Sci U S A
2000 May 9 97(10) 5399ndash5404 PMCID PMC25840
Immunology
Acute and chronic transplant
rejection Therapy with protein A
Staphylococcus aureus Protein A Promotes
Immune Suppression
Scott D Kobayashi
Frank R DeLeo
Acute and chronic transplant
rejection Therapy with protein A
SpA binds the Fc region of antibody and the Fabregions of the B-cell receptor processes that are known to block opsonophagocytosis and cause B-cell death in vitro In a recent study Falugi et al [F Falugi H K Kim D M Missiakas and O Schneewind mBio4(5)e00575-13 2013] showed that vaccination with spa mutant S aureus strains lacking antibody Fc-andor Fab-binding capacity protects against subsequent challenge with the USA300 epidemic strain The findings provide strong support for the idea that SpA promotes S aureus immune evasion in vivo and form the foundation for a new approach in our efforts to develop a vaccine that prevents severe S aureusinfections
Acute and chronic transplant
rejection Therapy with protein A
While kidneys can routinely be obtained from human donors most organs are in short supply leading to consideration of xenotransplants from other species Pigs are especially likely sources for xenotransplants chosen for the anatomical and physiological characteristics they share with humans However the sugar galactose-alpha-13-galactose(αGal) has been implicated as a major factor in hyperacute rejection in xenotransplantation Unlike virtually all other mammals humans and other primates do not make αGal and in fact recognize it as an antigen During transplantation xenoreactive natural antibodies recognize αGal on the graft endothelium as an antigen and the resulting complement-mediated immune response leads to a rejection of the transplant httpenwikipediaorgwikiOrgan_transplantation
Acute and chronic transplant
rejection Therapy with protein A
Acute rejection
Developing with formation of cellular immunity acute rejection occurs to some degree in all transplants except between identical twins unless immunosuppression is achieved (usually through drugs) Acute rejection begins as early as one week after transplant the risk being highest in the first three months though it can occur months to years later Highly vascular tissues such as kidney or liver often host the earliest signsmdashparticularly at endothelial cells lining blood vesselsmdashthough it eventually occurs in roughly 10 to 30 of liver transplants and 10 to 20 of kidney transplants httpenwikipediaorgwikiOrgan_transplantation
Acute and chronic transplant
rejection Therapy with protein A
A single episode of acute rejection can be recognized and promptly treated usually preventing organ failure but recurrent episodes lead to chronic rejection It is believed that the process of acute rejection is mediated by the cell mediated pathway specifically by mononuclear macrophages and T-lymphocytes httpenwikipediaorgwikiOrgan_transplantation
Acute and chronic transplant
rejection Therapy with protein A
Chronic Rejection
The term chronic rejection initially described long-term loss of function in transplanted organs via fibrosis of the transplanted tissues blood vessels This is now chronic allograft vasculopathy however leaving chronic rejection referring to rejection due to more patent aspects of immunity httpenwikipediaorgwikiOrgan_transplantation
Acute and chronic transplant
rejection Therapy with protein A
Chronic rejection explains long-term morbidity in most lung-transplant recipients the median survival roughly 47 years about half the span versus other major organ transplants[16] In histopathology the condition is bronchiolitisobliterans which clinically presents as progressive airflow obstruction often involving dyspnea and coughing and the patient eventually succumbs to pulmonary insufficiency or secondary acute infection
httpenwikipediaorgwikiOrgan_transplantation
Acute and chronic transplant
rejection Therapy with protein A
Airflow obstruction not ascribable to other cause is labeled bronchiolitis obliterans syndrome (BOS) confirmed by a persistent dropmdashthree or more weeksmdashin forced expiratory volume (FEV1) by at least 20[17] BOS is seen in over 50 of lung-transplant recipients by 5 years and in over 80 by ten years First noted is infiltration by lymphocytes followed by epithelial cell injury then inflammatory lesions and recruitment of fibroblasts and myofibroblasts which proliferate and secrete proteins forming scar tissue
Acute and chronic transplant
rejection Therapy with protein A
Generally thought unpredictable BOS
progression varies widely lung function may
suddenly fall but stabilize for years or rapidly
progress to death within a few months Risk
factors include prior acute rejection
episodes gastroesophageal reflux disease acute
infections particular age groups HLA mis-
matching lymphocytic bronchiolitis and graft
dysfunction (eg airway ischemia)
httpenwikipediaorgwikiOrgan_transplantation
Acute and chronic transplant
rejection Therapy with protein A
Immune Mechanisms of Transplant Rejections
Rejection is an adaptive immune response via cellular immunity (mediated by killer T cells inducing apoptosis of target cells) as well as humoral immunity (mediated by activated B cells secreting antibody molecules) though the action is joined by components of innate immune response (phagocytes and soluble immune proteins) Different types of transplanted tissues tend to favor different balances of rejection mechanisms
Acute and chronic transplant
rejection Therapy with protein A
Immunization
An animals exposure to the antigens of a different member of the same or similar species is allostimulation and the tissue is allogenic Transplanted organs are often acquired from a cadaver (usually a host who had succumbed to trauma) whose tissues had already sustained ischemia or inflammation
Dendritic cells (DCs) which are the primary antigen-presenting cells (APCs) of the donor tissue migrate to the recipients peripheral lymphoid tissue (lymphoid follicles and lymph nodes) and present the donors self peptides to the recipients lymphocytes (immune cells residing in lymphoid tissues) Lymphocytes include two classes that enact adaptive immunity also called specific immunity Lymphocytes of specific immunity T cellsmdashincluding the subclasses helper T cells and killer T cellsmdashand B cells
The recipients helper T cells coordinate specific immunity directed at the donors self peptides or at the donors Major histocompatibilitycomplex molecules or at both
Acute and chronic transplant
rejection Therapy with protein A
Immune memory
When memory helper T cells CD4 receptors bind to the MHC class II molecules which are expressed on the surfaces of the target cells of the graft tissue the memory helper T cells T cell receptors (TCRs) can recognize their target antigen that is presented by the MHC class II molecules The memory helper T cell subsequently produces clones that as effector cells secrete immune signalling molecules (cytokines) in approximately the cytokine balance that had prevailed at the memory helper T cells priming to memorize the antigen As the priming event in this instance occurred amid inflammation the immune memory is pro-inflammatory
Acute and chronic transplant
rejection Therapy with protein A
Cellular immunity
As a cell is indicated by the prefix cyto a cytotoxicinfluence destroys the cell Alloreactive killer T cells also called cytotoxic T lymphocytes (CTLs) have CD8 receptors that dock to the transplanted tissues MHC class I moleculeswhich display the donors self peptides (In the living donor such presentation of self antigens helped maintain self tolerance) Thereupon the T cell receptors (TCRs) of the killer T cells recognize their matching epitope and trigger the target cells programmed cell death by apoptosis
Acute and chronic transplant
rejection Therapy with protein A
Humoral immunity
Developed through an earlier primary exposure that primed specific immunity to the nonself antigen a transplant recipient can have specific antibody crossreacting with the donor tissue upon the transplant event a secondary exposure This is typical after earlier mismatching among ABO blood types during blood transfusion At this secondary exposure these crossreactive antibody molecules interact with aspects of innate immunitymdashsoluble immune proteins called complement and innate immune cells calledphagocytesmdashwhich inflames and destroys the transplanted tissue
Acute and chronic transplant
rejection Therapy with protein A
Antibody[edit]
Secreted by an activated B cell then
called plasma cell an antibody molecule is a
soluble immunoglobulin (Ig) whose basic unit is
shaped like the letter Y the two arms are theFab
regions while the single stalk is the Fc region
Each of the two tips of Fab region is the paratope
which binds a matching molecular sequence and
its 3D shape (conformation) altogether
called epitope within the target antigen
Acute and chronic transplant
rejection Therapy with protein A
Opsonization
The IgGs Fc region also enables opsonization by a phagocyte a process by which the Fc receptor on the phagocytemdashsuch as neutrophils in blood and macrophages in tissuesmdashbinds the antibody molecules FC stalk and the phagocyte exhibits enhanced uptake of the antigen attached to the antibody molecules Fab region
Acute and chronic transplant
rejection Therapy with protein A
Complement cascade
When the paratope of Ig class gamma (IgG) binds its matching epitope IgGs Fc region conformationally shifts and can host a complement protein initiating the complement cascade that terminates by punching a hole in a cell membrane With many holes so punched fluid rushes into the cell and ruptures it
Cell debris can be recognized as damage associated molecular patterns (DAMPs) by pattern recognition receptors (PRRs) such as Toll-like receptors (TLRs) on membranes ofphagocytes which thereupon secrete proinflammatory cytokines recruiting more phagocytes to traffic to the area by sensing the concentration gradient of the secreted cytokines (chemotaxis)
Acute and chronic transplant
rejection Therapy with protein A
Protein A is a
42 kDa surface protein originally found in the
cell wall of the bacterium Staphylococcus
aureus It is encoded by the spa gene and its
regulation is controlled by DNA topology
cellular osmolarity and a two-component
system called ArlS-ArlR It has found use in
biochemical research because of its ability to
bind immunoglobulins
Acute and chronic transplant
rejection Therapy with protein A
It is composed of five homologous Ig-binding domains that fold into a three-helix bundle Each domain is able to bind proteins from many mammalian species most notably IgGs It binds the heavy chain within the Fc region of most immunoglobulins and also within the Fab region in the case of the human VH3 family Through these interactions in serum where IgG molecules are bound in the wrong orientation (in relation to normal antibodyfunction) the bacteria disrupts opsonization and phagocytosis
Acute and chronic transplant
rejection Therapy with protein A
Purified (unconjugated) Thermo Scientific Pierce Recombinant Protein A is useful as the basis for preparing various kinds of probes or affinity media for detection or purification of rabbit and human antibodies especially IgGisotypes in immunoassays and antibody purification protocols
Features of Recombinant Protein A
bull Contains four Fc-binding domains per proteinbull Better than Protein G for polyclonal IgG from rabbit pig dog and cat serumbull Poorer than Protein G for mouse IgG1 human IgG3 and rat goat and cow antibodies
Acute and chronic transplant
rejection Therapy with protein A
Protein A is a cell wall component produced by several strains of Staphylococcus aureus that consists of a single polypeptide chain and contains little or no carbohydrate Recombinant Protein A is produced in E coli and functions essentially the same as native Protein A The Protein A molecule contains four high-affinity binding sites capable of interacting with the Fcregion from IgG of several species including human and rabbit Optimal binding occurs at pH 82 although binding is also effective at neutral or physiological conditions (pH 70 to 76)
Acute and chronic transplant
rejection Therapy with protein A
httpswwwlifetechnologiescomordercatalog
product21184
Acute and chronic transplant
rejection Therapy with protein A
httpstoolslifetechnologiescomcontentsfsm
anualsMAN0011438_Pierce_Ig_Bind_Protein
s_UGpdf
httpstoolslifetechnologiescomcontentsfsbr
ochuresTR0034-Ab-binding-proteinspdf
Acute and chronic transplant
rejection Therapy with protein A
Crystal structure of a Staphylococcus
aureus protein A domain complexed with
the Fab fragment of a human IgM antibody
Structural basis for recognition of B-cell
receptors and superantigen activity Marc Graille et al Proc Natl Acad Sci U S A
2000 May 9 97(10) 5399ndash5404 PMCID PMC25840
Immunology
Acute and chronic transplant
rejection Therapy with protein A
Staphylococcus aureus Protein A Promotes
Immune Suppression
Scott D Kobayashi
Frank R DeLeo
Acute and chronic transplant
rejection Therapy with protein A
SpA binds the Fc region of antibody and the Fabregions of the B-cell receptor processes that are known to block opsonophagocytosis and cause B-cell death in vitro In a recent study Falugi et al [F Falugi H K Kim D M Missiakas and O Schneewind mBio4(5)e00575-13 2013] showed that vaccination with spa mutant S aureus strains lacking antibody Fc-andor Fab-binding capacity protects against subsequent challenge with the USA300 epidemic strain The findings provide strong support for the idea that SpA promotes S aureus immune evasion in vivo and form the foundation for a new approach in our efforts to develop a vaccine that prevents severe S aureusinfections
Acute and chronic transplant
rejection Therapy with protein A
Acute rejection
Developing with formation of cellular immunity acute rejection occurs to some degree in all transplants except between identical twins unless immunosuppression is achieved (usually through drugs) Acute rejection begins as early as one week after transplant the risk being highest in the first three months though it can occur months to years later Highly vascular tissues such as kidney or liver often host the earliest signsmdashparticularly at endothelial cells lining blood vesselsmdashthough it eventually occurs in roughly 10 to 30 of liver transplants and 10 to 20 of kidney transplants httpenwikipediaorgwikiOrgan_transplantation
Acute and chronic transplant
rejection Therapy with protein A
A single episode of acute rejection can be recognized and promptly treated usually preventing organ failure but recurrent episodes lead to chronic rejection It is believed that the process of acute rejection is mediated by the cell mediated pathway specifically by mononuclear macrophages and T-lymphocytes httpenwikipediaorgwikiOrgan_transplantation
Acute and chronic transplant
rejection Therapy with protein A
Chronic Rejection
The term chronic rejection initially described long-term loss of function in transplanted organs via fibrosis of the transplanted tissues blood vessels This is now chronic allograft vasculopathy however leaving chronic rejection referring to rejection due to more patent aspects of immunity httpenwikipediaorgwikiOrgan_transplantation
Acute and chronic transplant
rejection Therapy with protein A
Chronic rejection explains long-term morbidity in most lung-transplant recipients the median survival roughly 47 years about half the span versus other major organ transplants[16] In histopathology the condition is bronchiolitisobliterans which clinically presents as progressive airflow obstruction often involving dyspnea and coughing and the patient eventually succumbs to pulmonary insufficiency or secondary acute infection
httpenwikipediaorgwikiOrgan_transplantation
Acute and chronic transplant
rejection Therapy with protein A
Airflow obstruction not ascribable to other cause is labeled bronchiolitis obliterans syndrome (BOS) confirmed by a persistent dropmdashthree or more weeksmdashin forced expiratory volume (FEV1) by at least 20[17] BOS is seen in over 50 of lung-transplant recipients by 5 years and in over 80 by ten years First noted is infiltration by lymphocytes followed by epithelial cell injury then inflammatory lesions and recruitment of fibroblasts and myofibroblasts which proliferate and secrete proteins forming scar tissue
Acute and chronic transplant
rejection Therapy with protein A
Generally thought unpredictable BOS
progression varies widely lung function may
suddenly fall but stabilize for years or rapidly
progress to death within a few months Risk
factors include prior acute rejection
episodes gastroesophageal reflux disease acute
infections particular age groups HLA mis-
matching lymphocytic bronchiolitis and graft
dysfunction (eg airway ischemia)
httpenwikipediaorgwikiOrgan_transplantation
Acute and chronic transplant
rejection Therapy with protein A
Immune Mechanisms of Transplant Rejections
Rejection is an adaptive immune response via cellular immunity (mediated by killer T cells inducing apoptosis of target cells) as well as humoral immunity (mediated by activated B cells secreting antibody molecules) though the action is joined by components of innate immune response (phagocytes and soluble immune proteins) Different types of transplanted tissues tend to favor different balances of rejection mechanisms
Acute and chronic transplant
rejection Therapy with protein A
Immunization
An animals exposure to the antigens of a different member of the same or similar species is allostimulation and the tissue is allogenic Transplanted organs are often acquired from a cadaver (usually a host who had succumbed to trauma) whose tissues had already sustained ischemia or inflammation
Dendritic cells (DCs) which are the primary antigen-presenting cells (APCs) of the donor tissue migrate to the recipients peripheral lymphoid tissue (lymphoid follicles and lymph nodes) and present the donors self peptides to the recipients lymphocytes (immune cells residing in lymphoid tissues) Lymphocytes include two classes that enact adaptive immunity also called specific immunity Lymphocytes of specific immunity T cellsmdashincluding the subclasses helper T cells and killer T cellsmdashand B cells
The recipients helper T cells coordinate specific immunity directed at the donors self peptides or at the donors Major histocompatibilitycomplex molecules or at both
