immuno hematology

IMMUNOHEMATOLOGY

( BLOOD BANKING )

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Definition of terms:ReviewA. Absorption

= removal of an unwanted antibody from a serum; often used interchangeably with adsorption.

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Adsorption= providing an Ab with its corresponding antigen under optimal conditions so that the Ab will attach to the Ag. It is thus take out of the serum . Often used interchangeably with absorption.

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Agglutination= the clumping together of RBC or any particulate matter resulting from interaction of Ab & its corresponding Ag.

Agglutinin = an Ab that agglutinates cell.

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Agglutinogen= a substance that stimulates the production of an agglutinin, thereby, acting as an Ag.Antibody = a protein subs. developed in response to, & interacting specifically w/ an Ag. In Blood banking it is found serum, from either a commercial manufacturer or a patient. It is secreted by plasma

cells.

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Apheresis= a method of blood collection in w/c whole blood is withdrawn, a desired component separated and retained, & the remainder of the blood returned to the donor.

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Avidity= is a measure of a functional affinity of an antiserum for the whole antigen.

Elution= a process whereby cells that are coated with Ab are treated in such a manner as to disrupt the bonds bet. the Ag and Ab.

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Antigen= a substance that is recognized by the body as being foreign thus it can elicit immune response.

Fibrinolysin/ Plasmin= has the ability to dissolve fibrin.

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Fibrinolysis= dissolution of fibrin by fibrinolysin caused by the action of a proteolytic enzyme system that is continually active in the body but that is increased greatly by various stress stimuli.

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Hemolysin= an antibody that activates complement leading to cell lysis.

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Hemagglutination= agglutination of RBC or clumping of RBC.

Hemaaglutinin = a protein in blood serum that causes clumping of RBC; also present in the surface projection of some viruses.

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Hemolysis= disruption of the red cell membrane and the subsequent release of hemoglobin into the suspending medium or plasma.

Immunogen= any substance capable of stimulating an immune response.

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Immunogenecity

= a descriptive term indicating the ability of an antigen to stimulate an Ab response.

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Immunoglobulin= one of a family of closely related though not identical proteins that are capable of acting as antibodies. They are IgA, IgG, IgM, IgD, IgE

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IgG= is the main Ig in human serum

IgA= is the principal Ig in exocrine secretions such as saliva & tears.

IgM= is a globulin formed in almost every immune response during the early period of the reaction.

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IgD= may play a role in Ag recognition & the initiation of Ab synthesis.

IgE= is produced by the cell lining the intestinal & respiratory tract & is important in forming reagin.

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LECTIN= proteins that are present in plants ( usually seeds) w/c binds specially to carbohydrates determinants & agglutinate RBC through their cell surface oligosaccharide determinants.

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Phlebotomy= to take blood from a

person.

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Sensitization= a condition of being made sensitive to a specific substance( e.g. an Ag) after the initial exposure to the substance.

Transfusion = the injection of blood, a blood component, saline, or other fluids into the bloodstream.

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B. Allele= one of the two or more different genes that may occupy a specific locus on a chromosome.

Chromosomes= the structure w/n the nucleus that contain a linear thread of DNA, w/c transmits genetic information.

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Chromosome= Genes are arranged along the strand of DNA and constitute portions of DNA.

Codominant Gene= a pair of genes in w/c neither is dominant over the other, that they are both expressed.

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Dominant gene= a trait or characteristic that will be expressed in the offspring even though it is only carried on one of the homologous chromosomes.

Gene= a unit of inheritance w/n a chromosome.

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Genotype= an individual’s actual genetic make- up.

Heterozygous= processing different alleles at a given locus.

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Homozygous= possessing a pair of identical alleles.

Locus= the site of a gene on a chromosome.

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Meiosis= type of cell division of germ cells in w/c 2 successive divisions of the nucleus produce cell that contain half the no. of chromosomes present in somatic cells.

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Mitosis= type of cell division in w/c

each daughter cell contains the same number of chromosomes as the parent cell. All cell except sex cells undergo mitosis.

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Interphase= resting stage from cell division.

Prophase= coiling of DNA protein complex; chromosome duplication.

Metaphase= alignment of chromosome in equatorial plane of cell.

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Telophase= constriction develops at the times of equatorial plane of mother cells.

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Phenotype= the outward expression of genes (e.g a blood type). On blood cells, serologically demonstrate Ag’s constitute by the phenotype except those sugar sites that are determinant by transferases.

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Recessive gene= a gene that, in the presence of its dominant allele, does not express itself. Expression of recessive genes occurs when they are inherited in the homozygous genes or state.

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Zygote= the single fertilized cells formed by the union of two gametes.

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PHASES OF HEMOSTATIC MECHANISM

Vasoconstriction= is the initial response (neurogenic) to vessel damage.

Platelet aggregation= by its ability to clump & forms a temporary plug to the blood flow outlet.

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Coagulation= by the activation of plasma factors forming a permanent plug w/c promotes vessel healing.

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DNA AND RNA

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DNA= is composed of a pentose

sugar lacking in oxygen known as Deoxyribose, nitrogenous bases (guanine, cytosine, thymine & adenine) & a phosphate fastened together forming a double helix structure.

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RNA= is likewise composed of a pentose sugar, ribose, nitrogenous bases ( guanine, cytosine, thymine replace by uracil & adenine) & forms a single coiled structure.= it is also contains phosphate bonds or groups.