Acute and chronic transplant
rejection Therapy with protein A
Immune memory
When memory helper T cells CD4 receptors bind to the MHC class II molecules which are expressed on the surfaces of the target cells of the graft tissue the memory helper T cells T cell receptors (TCRs) can recognize their target antigen that is presented by the MHC class II molecules The memory helper T cell subsequently produces clones that as effector cells secrete immune signalling molecules (cytokines) in approximately the cytokine balance that had prevailed at the memory helper T cells priming to memorize the antigen As the priming event in this instance occurred amid inflammation the immune memory is pro-inflammatory
Acute and chronic transplant
rejection Therapy with protein A
Cellular immunity
As a cell is indicated by the prefix cyto a cytotoxicinfluence destroys the cell Alloreactive killer T cells also called cytotoxic T lymphocytes (CTLs) have CD8 receptors that dock to the transplanted tissues MHC class I moleculeswhich display the donors self peptides (In the living donor such presentation of self antigens helped maintain self tolerance) Thereupon the T cell receptors (TCRs) of the killer T cells recognize their matching epitope and trigger the target cells programmed cell death by apoptosis
Acute and chronic transplant
rejection Therapy with protein A
Humoral immunity
Developed through an earlier primary exposure that primed specific immunity to the nonself antigen a transplant recipient can have specific antibody crossreacting with the donor tissue upon the transplant event a secondary exposure This is typical after earlier mismatching among ABO blood types during blood transfusion At this secondary exposure these crossreactive antibody molecules interact with aspects of innate immunitymdashsoluble immune proteins called complement and innate immune cells calledphagocytesmdashwhich inflames and destroys the transplanted tissue
Acute and chronic transplant
rejection Therapy with protein A
Antibody[edit]
Secreted by an activated B cell then
called plasma cell an antibody molecule is a
soluble immunoglobulin (Ig) whose basic unit is
shaped like the letter Y the two arms are theFab
regions while the single stalk is the Fc region
Each of the two tips of Fab region is the paratope
which binds a matching molecular sequence and
its 3D shape (conformation) altogether
called epitope within the target antigen
Acute and chronic transplant
rejection Therapy with protein A
Opsonization
The IgGs Fc region also enables opsonization by a phagocyte a process by which the Fc receptor on the phagocytemdashsuch as neutrophils in blood and macrophages in tissuesmdashbinds the antibody molecules FC stalk and the phagocyte exhibits enhanced uptake of the antigen attached to the antibody molecules Fab region
Acute and chronic transplant
rejection Therapy with protein A
Complement cascade
When the paratope of Ig class gamma (IgG) binds its matching epitope IgGs Fc region conformationally shifts and can host a complement protein initiating the complement cascade that terminates by punching a hole in a cell membrane With many holes so punched fluid rushes into the cell and ruptures it
Cell debris can be recognized as damage associated molecular patterns (DAMPs) by pattern recognition receptors (PRRs) such as Toll-like receptors (TLRs) on membranes ofphagocytes which thereupon secrete proinflammatory cytokines recruiting more phagocytes to traffic to the area by sensing the concentration gradient of the secreted cytokines (chemotaxis)
Acute and chronic transplant
rejection Therapy with protein A
Protein A is a
42 kDa surface protein originally found in the
cell wall of the bacterium Staphylococcus
aureus It is encoded by the spa gene and its
regulation is controlled by DNA topology
cellular osmolarity and a two-component
system called ArlS-ArlR It has found use in
biochemical research because of its ability to
bind immunoglobulins
Acute and chronic transplant
rejection Therapy with protein A
It is composed of five homologous Ig-binding domains that fold into a three-helix bundle Each domain is able to bind proteins from many mammalian species most notably IgGs It binds the heavy chain within the Fc region of most immunoglobulins and also within the Fab region in the case of the human VH3 family Through these interactions in serum where IgG molecules are bound in the wrong orientation (in relation to normal antibodyfunction) the bacteria disrupts opsonization and phagocytosis
Acute and chronic transplant
rejection Therapy with protein A
Purified (unconjugated) Thermo Scientific Pierce Recombinant Protein A is useful as the basis for preparing various kinds of probes or affinity media for detection or purification of rabbit and human antibodies especially IgGisotypes in immunoassays and antibody purification protocols
Features of Recombinant Protein A
bull Contains four Fc-binding domains per proteinbull Better than Protein G for polyclonal IgG from rabbit pig dog and cat serumbull Poorer than Protein G for mouse IgG1 human IgG3 and rat goat and cow antibodies
Acute and chronic transplant
rejection Therapy with protein A
Protein A is a cell wall component produced by several strains of Staphylococcus aureus that consists of a single polypeptide chain and contains little or no carbohydrate Recombinant Protein A is produced in E coli and functions essentially the same as native Protein A The Protein A molecule contains four high-affinity binding sites capable of interacting with the Fcregion from IgG of several species including human and rabbit Optimal binding occurs at pH 82 although binding is also effective at neutral or physiological conditions (pH 70 to 76)
Acute and chronic transplant
rejection Therapy with protein A
httpswwwlifetechnologiescomordercatalog
product21184
Acute and chronic transplant
rejection Therapy with protein A
httpstoolslifetechnologiescomcontentsfsm
anualsMAN0011438_Pierce_Ig_Bind_Protein
s_UGpdf
httpstoolslifetechnologiescomcontentsfsbr
ochuresTR0034-Ab-binding-proteinspdf
Acute and chronic transplant
rejection Therapy with protein A
Crystal structure of a Staphylococcus
aureus protein A domain complexed with
the Fab fragment of a human IgM antibody
Structural basis for recognition of B-cell
receptors and superantigen activity Marc Graille et al Proc Natl Acad Sci U S A
2000 May 9 97(10) 5399ndash5404 PMCID PMC25840
Immunology
Acute and chronic transplant
rejection Therapy with protein A
Staphylococcus aureus Protein A Promotes
Immune Suppression
Scott D Kobayashi
Frank R DeLeo
Acute and chronic transplant
rejection Therapy with protein A
SpA binds the Fc region of antibody and the Fabregions of the B-cell receptor processes that are known to block opsonophagocytosis and cause B-cell death in vitro In a recent study Falugi et al [F Falugi H K Kim D M Missiakas and O Schneewind mBio4(5)e00575-13 2013] showed that vaccination with spa mutant S aureus strains lacking antibody Fc-andor Fab-binding capacity protects against subsequent challenge with the USA300 epidemic strain The findings provide strong support for the idea that SpA promotes S aureus immune evasion in vivo and form the foundation for a new approach in our efforts to develop a vaccine that prevents severe S aureusinfections
Acute and chronic transplant
rejection Therapy with protein A
A single episode of acute rejection can be recognized and promptly treated usually preventing organ failure but recurrent episodes lead to chronic rejection It is believed that the process of acute rejection is mediated by the cell mediated pathway specifically by mononuclear macrophages and T-lymphocytes httpenwikipediaorgwikiOrgan_transplantation
Acute and chronic transplant
rejection Therapy with protein A
Chronic Rejection
The term chronic rejection initially described long-term loss of function in transplanted organs via fibrosis of the transplanted tissues blood vessels This is now chronic allograft vasculopathy however leaving chronic rejection referring to rejection due to more patent aspects of immunity httpenwikipediaorgwikiOrgan_transplantation
Acute and chronic transplant
rejection Therapy with protein A
Chronic rejection explains long-term morbidity in most lung-transplant recipients the median survival roughly 47 years about half the span versus other major organ transplants[16] In histopathology the condition is bronchiolitisobliterans which clinically presents as progressive airflow obstruction often involving dyspnea and coughing and the patient eventually succumbs to pulmonary insufficiency or secondary acute infection
httpenwikipediaorgwikiOrgan_transplantation
Acute and chronic transplant
rejection Therapy with protein A
Airflow obstruction not ascribable to other cause is labeled bronchiolitis obliterans syndrome (BOS) confirmed by a persistent dropmdashthree or more weeksmdashin forced expiratory volume (FEV1) by at least 20[17] BOS is seen in over 50 of lung-transplant recipients by 5 years and in over 80 by ten years First noted is infiltration by lymphocytes followed by epithelial cell injury then inflammatory lesions and recruitment of fibroblasts and myofibroblasts which proliferate and secrete proteins forming scar tissue
Acute and chronic transplant
rejection Therapy with protein A
Generally thought unpredictable BOS
progression varies widely lung function may
suddenly fall but stabilize for years or rapidly
progress to death within a few months Risk
factors include prior acute rejection
episodes gastroesophageal reflux disease acute
infections particular age groups HLA mis-
matching lymphocytic bronchiolitis and graft
dysfunction (eg airway ischemia)
httpenwikipediaorgwikiOrgan_transplantation
Acute and chronic transplant
rejection Therapy with protein A
Immune Mechanisms of Transplant Rejections
Rejection is an adaptive immune response via cellular immunity (mediated by killer T cells inducing apoptosis of target cells) as well as humoral immunity (mediated by activated B cells secreting antibody molecules) though the action is joined by components of innate immune response (phagocytes and soluble immune proteins) Different types of transplanted tissues tend to favor different balances of rejection mechanisms
Acute and chronic transplant
rejection Therapy with protein A
Immunization
An animals exposure to the antigens of a different member of the same or similar species is allostimulation and the tissue is allogenic Transplanted organs are often acquired from a cadaver (usually a host who had succumbed to trauma) whose tissues had already sustained ischemia or inflammation
Dendritic cells (DCs) which are the primary antigen-presenting cells (APCs) of the donor tissue migrate to the recipients peripheral lymphoid tissue (lymphoid follicles and lymph nodes) and present the donors self peptides to the recipients lymphocytes (immune cells residing in lymphoid tissues) Lymphocytes include two classes that enact adaptive immunity also called specific immunity Lymphocytes of specific immunity T cellsmdashincluding the subclasses helper T cells and killer T cellsmdashand B cells
The recipients helper T cells coordinate specific immunity directed at the donors self peptides or at the donors Major histocompatibilitycomplex molecules or at both
Acute and chronic transplant
rejection Therapy with protein A
Immune memory
When memory helper T cells CD4 receptors bind to the MHC class II molecules which are expressed on the surfaces of the target cells of the graft tissue the memory helper T cells T cell receptors (TCRs) can recognize their target antigen that is presented by the MHC class II molecules The memory helper T cell subsequently produces clones that as effector cells secrete immune signalling molecules (cytokines) in approximately the cytokine balance that had prevailed at the memory helper T cells priming to memorize the antigen As the priming event in this instance occurred amid inflammation the immune memory is pro-inflammatory
Acute and chronic transplant
rejection Therapy with protein A
Cellular immunity
As a cell is indicated by the prefix cyto a cytotoxicinfluence destroys the cell Alloreactive killer T cells also called cytotoxic T lymphocytes (CTLs) have CD8 receptors that dock to the transplanted tissues MHC class I moleculeswhich display the donors self peptides (In the living donor such presentation of self antigens helped maintain self tolerance) Thereupon the T cell receptors (TCRs) of the killer T cells recognize their matching epitope and trigger the target cells programmed cell death by apoptosis
Acute and chronic transplant
rejection Therapy with protein A
Humoral immunity
Developed through an earlier primary exposure that primed specific immunity to the nonself antigen a transplant recipient can have specific antibody crossreacting with the donor tissue upon the transplant event a secondary exposure This is typical after earlier mismatching among ABO blood types during blood transfusion At this secondary exposure these crossreactive antibody molecules interact with aspects of innate immunitymdashsoluble immune proteins called complement and innate immune cells calledphagocytesmdashwhich inflames and destroys the transplanted tissue
Acute and chronic transplant
rejection Therapy with protein A
Antibody[edit]
Secreted by an activated B cell then
called plasma cell an antibody molecule is a
soluble immunoglobulin (Ig) whose basic unit is
shaped like the letter Y the two arms are theFab
regions while the single stalk is the Fc region
Each of the two tips of Fab region is the paratope
which binds a matching molecular sequence and
its 3D shape (conformation) altogether
called epitope within the target antigen
Acute and chronic transplant
rejection Therapy with protein A
Opsonization
The IgGs Fc region also enables opsonization by a phagocyte a process by which the Fc receptor on the phagocytemdashsuch as neutrophils in blood and macrophages in tissuesmdashbinds the antibody molecules FC stalk and the phagocyte exhibits enhanced uptake of the antigen attached to the antibody molecules Fab region
Acute and chronic transplant
rejection Therapy with protein A
Complement cascade
When the paratope of Ig class gamma (IgG) binds its matching epitope IgGs Fc region conformationally shifts and can host a complement protein initiating the complement cascade that terminates by punching a hole in a cell membrane With many holes so punched fluid rushes into the cell and ruptures it
Cell debris can be recognized as damage associated molecular patterns (DAMPs) by pattern recognition receptors (PRRs) such as Toll-like receptors (TLRs) on membranes ofphagocytes which thereupon secrete proinflammatory cytokines recruiting more phagocytes to traffic to the area by sensing the concentration gradient of the secreted cytokines (chemotaxis)
Acute and chronic transplant
rejection Therapy with protein A
Protein A is a
42 kDa surface protein originally found in the
cell wall of the bacterium Staphylococcus
aureus It is encoded by the spa gene and its
regulation is controlled by DNA topology
cellular osmolarity and a two-component
system called ArlS-ArlR It has found use in
biochemical research because of its ability to
bind immunoglobulins
Acute and chronic transplant
rejection Therapy with protein A
It is composed of five homologous Ig-binding domains that fold into a three-helix bundle Each domain is able to bind proteins from many mammalian species most notably IgGs It binds the heavy chain within the Fc region of most immunoglobulins and also within the Fab region in the case of the human VH3 family Through these interactions in serum where IgG molecules are bound in the wrong orientation (in relation to normal antibodyfunction) the bacteria disrupts opsonization and phagocytosis
Acute and chronic transplant
rejection Therapy with protein A
Purified (unconjugated) Thermo Scientific Pierce Recombinant Protein A is useful as the basis for preparing various kinds of probes or affinity media for detection or purification of rabbit and human antibodies especially IgGisotypes in immunoassays and antibody purification protocols
Features of Recombinant Protein A
bull Contains four Fc-binding domains per proteinbull Better than Protein G for polyclonal IgG from rabbit pig dog and cat serumbull Poorer than Protein G for mouse IgG1 human IgG3 and rat goat and cow antibodies
Acute and chronic transplant
rejection Therapy with protein A
Protein A is a cell wall component produced by several strains of Staphylococcus aureus that consists of a single polypeptide chain and contains little or no carbohydrate Recombinant Protein A is produced in E coli and functions essentially the same as native Protein A The Protein A molecule contains four high-affinity binding sites capable of interacting with the Fcregion from IgG of several species including human and rabbit Optimal binding occurs at pH 82 although binding is also effective at neutral or physiological conditions (pH 70 to 76)
Acute and chronic transplant
rejection Therapy with protein A
httpswwwlifetechnologiescomordercatalog
product21184
Acute and chronic transplant
rejection Therapy with protein A
httpstoolslifetechnologiescomcontentsfsm
anualsMAN0011438_Pierce_Ig_Bind_Protein
s_UGpdf
httpstoolslifetechnologiescomcontentsfsbr
ochuresTR0034-Ab-binding-proteinspdf
Acute and chronic transplant
rejection Therapy with protein A
Crystal structure of a Staphylococcus
aureus protein A domain complexed with
the Fab fragment of a human IgM antibody
Structural basis for recognition of B-cell
receptors and superantigen activity Marc Graille et al Proc Natl Acad Sci U S A
2000 May 9 97(10) 5399ndash5404 PMCID PMC25840
Immunology
Acute and chronic transplant
rejection Therapy with protein A
Staphylococcus aureus Protein A Promotes
Immune Suppression
Scott D Kobayashi
Frank R DeLeo
Acute and chronic transplant
rejection Therapy with protein A
SpA binds the Fc region of antibody and the Fabregions of the B-cell receptor processes that are known to block opsonophagocytosis and cause B-cell death in vitro In a recent study Falugi et al [F Falugi H K Kim D M Missiakas and O Schneewind mBio4(5)e00575-13 2013] showed that vaccination with spa mutant S aureus strains lacking antibody Fc-andor Fab-binding capacity protects against subsequent challenge with the USA300 epidemic strain The findings provide strong support for the idea that SpA promotes S aureus immune evasion in vivo and form the foundation for a new approach in our efforts to develop a vaccine that prevents severe S aureusinfections
Acute and chronic transplant
rejection Therapy with protein A
Chronic Rejection
The term chronic rejection initially described long-term loss of function in transplanted organs via fibrosis of the transplanted tissues blood vessels This is now chronic allograft vasculopathy however leaving chronic rejection referring to rejection due to more patent aspects of immunity httpenwikipediaorgwikiOrgan_transplantation
Acute and chronic transplant
rejection Therapy with protein A
Chronic rejection explains long-term morbidity in most lung-transplant recipients the median survival roughly 47 years about half the span versus other major organ transplants[16] In histopathology the condition is bronchiolitisobliterans which clinically presents as progressive airflow obstruction often involving dyspnea and coughing and the patient eventually succumbs to pulmonary insufficiency or secondary acute infection
httpenwikipediaorgwikiOrgan_transplantation
Acute and chronic transplant
rejection Therapy with protein A
Airflow obstruction not ascribable to other cause is labeled bronchiolitis obliterans syndrome (BOS) confirmed by a persistent dropmdashthree or more weeksmdashin forced expiratory volume (FEV1) by at least 20[17] BOS is seen in over 50 of lung-transplant recipients by 5 years and in over 80 by ten years First noted is infiltration by lymphocytes followed by epithelial cell injury then inflammatory lesions and recruitment of fibroblasts and myofibroblasts which proliferate and secrete proteins forming scar tissue
Acute and chronic transplant
rejection Therapy with protein A
Generally thought unpredictable BOS
progression varies widely lung function may
suddenly fall but stabilize for years or rapidly
progress to death within a few months Risk
factors include prior acute rejection
episodes gastroesophageal reflux disease acute
infections particular age groups HLA mis-
matching lymphocytic bronchiolitis and graft
dysfunction (eg airway ischemia)
httpenwikipediaorgwikiOrgan_transplantation
Acute and chronic transplant
rejection Therapy with protein A
Immune Mechanisms of Transplant Rejections
Rejection is an adaptive immune response via cellular immunity (mediated by killer T cells inducing apoptosis of target cells) as well as humoral immunity (mediated by activated B cells secreting antibody molecules) though the action is joined by components of innate immune response (phagocytes and soluble immune proteins) Different types of transplanted tissues tend to favor different balances of rejection mechanisms
Acute and chronic transplant
rejection Therapy with protein A
Immunization
An animals exposure to the antigens of a different member of the same or similar species is allostimulation and the tissue is allogenic Transplanted organs are often acquired from a cadaver (usually a host who had succumbed to trauma) whose tissues had already sustained ischemia or inflammation
Dendritic cells (DCs) which are the primary antigen-presenting cells (APCs) of the donor tissue migrate to the recipients peripheral lymphoid tissue (lymphoid follicles and lymph nodes) and present the donors self peptides to the recipients lymphocytes (immune cells residing in lymphoid tissues) Lymphocytes include two classes that enact adaptive immunity also called specific immunity Lymphocytes of specific immunity T cellsmdashincluding the subclasses helper T cells and killer T cellsmdashand B cells
The recipients helper T cells coordinate specific immunity directed at the donors self peptides or at the donors Major histocompatibilitycomplex molecules or at both
Acute and chronic transplant
rejection Therapy with protein A
Immune memory
When memory helper T cells CD4 receptors bind to the MHC class II molecules which are expressed on the surfaces of the target cells of the graft tissue the memory helper T cells T cell receptors (TCRs) can recognize their target antigen that is presented by the MHC class II molecules The memory helper T cell subsequently produces clones that as effector cells secrete immune signalling molecules (cytokines) in approximately the cytokine balance that had prevailed at the memory helper T cells priming to memorize the antigen As the priming event in this instance occurred amid inflammation the immune memory is pro-inflammatory
Acute and chronic transplant
rejection Therapy with protein A
Cellular immunity
As a cell is indicated by the prefix cyto a cytotoxicinfluence destroys the cell Alloreactive killer T cells also called cytotoxic T lymphocytes (CTLs) have CD8 receptors that dock to the transplanted tissues MHC class I moleculeswhich display the donors self peptides (In the living donor such presentation of self antigens helped maintain self tolerance) Thereupon the T cell receptors (TCRs) of the killer T cells recognize their matching epitope and trigger the target cells programmed cell death by apoptosis
Acute and chronic transplant
rejection Therapy with protein A
Humoral immunity
Developed through an earlier primary exposure that primed specific immunity to the nonself antigen a transplant recipient can have specific antibody crossreacting with the donor tissue upon the transplant event a secondary exposure This is typical after earlier mismatching among ABO blood types during blood transfusion At this secondary exposure these crossreactive antibody molecules interact with aspects of innate immunitymdashsoluble immune proteins called complement and innate immune cells calledphagocytesmdashwhich inflames and destroys the transplanted tissue
Acute and chronic transplant
rejection Therapy with protein A
Antibody[edit]
Secreted by an activated B cell then
called plasma cell an antibody molecule is a
soluble immunoglobulin (Ig) whose basic unit is
shaped like the letter Y the two arms are theFab
regions while the single stalk is the Fc region
Each of the two tips of Fab region is the paratope
which binds a matching molecular sequence and
its 3D shape (conformation) altogether
called epitope within the target antigen
Acute and chronic transplant
rejection Therapy with protein A
Opsonization
The IgGs Fc region also enables opsonization by a phagocyte a process by which the Fc receptor on the phagocytemdashsuch as neutrophils in blood and macrophages in tissuesmdashbinds the antibody molecules FC stalk and the phagocyte exhibits enhanced uptake of the antigen attached to the antibody molecules Fab region
Acute and chronic transplant
rejection Therapy with protein A
Complement cascade
When the paratope of Ig class gamma (IgG) binds its matching epitope IgGs Fc region conformationally shifts and can host a complement protein initiating the complement cascade that terminates by punching a hole in a cell membrane With many holes so punched fluid rushes into the cell and ruptures it
Cell debris can be recognized as damage associated molecular patterns (DAMPs) by pattern recognition receptors (PRRs) such as Toll-like receptors (TLRs) on membranes ofphagocytes which thereupon secrete proinflammatory cytokines recruiting more phagocytes to traffic to the area by sensing the concentration gradient of the secreted cytokines (chemotaxis)
Acute and chronic transplant
rejection Therapy with protein A
Protein A is a
42 kDa surface protein originally found in the
cell wall of the bacterium Staphylococcus
aureus It is encoded by the spa gene and its
regulation is controlled by DNA topology
cellular osmolarity and a two-component
system called ArlS-ArlR It has found use in
biochemical research because of its ability to
bind immunoglobulins
Acute and chronic transplant
rejection Therapy with protein A
It is composed of five homologous Ig-binding domains that fold into a three-helix bundle Each domain is able to bind proteins from many mammalian species most notably IgGs It binds the heavy chain within the Fc region of most immunoglobulins and also within the Fab region in the case of the human VH3 family Through these interactions in serum where IgG molecules are bound in the wrong orientation (in relation to normal antibodyfunction) the bacteria disrupts opsonization and phagocytosis
Acute and chronic transplant
rejection Therapy with protein A
Purified (unconjugated) Thermo Scientific Pierce Recombinant Protein A is useful as the basis for preparing various kinds of probes or affinity media for detection or purification of rabbit and human antibodies especially IgGisotypes in immunoassays and antibody purification protocols
Features of Recombinant Protein A
bull Contains four Fc-binding domains per proteinbull Better than Protein G for polyclonal IgG from rabbit pig dog and cat serumbull Poorer than Protein G for mouse IgG1 human IgG3 and rat goat and cow antibodies
Acute and chronic transplant
rejection Therapy with protein A
Protein A is a cell wall component produced by several strains of Staphylococcus aureus that consists of a single polypeptide chain and contains little or no carbohydrate Recombinant Protein A is produced in E coli and functions essentially the same as native Protein A The Protein A molecule contains four high-affinity binding sites capable of interacting with the Fcregion from IgG of several species including human and rabbit Optimal binding occurs at pH 82 although binding is also effective at neutral or physiological conditions (pH 70 to 76)
Acute and chronic transplant
rejection Therapy with protein A
httpswwwlifetechnologiescomordercatalog
product21184
Acute and chronic transplant
rejection Therapy with protein A
httpstoolslifetechnologiescomcontentsfsm
anualsMAN0011438_Pierce_Ig_Bind_Protein
s_UGpdf
httpstoolslifetechnologiescomcontentsfsbr
ochuresTR0034-Ab-binding-proteinspdf
Acute and chronic transplant
rejection Therapy with protein A
Crystal structure of a Staphylococcus
aureus protein A domain complexed with
the Fab fragment of a human IgM antibody
Structural basis for recognition of B-cell
receptors and superantigen activity Marc Graille et al Proc Natl Acad Sci U S A
2000 May 9 97(10) 5399ndash5404 PMCID PMC25840
Immunology
Acute and chronic transplant
rejection Therapy with protein A
Staphylococcus aureus Protein A Promotes
Immune Suppression
Scott D Kobayashi
Frank R DeLeo
Acute and chronic transplant
rejection Therapy with protein A
SpA binds the Fc region of antibody and the Fabregions of the B-cell receptor processes that are known to block opsonophagocytosis and cause B-cell death in vitro In a recent study Falugi et al [F Falugi H K Kim D M Missiakas and O Schneewind mBio4(5)e00575-13 2013] showed that vaccination with spa mutant S aureus strains lacking antibody Fc-andor Fab-binding capacity protects against subsequent challenge with the USA300 epidemic strain The findings provide strong support for the idea that SpA promotes S aureus immune evasion in vivo and form the foundation for a new approach in our efforts to develop a vaccine that prevents severe S aureusinfections
Acute and chronic transplant
rejection Therapy with protein A
Chronic rejection explains long-term morbidity in most lung-transplant recipients the median survival roughly 47 years about half the span versus other major organ transplants[16] In histopathology the condition is bronchiolitisobliterans which clinically presents as progressive airflow obstruction often involving dyspnea and coughing and the patient eventually succumbs to pulmonary insufficiency or secondary acute infection
httpenwikipediaorgwikiOrgan_transplantation
Acute and chronic transplant
rejection Therapy with protein A
Airflow obstruction not ascribable to other cause is labeled bronchiolitis obliterans syndrome (BOS) confirmed by a persistent dropmdashthree or more weeksmdashin forced expiratory volume (FEV1) by at least 20[17] BOS is seen in over 50 of lung-transplant recipients by 5 years and in over 80 by ten years First noted is infiltration by lymphocytes followed by epithelial cell injury then inflammatory lesions and recruitment of fibroblasts and myofibroblasts which proliferate and secrete proteins forming scar tissue
Acute and chronic transplant
rejection Therapy with protein A
Generally thought unpredictable BOS
progression varies widely lung function may
suddenly fall but stabilize for years or rapidly
progress to death within a few months Risk
factors include prior acute rejection
episodes gastroesophageal reflux disease acute
infections particular age groups HLA mis-
matching lymphocytic bronchiolitis and graft
dysfunction (eg airway ischemia)
httpenwikipediaorgwikiOrgan_transplantation
Acute and chronic transplant
rejection Therapy with protein A
Immune Mechanisms of Transplant Rejections
Rejection is an adaptive immune response via cellular immunity (mediated by killer T cells inducing apoptosis of target cells) as well as humoral immunity (mediated by activated B cells secreting antibody molecules) though the action is joined by components of innate immune response (phagocytes and soluble immune proteins) Different types of transplanted tissues tend to favor different balances of rejection mechanisms
Acute and chronic transplant
rejection Therapy with protein A
Immunization
An animals exposure to the antigens of a different member of the same or similar species is allostimulation and the tissue is allogenic Transplanted organs are often acquired from a cadaver (usually a host who had succumbed to trauma) whose tissues had already sustained ischemia or inflammation
Dendritic cells (DCs) which are the primary antigen-presenting cells (APCs) of the donor tissue migrate to the recipients peripheral lymphoid tissue (lymphoid follicles and lymph nodes) and present the donors self peptides to the recipients lymphocytes (immune cells residing in lymphoid tissues) Lymphocytes include two classes that enact adaptive immunity also called specific immunity Lymphocytes of specific immunity T cellsmdashincluding the subclasses helper T cells and killer T cellsmdashand B cells
The recipients helper T cells coordinate specific immunity directed at the donors self peptides or at the donors Major histocompatibilitycomplex molecules or at both
Acute and chronic transplant
rejection Therapy with protein A
Immune memory
When memory helper T cells CD4 receptors bind to the MHC class II molecules which are expressed on the surfaces of the target cells of the graft tissue the memory helper T cells T cell receptors (TCRs) can recognize their target antigen that is presented by the MHC class II molecules The memory helper T cell subsequently produces clones that as effector cells secrete immune signalling molecules (cytokines) in approximately the cytokine balance that had prevailed at the memory helper T cells priming to memorize the antigen As the priming event in this instance occurred amid inflammation the immune memory is pro-inflammatory
Acute and chronic transplant
rejection Therapy with protein A
Cellular immunity
As a cell is indicated by the prefix cyto a cytotoxicinfluence destroys the cell Alloreactive killer T cells also called cytotoxic T lymphocytes (CTLs) have CD8 receptors that dock to the transplanted tissues MHC class I moleculeswhich display the donors self peptides (In the living donor such presentation of self antigens helped maintain self tolerance) Thereupon the T cell receptors (TCRs) of the killer T cells recognize their matching epitope and trigger the target cells programmed cell death by apoptosis
Acute and chronic transplant
rejection Therapy with protein A
Humoral immunity
Developed through an earlier primary exposure that primed specific immunity to the nonself antigen a transplant recipient can have specific antibody crossreacting with the donor tissue upon the transplant event a secondary exposure This is typical after earlier mismatching among ABO blood types during blood transfusion At this secondary exposure these crossreactive antibody molecules interact with aspects of innate immunitymdashsoluble immune proteins called complement and innate immune cells calledphagocytesmdashwhich inflames and destroys the transplanted tissue
Acute and chronic transplant
rejection Therapy with protein A
Antibody[edit]
Secreted by an activated B cell then
called plasma cell an antibody molecule is a
soluble immunoglobulin (Ig) whose basic unit is
shaped like the letter Y the two arms are theFab
regions while the single stalk is the Fc region
Each of the two tips of Fab region is the paratope
which binds a matching molecular sequence and
its 3D shape (conformation) altogether
called epitope within the target antigen
Acute and chronic transplant
rejection Therapy with protein A
Opsonization
The IgGs Fc region also enables opsonization by a phagocyte a process by which the Fc receptor on the phagocytemdashsuch as neutrophils in blood and macrophages in tissuesmdashbinds the antibody molecules FC stalk and the phagocyte exhibits enhanced uptake of the antigen attached to the antibody molecules Fab region
Acute and chronic transplant
rejection Therapy with protein A
Complement cascade
When the paratope of Ig class gamma (IgG) binds its matching epitope IgGs Fc region conformationally shifts and can host a complement protein initiating the complement cascade that terminates by punching a hole in a cell membrane With many holes so punched fluid rushes into the cell and ruptures it
Cell debris can be recognized as damage associated molecular patterns (DAMPs) by pattern recognition receptors (PRRs) such as Toll-like receptors (TLRs) on membranes ofphagocytes which thereupon secrete proinflammatory cytokines recruiting more phagocytes to traffic to the area by sensing the concentration gradient of the secreted cytokines (chemotaxis)
Acute and chronic transplant
rejection Therapy with protein A
Protein A is a
42 kDa surface protein originally found in the
cell wall of the bacterium Staphylococcus
aureus It is encoded by the spa gene and its
regulation is controlled by DNA topology
cellular osmolarity and a two-component
system called ArlS-ArlR It has found use in
biochemical research because of its ability to
bind immunoglobulins
Acute and chronic transplant
rejection Therapy with protein A
It is composed of five homologous Ig-binding domains that fold into a three-helix bundle Each domain is able to bind proteins from many mammalian species most notably IgGs It binds the heavy chain within the Fc region of most immunoglobulins and also within the Fab region in the case of the human VH3 family Through these interactions in serum where IgG molecules are bound in the wrong orientation (in relation to normal antibodyfunction) the bacteria disrupts opsonization and phagocytosis
Acute and chronic transplant
rejection Therapy with protein A
Purified (unconjugated) Thermo Scientific Pierce Recombinant Protein A is useful as the basis for preparing various kinds of probes or affinity media for detection or purification of rabbit and human antibodies especially IgGisotypes in immunoassays and antibody purification protocols
Features of Recombinant Protein A
bull Contains four Fc-binding domains per proteinbull Better than Protein G for polyclonal IgG from rabbit pig dog and cat serumbull Poorer than Protein G for mouse IgG1 human IgG3 and rat goat and cow antibodies
Acute and chronic transplant
rejection Therapy with protein A
Protein A is a cell wall component produced by several strains of Staphylococcus aureus that consists of a single polypeptide chain and contains little or no carbohydrate Recombinant Protein A is produced in E coli and functions essentially the same as native Protein A The Protein A molecule contains four high-affinity binding sites capable of interacting with the Fcregion from IgG of several species including human and rabbit Optimal binding occurs at pH 82 although binding is also effective at neutral or physiological conditions (pH 70 to 76)
Acute and chronic transplant
rejection Therapy with protein A
httpswwwlifetechnologiescomordercatalog
product21184
Acute and chronic transplant
rejection Therapy with protein A
httpstoolslifetechnologiescomcontentsfsm
anualsMAN0011438_Pierce_Ig_Bind_Protein
s_UGpdf
httpstoolslifetechnologiescomcontentsfsbr
ochuresTR0034-Ab-binding-proteinspdf
Acute and chronic transplant
rejection Therapy with protein A
Crystal structure of a Staphylococcus
aureus protein A domain complexed with
the Fab fragment of a human IgM antibody
Structural basis for recognition of B-cell
receptors and superantigen activity Marc Graille et al Proc Natl Acad Sci U S A
2000 May 9 97(10) 5399ndash5404 PMCID PMC25840
Immunology
Acute and chronic transplant
rejection Therapy with protein A
Staphylococcus aureus Protein A Promotes
Immune Suppression
Scott D Kobayashi
Frank R DeLeo
Acute and chronic transplant
rejection Therapy with protein A
SpA binds the Fc region of antibody and the Fabregions of the B-cell receptor processes that are known to block opsonophagocytosis and cause B-cell death in vitro In a recent study Falugi et al [F Falugi H K Kim D M Missiakas and O Schneewind mBio4(5)e00575-13 2013] showed that vaccination with spa mutant S aureus strains lacking antibody Fc-andor Fab-binding capacity protects against subsequent challenge with the USA300 epidemic strain The findings provide strong support for the idea that SpA promotes S aureus immune evasion in vivo and form the foundation for a new approach in our efforts to develop a vaccine that prevents severe S aureusinfections
Acute and chronic transplant
rejection Therapy with protein A
Airflow obstruction not ascribable to other cause is labeled bronchiolitis obliterans syndrome (BOS) confirmed by a persistent dropmdashthree or more weeksmdashin forced expiratory volume (FEV1) by at least 20[17] BOS is seen in over 50 of lung-transplant recipients by 5 years and in over 80 by ten years First noted is infiltration by lymphocytes followed by epithelial cell injury then inflammatory lesions and recruitment of fibroblasts and myofibroblasts which proliferate and secrete proteins forming scar tissue
Acute and chronic transplant
rejection Therapy with protein A
Generally thought unpredictable BOS
progression varies widely lung function may
suddenly fall but stabilize for years or rapidly
progress to death within a few months Risk
factors include prior acute rejection
episodes gastroesophageal reflux disease acute
infections particular age groups HLA mis-
matching lymphocytic bronchiolitis and graft
dysfunction (eg airway ischemia)
httpenwikipediaorgwikiOrgan_transplantation
Acute and chronic transplant
rejection Therapy with protein A
Immune Mechanisms of Transplant Rejections
Rejection is an adaptive immune response via cellular immunity (mediated by killer T cells inducing apoptosis of target cells) as well as humoral immunity (mediated by activated B cells secreting antibody molecules) though the action is joined by components of innate immune response (phagocytes and soluble immune proteins) Different types of transplanted tissues tend to favor different balances of rejection mechanisms
Acute and chronic transplant
rejection Therapy with protein A
Immunization
An animals exposure to the antigens of a different member of the same or similar species is allostimulation and the tissue is allogenic Transplanted organs are often acquired from a cadaver (usually a host who had succumbed to trauma) whose tissues had already sustained ischemia or inflammation
Dendritic cells (DCs) which are the primary antigen-presenting cells (APCs) of the donor tissue migrate to the recipients peripheral lymphoid tissue (lymphoid follicles and lymph nodes) and present the donors self peptides to the recipients lymphocytes (immune cells residing in lymphoid tissues) Lymphocytes include two classes that enact adaptive immunity also called specific immunity Lymphocytes of specific immunity T cellsmdashincluding the subclasses helper T cells and killer T cellsmdashand B cells
The recipients helper T cells coordinate specific immunity directed at the donors self peptides or at the donors Major histocompatibilitycomplex molecules or at both
Acute and chronic transplant
rejection Therapy with protein A
Immune memory
When memory helper T cells CD4 receptors bind to the MHC class II molecules which are expressed on the surfaces of the target cells of the graft tissue the memory helper T cells T cell receptors (TCRs) can recognize their target antigen that is presented by the MHC class II molecules The memory helper T cell subsequently produces clones that as effector cells secrete immune signalling molecules (cytokines) in approximately the cytokine balance that had prevailed at the memory helper T cells priming to memorize the antigen As the priming event in this instance occurred amid inflammation the immune memory is pro-inflammatory
Acute and chronic transplant
rejection Therapy with protein A
Cellular immunity
As a cell is indicated by the prefix cyto a cytotoxicinfluence destroys the cell Alloreactive killer T cells also called cytotoxic T lymphocytes (CTLs) have CD8 receptors that dock to the transplanted tissues MHC class I moleculeswhich display the donors self peptides (In the living donor such presentation of self antigens helped maintain self tolerance) Thereupon the T cell receptors (TCRs) of the killer T cells recognize their matching epitope and trigger the target cells programmed cell death by apoptosis
Acute and chronic transplant
rejection Therapy with protein A
Humoral immunity
Developed through an earlier primary exposure that primed specific immunity to the nonself antigen a transplant recipient can have specific antibody crossreacting with the donor tissue upon the transplant event a secondary exposure This is typical after earlier mismatching among ABO blood types during blood transfusion At this secondary exposure these crossreactive antibody molecules interact with aspects of innate immunitymdashsoluble immune proteins called complement and innate immune cells calledphagocytesmdashwhich inflames and destroys the transplanted tissue
Acute and chronic transplant
rejection Therapy with protein A
Antibody[edit]
Secreted by an activated B cell then
called plasma cell an antibody molecule is a
soluble immunoglobulin (Ig) whose basic unit is
shaped like the letter Y the two arms are theFab
regions while the single stalk is the Fc region
Each of the two tips of Fab region is the paratope
which binds a matching molecular sequence and
its 3D shape (conformation) altogether
called epitope within the target antigen
Acute and chronic transplant
rejection Therapy with protein A
Opsonization
The IgGs Fc region also enables opsonization by a phagocyte a process by which the Fc receptor on the phagocytemdashsuch as neutrophils in blood and macrophages in tissuesmdashbinds the antibody molecules FC stalk and the phagocyte exhibits enhanced uptake of the antigen attached to the antibody molecules Fab region
Acute and chronic transplant
rejection Therapy with protein A
Complement cascade
When the paratope of Ig class gamma (IgG) binds its matching epitope IgGs Fc region conformationally shifts and can host a complement protein initiating the complement cascade that terminates by punching a hole in a cell membrane With many holes so punched fluid rushes into the cell and ruptures it
Cell debris can be recognized as damage associated molecular patterns (DAMPs) by pattern recognition receptors (PRRs) such as Toll-like receptors (TLRs) on membranes ofphagocytes which thereupon secrete proinflammatory cytokines recruiting more phagocytes to traffic to the area by sensing the concentration gradient of the secreted cytokines (chemotaxis)
Acute and chronic transplant
rejection Therapy with protein A
Protein A is a
42 kDa surface protein originally found in the
cell wall of the bacterium Staphylococcus
aureus It is encoded by the spa gene and its
regulation is controlled by DNA topology
cellular osmolarity and a two-component
system called ArlS-ArlR It has found use in
biochemical research because of its ability to
bind immunoglobulins
Acute and chronic transplant
rejection Therapy with protein A
It is composed of five homologous Ig-binding domains that fold into a three-helix bundle Each domain is able to bind proteins from many mammalian species most notably IgGs It binds the heavy chain within the Fc region of most immunoglobulins and also within the Fab region in the case of the human VH3 family Through these interactions in serum where IgG molecules are bound in the wrong orientation (in relation to normal antibodyfunction) the bacteria disrupts opsonization and phagocytosis
Acute and chronic transplant
rejection Therapy with protein A
Purified (unconjugated) Thermo Scientific Pierce Recombinant Protein A is useful as the basis for preparing various kinds of probes or affinity media for detection or purification of rabbit and human antibodies especially IgGisotypes in immunoassays and antibody purification protocols
Features of Recombinant Protein A
bull Contains four Fc-binding domains per proteinbull Better than Protein G for polyclonal IgG from rabbit pig dog and cat serumbull Poorer than Protein G for mouse IgG1 human IgG3 and rat goat and cow antibodies
Acute and chronic transplant
rejection Therapy with protein A
Protein A is a cell wall component produced by several strains of Staphylococcus aureus that consists of a single polypeptide chain and contains little or no carbohydrate Recombinant Protein A is produced in E coli and functions essentially the same as native Protein A The Protein A molecule contains four high-affinity binding sites capable of interacting with the Fcregion from IgG of several species including human and rabbit Optimal binding occurs at pH 82 although binding is also effective at neutral or physiological conditions (pH 70 to 76)
Acute and chronic transplant
rejection Therapy with protein A
httpswwwlifetechnologiescomordercatalog
product21184
Acute and chronic transplant
rejection Therapy with protein A
httpstoolslifetechnologiescomcontentsfsm
anualsMAN0011438_Pierce_Ig_Bind_Protein
s_UGpdf
httpstoolslifetechnologiescomcontentsfsbr
ochuresTR0034-Ab-binding-proteinspdf
Acute and chronic transplant
rejection Therapy with protein A
Crystal structure of a Staphylococcus
aureus protein A domain complexed with
the Fab fragment of a human IgM antibody
Structural basis for recognition of B-cell
receptors and superantigen activity Marc Graille et al Proc Natl Acad Sci U S A
2000 May 9 97(10) 5399ndash5404 PMCID PMC25840
Immunology
Acute and chronic transplant
rejection Therapy with protein A
Staphylococcus aureus Protein A Promotes
Immune Suppression
Scott D Kobayashi
Frank R DeLeo
Acute and chronic transplant
rejection Therapy with protein A
SpA binds the Fc region of antibody and the Fabregions of the B-cell receptor processes that are known to block opsonophagocytosis and cause B-cell death in vitro In a recent study Falugi et al [F Falugi H K Kim D M Missiakas and O Schneewind mBio4(5)e00575-13 2013] showed that vaccination with spa mutant S aureus strains lacking antibody Fc-andor Fab-binding capacity protects against subsequent challenge with the USA300 epidemic strain The findings provide strong support for the idea that SpA promotes S aureus immune evasion in vivo and form the foundation for a new approach in our efforts to develop a vaccine that prevents severe S aureusinfections
Acute and chronic transplant
rejection Therapy with protein A
Generally thought unpredictable BOS
progression varies widely lung function may
suddenly fall but stabilize for years or rapidly
progress to death within a few months Risk
factors include prior acute rejection
episodes gastroesophageal reflux disease acute
infections particular age groups HLA mis-
matching lymphocytic bronchiolitis and graft
dysfunction (eg airway ischemia)
httpenwikipediaorgwikiOrgan_transplantation
Acute and chronic transplant
rejection Therapy with protein A
Immune Mechanisms of Transplant Rejections
Rejection is an adaptive immune response via cellular immunity (mediated by killer T cells inducing apoptosis of target cells) as well as humoral immunity (mediated by activated B cells secreting antibody molecules) though the action is joined by components of innate immune response (phagocytes and soluble immune proteins) Different types of transplanted tissues tend to favor different balances of rejection mechanisms
Acute and chronic transplant
rejection Therapy with protein A
Immunization
An animals exposure to the antigens of a different member of the same or similar species is allostimulation and the tissue is allogenic Transplanted organs are often acquired from a cadaver (usually a host who had succumbed to trauma) whose tissues had already sustained ischemia or inflammation
Dendritic cells (DCs) which are the primary antigen-presenting cells (APCs) of the donor tissue migrate to the recipients peripheral lymphoid tissue (lymphoid follicles and lymph nodes) and present the donors self peptides to the recipients lymphocytes (immune cells residing in lymphoid tissues) Lymphocytes include two classes that enact adaptive immunity also called specific immunity Lymphocytes of specific immunity T cellsmdashincluding the subclasses helper T cells and killer T cellsmdashand B cells
The recipients helper T cells coordinate specific immunity directed at the donors self peptides or at the donors Major histocompatibilitycomplex molecules or at both
Acute and chronic transplant
rejection Therapy with protein A
Immune memory
When memory helper T cells CD4 receptors bind to the MHC class II molecules which are expressed on the surfaces of the target cells of the graft tissue the memory helper T cells T cell receptors (TCRs) can recognize their target antigen that is presented by the MHC class II molecules The memory helper T cell subsequently produces clones that as effector cells secrete immune signalling molecules (cytokines) in approximately the cytokine balance that had prevailed at the memory helper T cells priming to memorize the antigen As the priming event in this instance occurred amid inflammation the immune memory is pro-inflammatory
Acute and chronic transplant
rejection Therapy with protein A
Cellular immunity
As a cell is indicated by the prefix cyto a cytotoxicinfluence destroys the cell Alloreactive killer T cells also called cytotoxic T lymphocytes (CTLs) have CD8 receptors that dock to the transplanted tissues MHC class I moleculeswhich display the donors self peptides (In the living donor such presentation of self antigens helped maintain self tolerance) Thereupon the T cell receptors (TCRs) of the killer T cells recognize their matching epitope and trigger the target cells programmed cell death by apoptosis
Acute and chronic transplant
rejection Therapy with protein A
Humoral immunity
Developed through an earlier primary exposure that primed specific immunity to the nonself antigen a transplant recipient can have specific antibody crossreacting with the donor tissue upon the transplant event a secondary exposure This is typical after earlier mismatching among ABO blood types during blood transfusion At this secondary exposure these crossreactive antibody molecules interact with aspects of innate immunitymdashsoluble immune proteins called complement and innate immune cells calledphagocytesmdashwhich inflames and destroys the transplanted tissue
Acute and chronic transplant
rejection Therapy with protein A
Antibody[edit]
Secreted by an activated B cell then
called plasma cell an antibody molecule is a
soluble immunoglobulin (Ig) whose basic unit is
shaped like the letter Y the two arms are theFab
regions while the single stalk is the Fc region
Each of the two tips of Fab region is the paratope
which binds a matching molecular sequence and
its 3D shape (conformation) altogether
called epitope within the target antigen
Acute and chronic transplant
rejection Therapy with protein A
Opsonization
The IgGs Fc region also enables opsonization by a phagocyte a process by which the Fc receptor on the phagocytemdashsuch as neutrophils in blood and macrophages in tissuesmdashbinds the antibody molecules FC stalk and the phagocyte exhibits enhanced uptake of the antigen attached to the antibody molecules Fab region
Acute and chronic transplant
rejection Therapy with protein A
Complement cascade
When the paratope of Ig class gamma (IgG) binds its matching epitope IgGs Fc region conformationally shifts and can host a complement protein initiating the complement cascade that terminates by punching a hole in a cell membrane With many holes so punched fluid rushes into the cell and ruptures it
Cell debris can be recognized as damage associated molecular patterns (DAMPs) by pattern recognition receptors (PRRs) such as Toll-like receptors (TLRs) on membranes ofphagocytes which thereupon secrete proinflammatory cytokines recruiting more phagocytes to traffic to the area by sensing the concentration gradient of the secreted cytokines (chemotaxis)
Acute and chronic transplant
rejection Therapy with protein A
Protein A is a
42 kDa surface protein originally found in the
cell wall of the bacterium Staphylococcus
aureus It is encoded by the spa gene and its
regulation is controlled by DNA topology
cellular osmolarity and a two-component
system called ArlS-ArlR It has found use in
biochemical research because of its ability to
bind immunoglobulins
Acute and chronic transplant
rejection Therapy with protein A
It is composed of five homologous Ig-binding domains that fold into a three-helix bundle Each domain is able to bind proteins from many mammalian species most notably IgGs It binds the heavy chain within the Fc region of most immunoglobulins and also within the Fab region in the case of the human VH3 family Through these interactions in serum where IgG molecules are bound in the wrong orientation (in relation to normal antibodyfunction) the bacteria disrupts opsonization and phagocytosis
Acute and chronic transplant
rejection Therapy with protein A
Purified (unconjugated) Thermo Scientific Pierce Recombinant Protein A is useful as the basis for preparing various kinds of probes or affinity media for detection or purification of rabbit and human antibodies especially IgGisotypes in immunoassays and antibody purification protocols
Features of Recombinant Protein A
bull Contains four Fc-binding domains per proteinbull Better than Protein G for polyclonal IgG from rabbit pig dog and cat serumbull Poorer than Protein G for mouse IgG1 human IgG3 and rat goat and cow antibodies
Acute and chronic transplant
rejection Therapy with protein A
Protein A is a cell wall component produced by several strains of Staphylococcus aureus that consists of a single polypeptide chain and contains little or no carbohydrate Recombinant Protein A is produced in E coli and functions essentially the same as native Protein A The Protein A molecule contains four high-affinity binding sites capable of interacting with the Fcregion from IgG of several species including human and rabbit Optimal binding occurs at pH 82 although binding is also effective at neutral or physiological conditions (pH 70 to 76)
Acute and chronic transplant
rejection Therapy with protein A
httpswwwlifetechnologiescomordercatalog
product21184
Acute and chronic transplant
rejection Therapy with protein A
httpstoolslifetechnologiescomcontentsfsm
anualsMAN0011438_Pierce_Ig_Bind_Protein
s_UGpdf
httpstoolslifetechnologiescomcontentsfsbr
ochuresTR0034-Ab-binding-proteinspdf
Acute and chronic transplant
rejection Therapy with protein A
Crystal structure of a Staphylococcus
aureus protein A domain complexed with
the Fab fragment of a human IgM antibody
Structural basis for recognition of B-cell
receptors and superantigen activity Marc Graille et al Proc Natl Acad Sci U S A
2000 May 9 97(10) 5399ndash5404 PMCID PMC25840
Immunology
Acute and chronic transplant
rejection Therapy with protein A
Staphylococcus aureus Protein A Promotes
Immune Suppression
Scott D Kobayashi
Frank R DeLeo
Acute and chronic transplant
rejection Therapy with protein A
SpA binds the Fc region of antibody and the Fabregions of the B-cell receptor processes that are known to block opsonophagocytosis and cause B-cell death in vitro In a recent study Falugi et al [F Falugi H K Kim D M Missiakas and O Schneewind mBio4(5)e00575-13 2013] showed that vaccination with spa mutant S aureus strains lacking antibody Fc-andor Fab-binding capacity protects against subsequent challenge with the USA300 epidemic strain The findings provide strong support for the idea that SpA promotes S aureus immune evasion in vivo and form the foundation for a new approach in our efforts to develop a vaccine that prevents severe S aureusinfections
Acute and chronic transplant
rejection Therapy with protein A
Immune Mechanisms of Transplant Rejections
Rejection is an adaptive immune response via cellular immunity (mediated by killer T cells inducing apoptosis of target cells) as well as humoral immunity (mediated by activated B cells secreting antibody molecules) though the action is joined by components of innate immune response (phagocytes and soluble immune proteins) Different types of transplanted tissues tend to favor different balances of rejection mechanisms
Acute and chronic transplant
rejection Therapy with protein A
Immunization
An animals exposure to the antigens of a different member of the same or similar species is allostimulation and the tissue is allogenic Transplanted organs are often acquired from a cadaver (usually a host who had succumbed to trauma) whose tissues had already sustained ischemia or inflammation
Dendritic cells (DCs) which are the primary antigen-presenting cells (APCs) of the donor tissue migrate to the recipients peripheral lymphoid tissue (lymphoid follicles and lymph nodes) and present the donors self peptides to the recipients lymphocytes (immune cells residing in lymphoid tissues) Lymphocytes include two classes that enact adaptive immunity also called specific immunity Lymphocytes of specific immunity T cellsmdashincluding the subclasses helper T cells and killer T cellsmdashand B cells
The recipients helper T cells coordinate specific immunity directed at the donors self peptides or at the donors Major histocompatibilitycomplex molecules or at both
Acute and chronic transplant
rejection Therapy with protein A
Immune memory
When memory helper T cells CD4 receptors bind to the MHC class II molecules which are expressed on the surfaces of the target cells of the graft tissue the memory helper T cells T cell receptors (TCRs) can recognize their target antigen that is presented by the MHC class II molecules The memory helper T cell subsequently produces clones that as effector cells secrete immune signalling molecules (cytokines) in approximately the cytokine balance that had prevailed at the memory helper T cells priming to memorize the antigen As the priming event in this instance occurred amid inflammation the immune memory is pro-inflammatory
Acute and chronic transplant
rejection Therapy with protein A
Cellular immunity
As a cell is indicated by the prefix cyto a cytotoxicinfluence destroys the cell Alloreactive killer T cells also called cytotoxic T lymphocytes (CTLs) have CD8 receptors that dock to the transplanted tissues MHC class I moleculeswhich display the donors self peptides (In the living donor such presentation of self antigens helped maintain self tolerance) Thereupon the T cell receptors (TCRs) of the killer T cells recognize their matching epitope and trigger the target cells programmed cell death by apoptosis
Acute and chronic transplant
rejection Therapy with protein A
Humoral immunity
Developed through an earlier primary exposure that primed specific immunity to the nonself antigen a transplant recipient can have specific antibody crossreacting with the donor tissue upon the transplant event a secondary exposure This is typical after earlier mismatching among ABO blood types during blood transfusion At this secondary exposure these crossreactive antibody molecules interact with aspects of innate immunitymdashsoluble immune proteins called complement and innate immune cells calledphagocytesmdashwhich inflames and destroys the transplanted tissue
Acute and chronic transplant
rejection Therapy with protein A
Antibody[edit]
Secreted by an activated B cell then
called plasma cell an antibody molecule is a
soluble immunoglobulin (Ig) whose basic unit is
shaped like the letter Y the two arms are theFab
regions while the single stalk is the Fc region
Each of the two tips of Fab region is the paratope
which binds a matching molecular sequence and
its 3D shape (conformation) altogether
called epitope within the target antigen
Acute and chronic transplant
rejection Therapy with protein A
Opsonization
The IgGs Fc region also enables opsonization by a phagocyte a process by which the Fc receptor on the phagocytemdashsuch as neutrophils in blood and macrophages in tissuesmdashbinds the antibody molecules FC stalk and the phagocyte exhibits enhanced uptake of the antigen attached to the antibody molecules Fab region
Acute and chronic transplant
rejection Therapy with protein A
Complement cascade
When the paratope of Ig class gamma (IgG) binds its matching epitope IgGs Fc region conformationally shifts and can host a complement protein initiating the complement cascade that terminates by punching a hole in a cell membrane With many holes so punched fluid rushes into the cell and ruptures it
Cell debris can be recognized as damage associated molecular patterns (DAMPs) by pattern recognition receptors (PRRs) such as Toll-like receptors (TLRs) on membranes ofphagocytes which thereupon secrete proinflammatory cytokines recruiting more phagocytes to traffic to the area by sensing the concentration gradient of the secreted cytokines (chemotaxis)
Acute and chronic transplant
rejection Therapy with protein A
Protein A is a
42 kDa surface protein originally found in the
cell wall of the bacterium Staphylococcus
aureus It is encoded by the spa gene and its
regulation is controlled by DNA topology
cellular osmolarity and a two-component
system called ArlS-ArlR It has found use in
biochemical research because of its ability to
bind immunoglobulins
Acute and chronic transplant
rejection Therapy with protein A
It is composed of five homologous Ig-binding domains that fold into a three-helix bundle Each domain is able to bind proteins from many mammalian species most notably IgGs It binds the heavy chain within the Fc region of most immunoglobulins and also within the Fab region in the case of the human VH3 family Through these interactions in serum where IgG molecules are bound in the wrong orientation (in relation to normal antibodyfunction) the bacteria disrupts opsonization and phagocytosis
Acute and chronic transplant
rejection Therapy with protein A
Purified (unconjugated) Thermo Scientific Pierce Recombinant Protein A is useful as the basis for preparing various kinds of probes or affinity media for detection or purification of rabbit and human antibodies especially IgGisotypes in immunoassays and antibody purification protocols
Features of Recombinant Protein A
bull Contains four Fc-binding domains per proteinbull Better than Protein G for polyclonal IgG from rabbit pig dog and cat serumbull Poorer than Protein G for mouse IgG1 human IgG3 and rat goat and cow antibodies
Acute and chronic transplant
rejection Therapy with protein A
Protein A is a cell wall component produced by several strains of Staphylococcus aureus that consists of a single polypeptide chain and contains little or no carbohydrate Recombinant Protein A is produced in E coli and functions essentially the same as native Protein A The Protein A molecule contains four high-affinity binding sites capable of interacting with the Fcregion from IgG of several species including human and rabbit Optimal binding occurs at pH 82 although binding is also effective at neutral or physiological conditions (pH 70 to 76)
Acute and chronic transplant
rejection Therapy with protein A
httpswwwlifetechnologiescomordercatalog
product21184
Acute and chronic transplant
rejection Therapy with protein A
httpstoolslifetechnologiescomcontentsfsm
anualsMAN0011438_Pierce_Ig_Bind_Protein
s_UGpdf
httpstoolslifetechnologiescomcontentsfsbr
ochuresTR0034-Ab-binding-proteinspdf
Acute and chronic transplant
rejection Therapy with protein A
Crystal structure of a Staphylococcus
aureus protein A domain complexed with
the Fab fragment of a human IgM antibody
Structural basis for recognition of B-cell
receptors and superantigen activity Marc Graille et al Proc Natl Acad Sci U S A
2000 May 9 97(10) 5399ndash5404 PMCID PMC25840
Immunology
Acute and chronic transplant
rejection Therapy with protein A
Staphylococcus aureus Protein A Promotes
Immune Suppression
Scott D Kobayashi
Frank R DeLeo
Acute and chronic transplant
rejection Therapy with protein A
SpA binds the Fc region of antibody and the Fabregions of the B-cell receptor processes that are known to block opsonophagocytosis and cause B-cell death in vitro In a recent study Falugi et al [F Falugi H K Kim D M Missiakas and O Schneewind mBio4(5)e00575-13 2013] showed that vaccination with spa mutant S aureus strains lacking antibody Fc-andor Fab-binding capacity protects against subsequent challenge with the USA300 epidemic strain The findings provide strong support for the idea that SpA promotes S aureus immune evasion in vivo and form the foundation for a new approach in our efforts to develop a vaccine that prevents severe S aureusinfections
Acute and chronic transplant
rejection Therapy with protein A
Immunization
An animals exposure to the antigens of a different member of the same or similar species is allostimulation and the tissue is allogenic Transplanted organs are often acquired from a cadaver (usually a host who had succumbed to trauma) whose tissues had already sustained ischemia or inflammation
Dendritic cells (DCs) which are the primary antigen-presenting cells (APCs) of the donor tissue migrate to the recipients peripheral lymphoid tissue (lymphoid follicles and lymph nodes) and present the donors self peptides to the recipients lymphocytes (immune cells residing in lymphoid tissues) Lymphocytes include two classes that enact adaptive immunity also called specific immunity Lymphocytes of specific immunity T cellsmdashincluding the subclasses helper T cells and killer T cellsmdashand B cells
The recipients helper T cells coordinate specific immunity directed at the donors self peptides or at the donors Major histocompatibilitycomplex molecules or at both
Acute and chronic transplant
rejection Therapy with protein A
Immune memory
When memory helper T cells CD4 receptors bind to the MHC class II molecules which are expressed on the surfaces of the target cells of the graft tissue the memory helper T cells T cell receptors (TCRs) can recognize their target antigen that is presented by the MHC class II molecules The memory helper T cell subsequently produces clones that as effector cells secrete immune signalling molecules (cytokines) in approximately the cytokine balance that had prevailed at the memory helper T cells priming to memorize the antigen As the priming event in this instance occurred amid inflammation the immune memory is pro-inflammatory
Acute and chronic transplant
rejection Therapy with protein A
Cellular immunity
As a cell is indicated by the prefix cyto a cytotoxicinfluence destroys the cell Alloreactive killer T cells also called cytotoxic T lymphocytes (CTLs) have CD8 receptors that dock to the transplanted tissues MHC class I moleculeswhich display the donors self peptides (In the living donor such presentation of self antigens helped maintain self tolerance) Thereupon the T cell receptors (TCRs) of the killer T cells recognize their matching epitope and trigger the target cells programmed cell death by apoptosis
Acute and chronic transplant
rejection Therapy with protein A
Humoral immunity
Developed through an earlier primary exposure that primed specific immunity to the nonself antigen a transplant recipient can have specific antibody crossreacting with the donor tissue upon the transplant event a secondary exposure This is typical after earlier mismatching among ABO blood types during blood transfusion At this secondary exposure these crossreactive antibody molecules interact with aspects of innate immunitymdashsoluble immune proteins called complement and innate immune cells calledphagocytesmdashwhich inflames and destroys the transplanted tissue
Acute and chronic transplant
rejection Therapy with protein A
Antibody[edit]
Secreted by an activated B cell then
called plasma cell an antibody molecule is a
soluble immunoglobulin (Ig) whose basic unit is
shaped like the letter Y the two arms are theFab
regions while the single stalk is the Fc region
Each of the two tips of Fab region is the paratope
which binds a matching molecular sequence and
its 3D shape (conformation) altogether
called epitope within the target antigen
Acute and chronic transplant
rejection Therapy with protein A
Opsonization
The IgGs Fc region also enables opsonization by a phagocyte a process by which the Fc receptor on the phagocytemdashsuch as neutrophils in blood and macrophages in tissuesmdashbinds the antibody molecules FC stalk and the phagocyte exhibits enhanced uptake of the antigen attached to the antibody molecules Fab region
Acute and chronic transplant
rejection Therapy with protein A
Complement cascade
When the paratope of Ig class gamma (IgG) binds its matching epitope IgGs Fc region conformationally shifts and can host a complement protein initiating the complement cascade that terminates by punching a hole in a cell membrane With many holes so punched fluid rushes into the cell and ruptures it
Cell debris can be recognized as damage associated molecular patterns (DAMPs) by pattern recognition receptors (PRRs) such as Toll-like receptors (TLRs) on membranes ofphagocytes which thereupon secrete proinflammatory cytokines recruiting more phagocytes to traffic to the area by sensing the concentration gradient of the secreted cytokines (chemotaxis)
Acute and chronic transplant
rejection Therapy with protein A
Protein A is a
42 kDa surface protein originally found in the
cell wall of the bacterium Staphylococcus
aureus It is encoded by the spa gene and its
regulation is controlled by DNA topology
cellular osmolarity and a two-component
system called ArlS-ArlR It has found use in
biochemical research because of its ability to
bind immunoglobulins
Acute and chronic transplant
rejection Therapy with protein A
It is composed of five homologous Ig-binding domains that fold into a three-helix bundle Each domain is able to bind proteins from many mammalian species most notably IgGs It binds the heavy chain within the Fc region of most immunoglobulins and also within the Fab region in the case of the human VH3 family Through these interactions in serum where IgG molecules are bound in the wrong orientation (in relation to normal antibodyfunction) the bacteria disrupts opsonization and phagocytosis
Acute and chronic transplant
rejection Therapy with protein A
Purified (unconjugated) Thermo Scientific Pierce Recombinant Protein A is useful as the basis for preparing various kinds of probes or affinity media for detection or purification of rabbit and human antibodies especially IgGisotypes in immunoassays and antibody purification protocols
Features of Recombinant Protein A
bull Contains four Fc-binding domains per proteinbull Better than Protein G for polyclonal IgG from rabbit pig dog and cat serumbull Poorer than Protein G for mouse IgG1 human IgG3 and rat goat and cow antibodies
Acute and chronic transplant
rejection Therapy with protein A
Protein A is a cell wall component produced by several strains of Staphylococcus aureus that consists of a single polypeptide chain and contains little or no carbohydrate Recombinant Protein A is produced in E coli and functions essentially the same as native Protein A The Protein A molecule contains four high-affinity binding sites capable of interacting with the Fcregion from IgG of several species including human and rabbit Optimal binding occurs at pH 82 although binding is also effective at neutral or physiological conditions (pH 70 to 76)
Acute and chronic transplant
rejection Therapy with protein A
httpswwwlifetechnologiescomordercatalog
product21184
Acute and chronic transplant
rejection Therapy with protein A
httpstoolslifetechnologiescomcontentsfsm
anualsMAN0011438_Pierce_Ig_Bind_Protein
s_UGpdf
httpstoolslifetechnologiescomcontentsfsbr
ochuresTR0034-Ab-binding-proteinspdf
Acute and chronic transplant
rejection Therapy with protein A
Crystal structure of a Staphylococcus
aureus protein A domain complexed with
the Fab fragment of a human IgM antibody
Structural basis for recognition of B-cell
receptors and superantigen activity Marc Graille et al Proc Natl Acad Sci U S A
2000 May 9 97(10) 5399ndash5404 PMCID PMC25840
Immunology
Acute and chronic transplant
rejection Therapy with protein A
Staphylococcus aureus Protein A Promotes
Immune Suppression
Scott D Kobayashi
Frank R DeLeo
Acute and chronic transplant
rejection Therapy with protein A
SpA binds the Fc region of antibody and the Fabregions of the B-cell receptor processes that are known to block opsonophagocytosis and cause B-cell death in vitro In a recent study Falugi et al [F Falugi H K Kim D M Missiakas and O Schneewind mBio4(5)e00575-13 2013] showed that vaccination with spa mutant S aureus strains lacking antibody Fc-andor Fab-binding capacity protects against subsequent challenge with the USA300 epidemic strain The findings provide strong support for the idea that SpA promotes S aureus immune evasion in vivo and form the foundation for a new approach in our efforts to develop a vaccine that prevents severe S aureusinfections
Acute and chronic transplant
rejection Therapy with protein A
Immune memory
When memory helper T cells CD4 receptors bind to the MHC class II molecules which are expressed on the surfaces of the target cells of the graft tissue the memory helper T cells T cell receptors (TCRs) can recognize their target antigen that is presented by the MHC class II molecules The memory helper T cell subsequently produces clones that as effector cells secrete immune signalling molecules (cytokines) in approximately the cytokine balance that had prevailed at the memory helper T cells priming to memorize the antigen As the priming event in this instance occurred amid inflammation the immune memory is pro-inflammatory
Acute and chronic transplant
rejection Therapy with protein A
Cellular immunity
As a cell is indicated by the prefix cyto a cytotoxicinfluence destroys the cell Alloreactive killer T cells also called cytotoxic T lymphocytes (CTLs) have CD8 receptors that dock to the transplanted tissues MHC class I moleculeswhich display the donors self peptides (In the living donor such presentation of self antigens helped maintain self tolerance) Thereupon the T cell receptors (TCRs) of the killer T cells recognize their matching epitope and trigger the target cells programmed cell death by apoptosis
Acute and chronic transplant
rejection Therapy with protein A
Humoral immunity
Developed through an earlier primary exposure that primed specific immunity to the nonself antigen a transplant recipient can have specific antibody crossreacting with the donor tissue upon the transplant event a secondary exposure This is typical after earlier mismatching among ABO blood types during blood transfusion At this secondary exposure these crossreactive antibody molecules interact with aspects of innate immunitymdashsoluble immune proteins called complement and innate immune cells calledphagocytesmdashwhich inflames and destroys the transplanted tissue
Acute and chronic transplant
rejection Therapy with protein A
Antibody[edit]
Secreted by an activated B cell then
called plasma cell an antibody molecule is a
soluble immunoglobulin (Ig) whose basic unit is
shaped like the letter Y the two arms are theFab
regions while the single stalk is the Fc region
Each of the two tips of Fab region is the paratope
which binds a matching molecular sequence and
its 3D shape (conformation) altogether
called epitope within the target antigen
Acute and chronic transplant
rejection Therapy with protein A
Opsonization
The IgGs Fc region also enables opsonization by a phagocyte a process by which the Fc receptor on the phagocytemdashsuch as neutrophils in blood and macrophages in tissuesmdashbinds the antibody molecules FC stalk and the phagocyte exhibits enhanced uptake of the antigen attached to the antibody molecules Fab region
Acute and chronic transplant
rejection Therapy with protein A
Complement cascade
When the paratope of Ig class gamma (IgG) binds its matching epitope IgGs Fc region conformationally shifts and can host a complement protein initiating the complement cascade that terminates by punching a hole in a cell membrane With many holes so punched fluid rushes into the cell and ruptures it
Cell debris can be recognized as damage associated molecular patterns (DAMPs) by pattern recognition receptors (PRRs) such as Toll-like receptors (TLRs) on membranes ofphagocytes which thereupon secrete proinflammatory cytokines recruiting more phagocytes to traffic to the area by sensing the concentration gradient of the secreted cytokines (chemotaxis)
Acute and chronic transplant
rejection Therapy with protein A
Protein A is a
42 kDa surface protein originally found in the
cell wall of the bacterium Staphylococcus
aureus It is encoded by the spa gene and its
regulation is controlled by DNA topology
cellular osmolarity and a two-component
system called ArlS-ArlR It has found use in
biochemical research because of its ability to
bind immunoglobulins
Acute and chronic transplant
rejection Therapy with protein A
It is composed of five homologous Ig-binding domains that fold into a three-helix bundle Each domain is able to bind proteins from many mammalian species most notably IgGs It binds the heavy chain within the Fc region of most immunoglobulins and also within the Fab region in the case of the human VH3 family Through these interactions in serum where IgG molecules are bound in the wrong orientation (in relation to normal antibodyfunction) the bacteria disrupts opsonization and phagocytosis
Acute and chronic transplant
rejection Therapy with protein A
Purified (unconjugated) Thermo Scientific Pierce Recombinant Protein A is useful as the basis for preparing various kinds of probes or affinity media for detection or purification of rabbit and human antibodies especially IgGisotypes in immunoassays and antibody purification protocols
Features of Recombinant Protein A
bull Contains four Fc-binding domains per proteinbull Better than Protein G for polyclonal IgG from rabbit pig dog and cat serumbull Poorer than Protein G for mouse IgG1 human IgG3 and rat goat and cow antibodies
Acute and chronic transplant
rejection Therapy with protein A
Protein A is a cell wall component produced by several strains of Staphylococcus aureus that consists of a single polypeptide chain and contains little or no carbohydrate Recombinant Protein A is produced in E coli and functions essentially the same as native Protein A The Protein A molecule contains four high-affinity binding sites capable of interacting with the Fcregion from IgG of several species including human and rabbit Optimal binding occurs at pH 82 although binding is also effective at neutral or physiological conditions (pH 70 to 76)
Acute and chronic transplant
rejection Therapy with protein A
httpswwwlifetechnologiescomordercatalog
product21184
Acute and chronic transplant
rejection Therapy with protein A
httpstoolslifetechnologiescomcontentsfsm
anualsMAN0011438_Pierce_Ig_Bind_Protein
s_UGpdf
httpstoolslifetechnologiescomcontentsfsbr
ochuresTR0034-Ab-binding-proteinspdf
Acute and chronic transplant
rejection Therapy with protein A
Crystal structure of a Staphylococcus
aureus protein A domain complexed with
the Fab fragment of a human IgM antibody
Structural basis for recognition of B-cell
receptors and superantigen activity Marc Graille et al Proc Natl Acad Sci U S A
2000 May 9 97(10) 5399ndash5404 PMCID PMC25840
Immunology
Acute and chronic transplant
rejection Therapy with protein A
Staphylococcus aureus Protein A Promotes
Immune Suppression
Scott D Kobayashi
Frank R DeLeo
Acute and chronic transplant
rejection Therapy with protein A
SpA binds the Fc region of antibody and the Fabregions of the B-cell receptor processes that are known to block opsonophagocytosis and cause B-cell death in vitro In a recent study Falugi et al [F Falugi H K Kim D M Missiakas and O Schneewind mBio4(5)e00575-13 2013] showed that vaccination with spa mutant S aureus strains lacking antibody Fc-andor Fab-binding capacity protects against subsequent challenge with the USA300 epidemic strain The findings provide strong support for the idea that SpA promotes S aureus immune evasion in vivo and form the foundation for a new approach in our efforts to develop a vaccine that prevents severe S aureusinfections
Acute and chronic transplant
rejection Therapy with protein A
Cellular immunity
As a cell is indicated by the prefix cyto a cytotoxicinfluence destroys the cell Alloreactive killer T cells also called cytotoxic T lymphocytes (CTLs) have CD8 receptors that dock to the transplanted tissues MHC class I moleculeswhich display the donors self peptides (In the living donor such presentation of self antigens helped maintain self tolerance) Thereupon the T cell receptors (TCRs) of the killer T cells recognize their matching epitope and trigger the target cells programmed cell death by apoptosis
Acute and chronic transplant
rejection Therapy with protein A
Humoral immunity
Developed through an earlier primary exposure that primed specific immunity to the nonself antigen a transplant recipient can have specific antibody crossreacting with the donor tissue upon the transplant event a secondary exposure This is typical after earlier mismatching among ABO blood types during blood transfusion At this secondary exposure these crossreactive antibody molecules interact with aspects of innate immunitymdashsoluble immune proteins called complement and innate immune cells calledphagocytesmdashwhich inflames and destroys the transplanted tissue
Acute and chronic transplant
rejection Therapy with protein A
Antibody[edit]
Secreted by an activated B cell then
called plasma cell an antibody molecule is a
soluble immunoglobulin (Ig) whose basic unit is
shaped like the letter Y the two arms are theFab
regions while the single stalk is the Fc region
Each of the two tips of Fab region is the paratope
which binds a matching molecular sequence and
its 3D shape (conformation) altogether
called epitope within the target antigen
Acute and chronic transplant
rejection Therapy with protein A
Opsonization
The IgGs Fc region also enables opsonization by a phagocyte a process by which the Fc receptor on the phagocytemdashsuch as neutrophils in blood and macrophages in tissuesmdashbinds the antibody molecules FC stalk and the phagocyte exhibits enhanced uptake of the antigen attached to the antibody molecules Fab region
Acute and chronic transplant
rejection Therapy with protein A
Complement cascade
When the paratope of Ig class gamma (IgG) binds its matching epitope IgGs Fc region conformationally shifts and can host a complement protein initiating the complement cascade that terminates by punching a hole in a cell membrane With many holes so punched fluid rushes into the cell and ruptures it
Cell debris can be recognized as damage associated molecular patterns (DAMPs) by pattern recognition receptors (PRRs) such as Toll-like receptors (TLRs) on membranes ofphagocytes which thereupon secrete proinflammatory cytokines recruiting more phagocytes to traffic to the area by sensing the concentration gradient of the secreted cytokines (chemotaxis)
Acute and chronic transplant
rejection Therapy with protein A
Protein A is a
42 kDa surface protein originally found in the
cell wall of the bacterium Staphylococcus
aureus It is encoded by the spa gene and its
regulation is controlled by DNA topology
cellular osmolarity and a two-component
system called ArlS-ArlR It has found use in
biochemical research because of its ability to
bind immunoglobulins
Acute and chronic transplant
rejection Therapy with protein A
It is composed of five homologous Ig-binding domains that fold into a three-helix bundle Each domain is able to bind proteins from many mammalian species most notably IgGs It binds the heavy chain within the Fc region of most immunoglobulins and also within the Fab region in the case of the human VH3 family Through these interactions in serum where IgG molecules are bound in the wrong orientation (in relation to normal antibodyfunction) the bacteria disrupts opsonization and phagocytosis
Acute and chronic transplant
rejection Therapy with protein A
Purified (unconjugated) Thermo Scientific Pierce Recombinant Protein A is useful as the basis for preparing various kinds of probes or affinity media for detection or purification of rabbit and human antibodies especially IgGisotypes in immunoassays and antibody purification protocols
Features of Recombinant Protein A
bull Contains four Fc-binding domains per proteinbull Better than Protein G for polyclonal IgG from rabbit pig dog and cat serumbull Poorer than Protein G for mouse IgG1 human IgG3 and rat goat and cow antibodies
Acute and chronic transplant
rejection Therapy with protein A
Protein A is a cell wall component produced by several strains of Staphylococcus aureus that consists of a single polypeptide chain and contains little or no carbohydrate Recombinant Protein A is produced in E coli and functions essentially the same as native Protein A The Protein A molecule contains four high-affinity binding sites capable of interacting with the Fcregion from IgG of several species including human and rabbit Optimal binding occurs at pH 82 although binding is also effective at neutral or physiological conditions (pH 70 to 76)
Acute and chronic transplant
rejection Therapy with protein A
httpswwwlifetechnologiescomordercatalog
product21184
Acute and chronic transplant
rejection Therapy with protein A
httpstoolslifetechnologiescomcontentsfsm
anualsMAN0011438_Pierce_Ig_Bind_Protein
s_UGpdf
httpstoolslifetechnologiescomcontentsfsbr
ochuresTR0034-Ab-binding-proteinspdf
Acute and chronic transplant
rejection Therapy with protein A
Crystal structure of a Staphylococcus
aureus protein A domain complexed with
the Fab fragment of a human IgM antibody
Structural basis for recognition of B-cell
receptors and superantigen activity Marc Graille et al Proc Natl Acad Sci U S A
2000 May 9 97(10) 5399ndash5404 PMCID PMC25840
Immunology
Acute and chronic transplant
rejection Therapy with protein A
Staphylococcus aureus Protein A Promotes
Immune Suppression
Scott D Kobayashi
Frank R DeLeo
Acute and chronic transplant
rejection Therapy with protein A
SpA binds the Fc region of antibody and the Fabregions of the B-cell receptor processes that are known to block opsonophagocytosis and cause B-cell death in vitro In a recent study Falugi et al [F Falugi H K Kim D M Missiakas and O Schneewind mBio4(5)e00575-13 2013] showed that vaccination with spa mutant S aureus strains lacking antibody Fc-andor Fab-binding capacity protects against subsequent challenge with the USA300 epidemic strain The findings provide strong support for the idea that SpA promotes S aureus immune evasion in vivo and form the foundation for a new approach in our efforts to develop a vaccine that prevents severe S aureusinfections
Acute and chronic transplant
rejection Therapy with protein A
Humoral immunity
Developed through an earlier primary exposure that primed specific immunity to the nonself antigen a transplant recipient can have specific antibody crossreacting with the donor tissue upon the transplant event a secondary exposure This is typical after earlier mismatching among ABO blood types during blood transfusion At this secondary exposure these crossreactive antibody molecules interact with aspects of innate immunitymdashsoluble immune proteins called complement and innate immune cells calledphagocytesmdashwhich inflames and destroys the transplanted tissue
Acute and chronic transplant
rejection Therapy with protein A
Antibody[edit]
Secreted by an activated B cell then
called plasma cell an antibody molecule is a
soluble immunoglobulin (Ig) whose basic unit is
shaped like the letter Y the two arms are theFab
regions while the single stalk is the Fc region
Each of the two tips of Fab region is the paratope
which binds a matching molecular sequence and
its 3D shape (conformation) altogether
called epitope within the target antigen
Acute and chronic transplant
rejection Therapy with protein A
Opsonization
The IgGs Fc region also enables opsonization by a phagocyte a process by which the Fc receptor on the phagocytemdashsuch as neutrophils in blood and macrophages in tissuesmdashbinds the antibody molecules FC stalk and the phagocyte exhibits enhanced uptake of the antigen attached to the antibody molecules Fab region
Acute and chronic transplant
rejection Therapy with protein A
Complement cascade
When the paratope of Ig class gamma (IgG) binds its matching epitope IgGs Fc region conformationally shifts and can host a complement protein initiating the complement cascade that terminates by punching a hole in a cell membrane With many holes so punched fluid rushes into the cell and ruptures it
Cell debris can be recognized as damage associated molecular patterns (DAMPs) by pattern recognition receptors (PRRs) such as Toll-like receptors (TLRs) on membranes ofphagocytes which thereupon secrete proinflammatory cytokines recruiting more phagocytes to traffic to the area by sensing the concentration gradient of the secreted cytokines (chemotaxis)
Acute and chronic transplant
rejection Therapy with protein A
Protein A is a
42 kDa surface protein originally found in the
cell wall of the bacterium Staphylococcus
aureus It is encoded by the spa gene and its
regulation is controlled by DNA topology
cellular osmolarity and a two-component
system called ArlS-ArlR It has found use in
biochemical research because of its ability to
bind immunoglobulins
Acute and chronic transplant
rejection Therapy with protein A
It is composed of five homologous Ig-binding domains that fold into a three-helix bundle Each domain is able to bind proteins from many mammalian species most notably IgGs It binds the heavy chain within the Fc region of most immunoglobulins and also within the Fab region in the case of the human VH3 family Through these interactions in serum where IgG molecules are bound in the wrong orientation (in relation to normal antibodyfunction) the bacteria disrupts opsonization and phagocytosis
Acute and chronic transplant
rejection Therapy with protein A
Purified (unconjugated) Thermo Scientific Pierce Recombinant Protein A is useful as the basis for preparing various kinds of probes or affinity media for detection or purification of rabbit and human antibodies especially IgGisotypes in immunoassays and antibody purification protocols
Features of Recombinant Protein A
bull Contains four Fc-binding domains per proteinbull Better than Protein G for polyclonal IgG from rabbit pig dog and cat serumbull Poorer than Protein G for mouse IgG1 human IgG3 and rat goat and cow antibodies
Acute and chronic transplant
rejection Therapy with protein A
Protein A is a cell wall component produced by several strains of Staphylococcus aureus that consists of a single polypeptide chain and contains little or no carbohydrate Recombinant Protein A is produced in E coli and functions essentially the same as native Protein A The Protein A molecule contains four high-affinity binding sites capable of interacting with the Fcregion from IgG of several species including human and rabbit Optimal binding occurs at pH 82 although binding is also effective at neutral or physiological conditions (pH 70 to 76)
Acute and chronic transplant
rejection Therapy with protein A
httpswwwlifetechnologiescomordercatalog
product21184
Acute and chronic transplant
rejection Therapy with protein A
httpstoolslifetechnologiescomcontentsfsm
anualsMAN0011438_Pierce_Ig_Bind_Protein
s_UGpdf
httpstoolslifetechnologiescomcontentsfsbr
ochuresTR0034-Ab-binding-proteinspdf
Acute and chronic transplant
rejection Therapy with protein A
Crystal structure of a Staphylococcus
aureus protein A domain complexed with
the Fab fragment of a human IgM antibody
Structural basis for recognition of B-cell
receptors and superantigen activity Marc Graille et al Proc Natl Acad Sci U S A
2000 May 9 97(10) 5399ndash5404 PMCID PMC25840
Immunology
Acute and chronic transplant
rejection Therapy with protein A
Staphylococcus aureus Protein A Promotes
Immune Suppression
Scott D Kobayashi
Frank R DeLeo
Acute and chronic transplant
rejection Therapy with protein A
SpA binds the Fc region of antibody and the Fabregions of the B-cell receptor processes that are known to block opsonophagocytosis and cause B-cell death in vitro In a recent study Falugi et al [F Falugi H K Kim D M Missiakas and O Schneewind mBio4(5)e00575-13 2013] showed that vaccination with spa mutant S aureus strains lacking antibody Fc-andor Fab-binding capacity protects against subsequent challenge with the USA300 epidemic strain The findings provide strong support for the idea that SpA promotes S aureus immune evasion in vivo and form the foundation for a new approach in our efforts to develop a vaccine that prevents severe S aureusinfections
Acute and chronic transplant
rejection Therapy with protein A
Antibody[edit]
Secreted by an activated B cell then
called plasma cell an antibody molecule is a
soluble immunoglobulin (Ig) whose basic unit is
shaped like the letter Y the two arms are theFab
regions while the single stalk is the Fc region
Each of the two tips of Fab region is the paratope
which binds a matching molecular sequence and
its 3D shape (conformation) altogether
called epitope within the target antigen
Acute and chronic transplant
rejection Therapy with protein A
Opsonization
The IgGs Fc region also enables opsonization by a phagocyte a process by which the Fc receptor on the phagocytemdashsuch as neutrophils in blood and macrophages in tissuesmdashbinds the antibody molecules FC stalk and the phagocyte exhibits enhanced uptake of the antigen attached to the antibody molecules Fab region
Acute and chronic transplant
rejection Therapy with protein A
Complement cascade
When the paratope of Ig class gamma (IgG) binds its matching epitope IgGs Fc region conformationally shifts and can host a complement protein initiating the complement cascade that terminates by punching a hole in a cell membrane With many holes so punched fluid rushes into the cell and ruptures it
Cell debris can be recognized as damage associated molecular patterns (DAMPs) by pattern recognition receptors (PRRs) such as Toll-like receptors (TLRs) on membranes ofphagocytes which thereupon secrete proinflammatory cytokines recruiting more phagocytes to traffic to the area by sensing the concentration gradient of the secreted cytokines (chemotaxis)
Acute and chronic transplant
rejection Therapy with protein A
Protein A is a
42 kDa surface protein originally found in the
cell wall of the bacterium Staphylococcus
aureus It is encoded by the spa gene and its
regulation is controlled by DNA topology
cellular osmolarity and a two-component
system called ArlS-ArlR It has found use in
biochemical research because of its ability to
bind immunoglobulins
Acute and chronic transplant
rejection Therapy with protein A
It is composed of five homologous Ig-binding domains that fold into a three-helix bundle Each domain is able to bind proteins from many mammalian species most notably IgGs It binds the heavy chain within the Fc region of most immunoglobulins and also within the Fab region in the case of the human VH3 family Through these interactions in serum where IgG molecules are bound in the wrong orientation (in relation to normal antibodyfunction) the bacteria disrupts opsonization and phagocytosis
Acute and chronic transplant
rejection Therapy with protein A
Purified (unconjugated) Thermo Scientific Pierce Recombinant Protein A is useful as the basis for preparing various kinds of probes or affinity media for detection or purification of rabbit and human antibodies especially IgGisotypes in immunoassays and antibody purification protocols
Features of Recombinant Protein A
bull Contains four Fc-binding domains per proteinbull Better than Protein G for polyclonal IgG from rabbit pig dog and cat serumbull Poorer than Protein G for mouse IgG1 human IgG3 and rat goat and cow antibodies
Acute and chronic transplant
rejection Therapy with protein A
Protein A is a cell wall component produced by several strains of Staphylococcus aureus that consists of a single polypeptide chain and contains little or no carbohydrate Recombinant Protein A is produced in E coli and functions essentially the same as native Protein A The Protein A molecule contains four high-affinity binding sites capable of interacting with the Fcregion from IgG of several species including human and rabbit Optimal binding occurs at pH 82 although binding is also effective at neutral or physiological conditions (pH 70 to 76)
Acute and chronic transplant
rejection Therapy with protein A
httpswwwlifetechnologiescomordercatalog
product21184
Acute and chronic transplant
rejection Therapy with protein A
httpstoolslifetechnologiescomcontentsfsm
anualsMAN0011438_Pierce_Ig_Bind_Protein
s_UGpdf
httpstoolslifetechnologiescomcontentsfsbr
ochuresTR0034-Ab-binding-proteinspdf
Acute and chronic transplant
rejection Therapy with protein A
Crystal structure of a Staphylococcus
aureus protein A domain complexed with
the Fab fragment of a human IgM antibody
Structural basis for recognition of B-cell
receptors and superantigen activity Marc Graille et al Proc Natl Acad Sci U S A
2000 May 9 97(10) 5399ndash5404 PMCID PMC25840
Immunology
Acute and chronic transplant
rejection Therapy with protein A
Staphylococcus aureus Protein A Promotes
Immune Suppression
Scott D Kobayashi
Frank R DeLeo
Acute and chronic transplant
rejection Therapy with protein A
SpA binds the Fc region of antibody and the Fabregions of the B-cell receptor processes that are known to block opsonophagocytosis and cause B-cell death in vitro In a recent study Falugi et al [F Falugi H K Kim D M Missiakas and O Schneewind mBio4(5)e00575-13 2013] showed that vaccination with spa mutant S aureus strains lacking antibody Fc-andor Fab-binding capacity protects against subsequent challenge with the USA300 epidemic strain The findings provide strong support for the idea that SpA promotes S aureus immune evasion in vivo and form the foundation for a new approach in our efforts to develop a vaccine that prevents severe S aureusinfections
Acute and chronic transplant
rejection Therapy with protein A
Opsonization
The IgGs Fc region also enables opsonization by a phagocyte a process by which the Fc receptor on the phagocytemdashsuch as neutrophils in blood and macrophages in tissuesmdashbinds the antibody molecules FC stalk and the phagocyte exhibits enhanced uptake of the antigen attached to the antibody molecules Fab region
Acute and chronic transplant
rejection Therapy with protein A
Complement cascade
When the paratope of Ig class gamma (IgG) binds its matching epitope IgGs Fc region conformationally shifts and can host a complement protein initiating the complement cascade that terminates by punching a hole in a cell membrane With many holes so punched fluid rushes into the cell and ruptures it
Cell debris can be recognized as damage associated molecular patterns (DAMPs) by pattern recognition receptors (PRRs) such as Toll-like receptors (TLRs) on membranes ofphagocytes which thereupon secrete proinflammatory cytokines recruiting more phagocytes to traffic to the area by sensing the concentration gradient of the secreted cytokines (chemotaxis)
Acute and chronic transplant
rejection Therapy with protein A
Protein A is a
42 kDa surface protein originally found in the
cell wall of the bacterium Staphylococcus
aureus It is encoded by the spa gene and its
regulation is controlled by DNA topology
cellular osmolarity and a two-component
system called ArlS-ArlR It has found use in
biochemical research because of its ability to
bind immunoglobulins
Acute and chronic transplant
rejection Therapy with protein A
It is composed of five homologous Ig-binding domains that fold into a three-helix bundle Each domain is able to bind proteins from many mammalian species most notably IgGs It binds the heavy chain within the Fc region of most immunoglobulins and also within the Fab region in the case of the human VH3 family Through these interactions in serum where IgG molecules are bound in the wrong orientation (in relation to normal antibodyfunction) the bacteria disrupts opsonization and phagocytosis
Acute and chronic transplant
rejection Therapy with protein A
Purified (unconjugated) Thermo Scientific Pierce Recombinant Protein A is useful as the basis for preparing various kinds of probes or affinity media for detection or purification of rabbit and human antibodies especially IgGisotypes in immunoassays and antibody purification protocols
Features of Recombinant Protein A
bull Contains four Fc-binding domains per proteinbull Better than Protein G for polyclonal IgG from rabbit pig dog and cat serumbull Poorer than Protein G for mouse IgG1 human IgG3 and rat goat and cow antibodies
Acute and chronic transplant
rejection Therapy with protein A
Protein A is a cell wall component produced by several strains of Staphylococcus aureus that consists of a single polypeptide chain and contains little or no carbohydrate Recombinant Protein A is produced in E coli and functions essentially the same as native Protein A The Protein A molecule contains four high-affinity binding sites capable of interacting with the Fcregion from IgG of several species including human and rabbit Optimal binding occurs at pH 82 although binding is also effective at neutral or physiological conditions (pH 70 to 76)
Acute and chronic transplant
rejection Therapy with protein A
httpswwwlifetechnologiescomordercatalog
product21184
Acute and chronic transplant
rejection Therapy with protein A
httpstoolslifetechnologiescomcontentsfsm
anualsMAN0011438_Pierce_Ig_Bind_Protein
s_UGpdf
httpstoolslifetechnologiescomcontentsfsbr
ochuresTR0034-Ab-binding-proteinspdf
Acute and chronic transplant
rejection Therapy with protein A
Crystal structure of a Staphylococcus
aureus protein A domain complexed with
the Fab fragment of a human IgM antibody
Structural basis for recognition of B-cell
receptors and superantigen activity Marc Graille et al Proc Natl Acad Sci U S A
2000 May 9 97(10) 5399ndash5404 PMCID PMC25840
Immunology
Acute and chronic transplant
rejection Therapy with protein A
Staphylococcus aureus Protein A Promotes
Immune Suppression
Scott D Kobayashi
Frank R DeLeo
Acute and chronic transplant
rejection Therapy with protein A
SpA binds the Fc region of antibody and the Fabregions of the B-cell receptor processes that are known to block opsonophagocytosis and cause B-cell death in vitro In a recent study Falugi et al [F Falugi H K Kim D M Missiakas and O Schneewind mBio4(5)e00575-13 2013] showed that vaccination with spa mutant S aureus strains lacking antibody Fc-andor Fab-binding capacity protects against subsequent challenge with the USA300 epidemic strain The findings provide strong support for the idea that SpA promotes S aureus immune evasion in vivo and form the foundation for a new approach in our efforts to develop a vaccine that prevents severe S aureusinfections
Acute and chronic transplant
rejection Therapy with protein A
Complement cascade
When the paratope of Ig class gamma (IgG) binds its matching epitope IgGs Fc region conformationally shifts and can host a complement protein initiating the complement cascade that terminates by punching a hole in a cell membrane With many holes so punched fluid rushes into the cell and ruptures it
Cell debris can be recognized as damage associated molecular patterns (DAMPs) by pattern recognition receptors (PRRs) such as Toll-like receptors (TLRs) on membranes ofphagocytes which thereupon secrete proinflammatory cytokines recruiting more phagocytes to traffic to the area by sensing the concentration gradient of the secreted cytokines (chemotaxis)
Acute and chronic transplant
rejection Therapy with protein A
Protein A is a
42 kDa surface protein originally found in the
cell wall of the bacterium Staphylococcus
aureus It is encoded by the spa gene and its
regulation is controlled by DNA topology
cellular osmolarity and a two-component
system called ArlS-ArlR It has found use in
biochemical research because of its ability to
bind immunoglobulins
Acute and chronic transplant
rejection Therapy with protein A
It is composed of five homologous Ig-binding domains that fold into a three-helix bundle Each domain is able to bind proteins from many mammalian species most notably IgGs It binds the heavy chain within the Fc region of most immunoglobulins and also within the Fab region in the case of the human VH3 family Through these interactions in serum where IgG molecules are bound in the wrong orientation (in relation to normal antibodyfunction) the bacteria disrupts opsonization and phagocytosis
Acute and chronic transplant
rejection Therapy with protein A
Purified (unconjugated) Thermo Scientific Pierce Recombinant Protein A is useful as the basis for preparing various kinds of probes or affinity media for detection or purification of rabbit and human antibodies especially IgGisotypes in immunoassays and antibody purification protocols
Features of Recombinant Protein A
bull Contains four Fc-binding domains per proteinbull Better than Protein G for polyclonal IgG from rabbit pig dog and cat serumbull Poorer than Protein G for mouse IgG1 human IgG3 and rat goat and cow antibodies
Acute and chronic transplant
rejection Therapy with protein A
Protein A is a cell wall component produced by several strains of Staphylococcus aureus that consists of a single polypeptide chain and contains little or no carbohydrate Recombinant Protein A is produced in E coli and functions essentially the same as native Protein A The Protein A molecule contains four high-affinity binding sites capable of interacting with the Fcregion from IgG of several species including human and rabbit Optimal binding occurs at pH 82 although binding is also effective at neutral or physiological conditions (pH 70 to 76)
Acute and chronic transplant
rejection Therapy with protein A
httpswwwlifetechnologiescomordercatalog
product21184
Acute and chronic transplant
rejection Therapy with protein A
httpstoolslifetechnologiescomcontentsfsm
anualsMAN0011438_Pierce_Ig_Bind_Protein
s_UGpdf
httpstoolslifetechnologiescomcontentsfsbr
ochuresTR0034-Ab-binding-proteinspdf
Acute and chronic transplant
rejection Therapy with protein A
Crystal structure of a Staphylococcus
aureus protein A domain complexed with
the Fab fragment of a human IgM antibody
Structural basis for recognition of B-cell
receptors and superantigen activity Marc Graille et al Proc Natl Acad Sci U S A
2000 May 9 97(10) 5399ndash5404 PMCID PMC25840
Immunology
Acute and chronic transplant
rejection Therapy with protein A
Staphylococcus aureus Protein A Promotes
Immune Suppression
Scott D Kobayashi
Frank R DeLeo
Acute and chronic transplant
rejection Therapy with protein A
SpA binds the Fc region of antibody and the Fabregions of the B-cell receptor processes that are known to block opsonophagocytosis and cause B-cell death in vitro In a recent study Falugi et al [F Falugi H K Kim D M Missiakas and O Schneewind mBio4(5)e00575-13 2013] showed that vaccination with spa mutant S aureus strains lacking antibody Fc-andor Fab-binding capacity protects against subsequent challenge with the USA300 epidemic strain The findings provide strong support for the idea that SpA promotes S aureus immune evasion in vivo and form the foundation for a new approach in our efforts to develop a vaccine that prevents severe S aureusinfections
Acute and chronic transplant
rejection Therapy with protein A
Protein A is a
42 kDa surface protein originally found in the
cell wall of the bacterium Staphylococcus
aureus It is encoded by the spa gene and its
regulation is controlled by DNA topology
cellular osmolarity and a two-component
system called ArlS-ArlR It has found use in
biochemical research because of its ability to
bind immunoglobulins
Acute and chronic transplant
rejection Therapy with protein A
It is composed of five homologous Ig-binding domains that fold into a three-helix bundle Each domain is able to bind proteins from many mammalian species most notably IgGs It binds the heavy chain within the Fc region of most immunoglobulins and also within the Fab region in the case of the human VH3 family Through these interactions in serum where IgG molecules are bound in the wrong orientation (in relation to normal antibodyfunction) the bacteria disrupts opsonization and phagocytosis
Acute and chronic transplant
rejection Therapy with protein A
Purified (unconjugated) Thermo Scientific Pierce Recombinant Protein A is useful as the basis for preparing various kinds of probes or affinity media for detection or purification of rabbit and human antibodies especially IgGisotypes in immunoassays and antibody purification protocols
Features of Recombinant Protein A
bull Contains four Fc-binding domains per proteinbull Better than Protein G for polyclonal IgG from rabbit pig dog and cat serumbull Poorer than Protein G for mouse IgG1 human IgG3 and rat goat and cow antibodies
Acute and chronic transplant
rejection Therapy with protein A
Protein A is a cell wall component produced by several strains of Staphylococcus aureus that consists of a single polypeptide chain and contains little or no carbohydrate Recombinant Protein A is produced in E coli and functions essentially the same as native Protein A The Protein A molecule contains four high-affinity binding sites capable of interacting with the Fcregion from IgG of several species including human and rabbit Optimal binding occurs at pH 82 although binding is also effective at neutral or physiological conditions (pH 70 to 76)
Acute and chronic transplant
rejection Therapy with protein A
httpswwwlifetechnologiescomordercatalog
product21184
Acute and chronic transplant
rejection Therapy with protein A
httpstoolslifetechnologiescomcontentsfsm
anualsMAN0011438_Pierce_Ig_Bind_Protein
s_UGpdf
httpstoolslifetechnologiescomcontentsfsbr
ochuresTR0034-Ab-binding-proteinspdf
Acute and chronic transplant
rejection Therapy with protein A
Crystal structure of a Staphylococcus
aureus protein A domain complexed with
the Fab fragment of a human IgM antibody
Structural basis for recognition of B-cell
receptors and superantigen activity Marc Graille et al Proc Natl Acad Sci U S A
2000 May 9 97(10) 5399ndash5404 PMCID PMC25840
Immunology
Acute and chronic transplant
rejection Therapy with protein A
Staphylococcus aureus Protein A Promotes
Immune Suppression
Scott D Kobayashi
Frank R DeLeo
Acute and chronic transplant
rejection Therapy with protein A
SpA binds the Fc region of antibody and the Fabregions of the B-cell receptor processes that are known to block opsonophagocytosis and cause B-cell death in vitro In a recent study Falugi et al [F Falugi H K Kim D M Missiakas and O Schneewind mBio4(5)e00575-13 2013] showed that vaccination with spa mutant S aureus strains lacking antibody Fc-andor Fab-binding capacity protects against subsequent challenge with the USA300 epidemic strain The findings provide strong support for the idea that SpA promotes S aureus immune evasion in vivo and form the foundation for a new approach in our efforts to develop a vaccine that prevents severe S aureusinfections
Acute and chronic transplant
rejection Therapy with protein A
It is composed of five homologous Ig-binding domains that fold into a three-helix bundle Each domain is able to bind proteins from many mammalian species most notably IgGs It binds the heavy chain within the Fc region of most immunoglobulins and also within the Fab region in the case of the human VH3 family Through these interactions in serum where IgG molecules are bound in the wrong orientation (in relation to normal antibodyfunction) the bacteria disrupts opsonization and phagocytosis
Acute and chronic transplant
rejection Therapy with protein A
Purified (unconjugated) Thermo Scientific Pierce Recombinant Protein A is useful as the basis for preparing various kinds of probes or affinity media for detection or purification of rabbit and human antibodies especially IgGisotypes in immunoassays and antibody purification protocols
Features of Recombinant Protein A
bull Contains four Fc-binding domains per proteinbull Better than Protein G for polyclonal IgG from rabbit pig dog and cat serumbull Poorer than Protein G for mouse IgG1 human IgG3 and rat goat and cow antibodies
Acute and chronic transplant
rejection Therapy with protein A
Protein A is a cell wall component produced by several strains of Staphylococcus aureus that consists of a single polypeptide chain and contains little or no carbohydrate Recombinant Protein A is produced in E coli and functions essentially the same as native Protein A The Protein A molecule contains four high-affinity binding sites capable of interacting with the Fcregion from IgG of several species including human and rabbit Optimal binding occurs at pH 82 although binding is also effective at neutral or physiological conditions (pH 70 to 76)
Acute and chronic transplant
rejection Therapy with protein A
httpswwwlifetechnologiescomordercatalog
product21184
Acute and chronic transplant
rejection Therapy with protein A
httpstoolslifetechnologiescomcontentsfsm
anualsMAN0011438_Pierce_Ig_Bind_Protein
s_UGpdf
httpstoolslifetechnologiescomcontentsfsbr
ochuresTR0034-Ab-binding-proteinspdf
Acute and chronic transplant
rejection Therapy with protein A
Crystal structure of a Staphylococcus
aureus protein A domain complexed with
the Fab fragment of a human IgM antibody
Structural basis for recognition of B-cell
receptors and superantigen activity Marc Graille et al Proc Natl Acad Sci U S A
2000 May 9 97(10) 5399ndash5404 PMCID PMC25840
Immunology
Acute and chronic transplant
rejection Therapy with protein A
Staphylococcus aureus Protein A Promotes
Immune Suppression
Scott D Kobayashi
Frank R DeLeo
Acute and chronic transplant
rejection Therapy with protein A
SpA binds the Fc region of antibody and the Fabregions of the B-cell receptor processes that are known to block opsonophagocytosis and cause B-cell death in vitro In a recent study Falugi et al [F Falugi H K Kim D M Missiakas and O Schneewind mBio4(5)e00575-13 2013] showed that vaccination with spa mutant S aureus strains lacking antibody Fc-andor Fab-binding capacity protects against subsequent challenge with the USA300 epidemic strain The findings provide strong support for the idea that SpA promotes S aureus immune evasion in vivo and form the foundation for a new approach in our efforts to develop a vaccine that prevents severe S aureusinfections
Acute and chronic transplant
rejection Therapy with protein A
Purified (unconjugated) Thermo Scientific Pierce Recombinant Protein A is useful as the basis for preparing various kinds of probes or affinity media for detection or purification of rabbit and human antibodies especially IgGisotypes in immunoassays and antibody purification protocols
Features of Recombinant Protein A
bull Contains four Fc-binding domains per proteinbull Better than Protein G for polyclonal IgG from rabbit pig dog and cat serumbull Poorer than Protein G for mouse IgG1 human IgG3 and rat goat and cow antibodies
Acute and chronic transplant
rejection Therapy with protein A
Protein A is a cell wall component produced by several strains of Staphylococcus aureus that consists of a single polypeptide chain and contains little or no carbohydrate Recombinant Protein A is produced in E coli and functions essentially the same as native Protein A The Protein A molecule contains four high-affinity binding sites capable of interacting with the Fcregion from IgG of several species including human and rabbit Optimal binding occurs at pH 82 although binding is also effective at neutral or physiological conditions (pH 70 to 76)
Acute and chronic transplant
rejection Therapy with protein A
httpswwwlifetechnologiescomordercatalog
product21184
Acute and chronic transplant
rejection Therapy with protein A
httpstoolslifetechnologiescomcontentsfsm
anualsMAN0011438_Pierce_Ig_Bind_Protein
s_UGpdf
httpstoolslifetechnologiescomcontentsfsbr
ochuresTR0034-Ab-binding-proteinspdf
Acute and chronic transplant
rejection Therapy with protein A
Crystal structure of a Staphylococcus
aureus protein A domain complexed with
the Fab fragment of a human IgM antibody
Structural basis for recognition of B-cell
receptors and superantigen activity Marc Graille et al Proc Natl Acad Sci U S A
2000 May 9 97(10) 5399ndash5404 PMCID PMC25840
Immunology
Acute and chronic transplant
rejection Therapy with protein A
Staphylococcus aureus Protein A Promotes
Immune Suppression
Scott D Kobayashi
Frank R DeLeo
Acute and chronic transplant
rejection Therapy with protein A
SpA binds the Fc region of antibody and the Fabregions of the B-cell receptor processes that are known to block opsonophagocytosis and cause B-cell death in vitro In a recent study Falugi et al [F Falugi H K Kim D M Missiakas and O Schneewind mBio4(5)e00575-13 2013] showed that vaccination with spa mutant S aureus strains lacking antibody Fc-andor Fab-binding capacity protects against subsequent challenge with the USA300 epidemic strain The findings provide strong support for the idea that SpA promotes S aureus immune evasion in vivo and form the foundation for a new approach in our efforts to develop a vaccine that prevents severe S aureusinfections
Acute and chronic transplant
rejection Therapy with protein A
Protein A is a cell wall component produced by several strains of Staphylococcus aureus that consists of a single polypeptide chain and contains little or no carbohydrate Recombinant Protein A is produced in E coli and functions essentially the same as native Protein A The Protein A molecule contains four high-affinity binding sites capable of interacting with the Fcregion from IgG of several species including human and rabbit Optimal binding occurs at pH 82 although binding is also effective at neutral or physiological conditions (pH 70 to 76)
Acute and chronic transplant
rejection Therapy with protein A
httpswwwlifetechnologiescomordercatalog
product21184
Acute and chronic transplant
rejection Therapy with protein A
httpstoolslifetechnologiescomcontentsfsm
anualsMAN0011438_Pierce_Ig_Bind_Protein
s_UGpdf
httpstoolslifetechnologiescomcontentsfsbr
ochuresTR0034-Ab-binding-proteinspdf
Acute and chronic transplant
rejection Therapy with protein A
Crystal structure of a Staphylococcus
aureus protein A domain complexed with
the Fab fragment of a human IgM antibody
Structural basis for recognition of B-cell
receptors and superantigen activity Marc Graille et al Proc Natl Acad Sci U S A
2000 May 9 97(10) 5399ndash5404 PMCID PMC25840
Immunology
Acute and chronic transplant
rejection Therapy with protein A
Staphylococcus aureus Protein A Promotes
Immune Suppression
Scott D Kobayashi
Frank R DeLeo
Acute and chronic transplant
rejection Therapy with protein A
SpA binds the Fc region of antibody and the Fabregions of the B-cell receptor processes that are known to block opsonophagocytosis and cause B-cell death in vitro In a recent study Falugi et al [F Falugi H K Kim D M Missiakas and O Schneewind mBio4(5)e00575-13 2013] showed that vaccination with spa mutant S aureus strains lacking antibody Fc-andor Fab-binding capacity protects against subsequent challenge with the USA300 epidemic strain The findings provide strong support for the idea that SpA promotes S aureus immune evasion in vivo and form the foundation for a new approach in our efforts to develop a vaccine that prevents severe S aureusinfections
Acute and chronic transplant
rejection Therapy with protein A
httpswwwlifetechnologiescomordercatalog
product21184
Acute and chronic transplant
rejection Therapy with protein A
httpstoolslifetechnologiescomcontentsfsm
anualsMAN0011438_Pierce_Ig_Bind_Protein
s_UGpdf
httpstoolslifetechnologiescomcontentsfsbr
ochuresTR0034-Ab-binding-proteinspdf
Acute and chronic transplant
rejection Therapy with protein A
Crystal structure of a Staphylococcus
aureus protein A domain complexed with
the Fab fragment of a human IgM antibody
Structural basis for recognition of B-cell
receptors and superantigen activity Marc Graille et al Proc Natl Acad Sci U S A
2000 May 9 97(10) 5399ndash5404 PMCID PMC25840
Immunology
Acute and chronic transplant
rejection Therapy with protein A
Staphylococcus aureus Protein A Promotes
Immune Suppression
Scott D Kobayashi
Frank R DeLeo
Acute and chronic transplant
rejection Therapy with protein A
SpA binds the Fc region of antibody and the Fabregions of the B-cell receptor processes that are known to block opsonophagocytosis and cause B-cell death in vitro In a recent study Falugi et al [F Falugi H K Kim D M Missiakas and O Schneewind mBio4(5)e00575-13 2013] showed that vaccination with spa mutant S aureus strains lacking antibody Fc-andor Fab-binding capacity protects against subsequent challenge with the USA300 epidemic strain The findings provide strong support for the idea that SpA promotes S aureus immune evasion in vivo and form the foundation for a new approach in our efforts to develop a vaccine that prevents severe S aureusinfections
Acute and chronic transplant
rejection Therapy with protein A
httpstoolslifetechnologiescomcontentsfsm
anualsMAN0011438_Pierce_Ig_Bind_Protein
s_UGpdf
httpstoolslifetechnologiescomcontentsfsbr
ochuresTR0034-Ab-binding-proteinspdf
Acute and chronic transplant
rejection Therapy with protein A
Crystal structure of a Staphylococcus
aureus protein A domain complexed with
the Fab fragment of a human IgM antibody
Structural basis for recognition of B-cell
receptors and superantigen activity Marc Graille et al Proc Natl Acad Sci U S A
2000 May 9 97(10) 5399ndash5404 PMCID PMC25840
Immunology
Acute and chronic transplant
rejection Therapy with protein A
Staphylococcus aureus Protein A Promotes
Immune Suppression
Scott D Kobayashi
Frank R DeLeo
Acute and chronic transplant
rejection Therapy with protein A
SpA binds the Fc region of antibody and the Fabregions of the B-cell receptor processes that are known to block opsonophagocytosis and cause B-cell death in vitro In a recent study Falugi et al [F Falugi H K Kim D M Missiakas and O Schneewind mBio4(5)e00575-13 2013] showed that vaccination with spa mutant S aureus strains lacking antibody Fc-andor Fab-binding capacity protects against subsequent challenge with the USA300 epidemic strain The findings provide strong support for the idea that SpA promotes S aureus immune evasion in vivo and form the foundation for a new approach in our efforts to develop a vaccine that prevents severe S aureusinfections
Acute and chronic transplant
rejection Therapy with protein A
Crystal structure of a Staphylococcus
aureus protein A domain complexed with
the Fab fragment of a human IgM antibody
Structural basis for recognition of B-cell
receptors and superantigen activity Marc Graille et al Proc Natl Acad Sci U S A
2000 May 9 97(10) 5399ndash5404 PMCID PMC25840
Immunology
Acute and chronic transplant
rejection Therapy with protein A
Staphylococcus aureus Protein A Promotes
Immune Suppression
Scott D Kobayashi
Frank R DeLeo
Acute and chronic transplant
rejection Therapy with protein A
SpA binds the Fc region of antibody and the Fabregions of the B-cell receptor processes that are known to block opsonophagocytosis and cause B-cell death in vitro In a recent study Falugi et al [F Falugi H K Kim D M Missiakas and O Schneewind mBio4(5)e00575-13 2013] showed that vaccination with spa mutant S aureus strains lacking antibody Fc-andor Fab-binding capacity protects against subsequent challenge with the USA300 epidemic strain The findings provide strong support for the idea that SpA promotes S aureus immune evasion in vivo and form the foundation for a new approach in our efforts to develop a vaccine that prevents severe S aureusinfections
Acute and chronic transplant
rejection Therapy with protein A
Staphylococcus aureus Protein A Promotes
Immune Suppression
Scott D Kobayashi
Frank R DeLeo
Acute and chronic transplant
rejection Therapy with protein A
SpA binds the Fc region of antibody and the Fabregions of the B-cell receptor processes that are known to block opsonophagocytosis and cause B-cell death in vitro In a recent study Falugi et al [F Falugi H K Kim D M Missiakas and O Schneewind mBio4(5)e00575-13 2013] showed that vaccination with spa mutant S aureus strains lacking antibody Fc-andor Fab-binding capacity protects against subsequent challenge with the USA300 epidemic strain The findings provide strong support for the idea that SpA promotes S aureus immune evasion in vivo and form the foundation for a new approach in our efforts to develop a vaccine that prevents severe S aureusinfections
Acute and chronic transplant
rejection Therapy with protein A
SpA binds the Fc region of antibody and the Fabregions of the B-cell receptor processes that are known to block opsonophagocytosis and cause B-cell death in vitro In a recent study Falugi et al [F Falugi H K Kim D M Missiakas and O Schneewind mBio4(5)e00575-13 2013] showed that vaccination with spa mutant S aureus strains lacking antibody Fc-andor Fab-binding capacity protects against subsequent challenge with the USA300 epidemic strain The findings provide strong support for the idea that SpA promotes S aureus immune evasion in vivo and form the foundation for a new approach in our efforts to develop a vaccine that prevents severe S aureusinfections