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Types or forms of RNA:1.rRNA = 80%2.tRNA = 15% - adaptor in

protein synthesis; identifying/ picking up of codon.

3.mRNA = 5% - template of CHON synthesis.

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Processes:1. Replication/ duplication

= model: Watson- Cricka. semi- conservativeb. conservativeMajor enzymes:

a. DNA polymeraseb. DNA ligase

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2. Transcriptionsteps:a. binding of RNA polymerase

( major enzyme for transcription) to DNA.b. initiation of polymerizationc. chain elongationd. termination of synthesis

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3. Translation ( Protein synthesis stage)1. hydrogen bond between complementary.2. covalent bond between sugar and phosphate3. interaction between sugar and phosphate.

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4. Interaction between DNA and histones.

5. Van der Waals of hydrophobic forces

…….stabilizing factor of RNA

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Differences:DNA RNA

Double stranded single stranded

Bigger smallerDeoxyribose (sugar) ribose (sugar)Tymine- adenine uracil- adenine

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PROTEIN SYNTHESISINITIATION

=involves a collection of processes w/c starts in the nucleotides sequence AGGAGGU; initiation ends in the attachment of 1st tRNA carrying amino acid.

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ELONGATION= starts in the attachment of a 2nd tRNA w/ amino acid to a codon of mRNA through the ribosomal subunits; the amino acid binds w/ the first one & this processes will continue until a suitable or the dictated polypepetide chain is reached.

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TERMINATION= after repetition of the above process, elongation will stop at the last tRNA attached by the mRNA as dictated by the transcripted strand.

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IMMUNOHEMATOLOGY

= refers to immunologic reactions involving red blood cells and blood components.= immunologic properties and reactions of all blood components and con constituents.

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APPLICATION1. Safe blood transfusion and

organ transplantation.2. Understanding the

pathogenesis, diagnosis and prevention of Rh immunization associated with pregnancy.

3. Resolution of disputed parentage problem.

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BLOOD BANKING

= embraces with the collection, processing, preparation and distribution of blood components and its derivatives.

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Review on BLOODBLOOD

= nutritive fluid that circulates all throughout the body.= produce and mature in the bone marrow, released into the bloodstream where they play important functions.

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• An adult human has about 4–6 liters of blood circulating in the body. Among other things, blood transports oxygen to various parts of the body.

• Blood consists of several types of cells floating around in a fluid called plasma.

The red blood cells contain hemoglobin, a protein that binds oxygen. Red blood cells transport oxygen to, and remove carbon dioxide from, the body tissues.

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• The platelets help the blood to clot, if you get a wound for example.

The plasma contains salts and various kinds of proteins.

06-15-12

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Functions:1. O2 transport and facilitates

CO2 excretion.2. Transport of hormone from

the organ of production.3. Excretion of waste product

from cellular metabolism.

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4. Regulation of body temperature.

5. Regulation of acid- base balance.

6. Involved body defense mechanism.

7. Blood coagulation and hemostasis.

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CompositionA. Formed elements= 45% of

total or cellular elements

volume

1. RBC 2. WBC3. Platelets

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B. Fluid portion= 55% of total volume

PLASMA OR SERUM1. H20 = 91.5%2. electrolytes3. proteins

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Characteristics or Properties1. Fluid form but coagulates in

vitro2. Red in color due to Hb.3. Thick and viscous (3.5-4.5u)4. Slightly alkaline

( 7.35- 7.45 )

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5. SG = 1.055 (1.045- 1.065)

6. Comprises 7.8% of total body weight or 75-85ml/kg body weight

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RBC membrane= blood group antigens found.= consist of bilipid layer formed

by orderly arrangement of phospholipids.

= embedded in the lipid bilayers are protein and other complex molecules such as carbohydrates.

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Proteins are:1. Integral CHON = outside

(glycophorin)2. Peripheral CHON=

cytoplasmic(e.g. spectrin)

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Biochemical Composition of Cell Membrane

a. 52% CHON = for enzymatic reaction

= structural stabilityb. 89% CHO= cell specifityc. 40% lipids

= selective permeability= rigidity= fluidity

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Factors affecting red cell survival

1. Normal chemical composition.

2. Structural arrangement of components.

3. Interaction of erythrocyte membrane.

4. Deformability ( fluidity)

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4. Deformability= due to decrease ATP level, increase calcium deposition in cell membrane (increase rigidity)

5. Permeability= electrolyte and water balance.

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Metabolic pathways that takes place in RBC

1. Anaerobic glycolysis= generates 90% ATP

2. Pentose phosphate pathway= generates 10% ATP

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2. PPPDeficient PPP reduce glutathione

(insufficient to neutralize intracellular oxidants)

result in globin precipitation as aggregate ( Heinz bodies)

cell membrane damage & cell destruction

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3. Methemoglobin reductase pathway

a. NADH methemoglobin reductase*

b. NADPH methemoglobin reductase

Absence of methemoglobin reductase= accumulation of methemoglobin (Abnormal Hgb) due to conversion of Fe++ in Hb to Fe+++ w/c alters O2 transport

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4. Luebering- Rapoport Shunt Pathway

= produce 2,3- diphosphoglycerate

( 2,3- DPG)Increase 2,3- DPG level=

decrease Hb-O2 affinity w/c is good in oxygen transport.

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Assignment:1. What is gene mutation?2. What are the types of gene

mutation? Explain comprehensively, give example if necessary.

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HISTORICAL BACKGROUD• 1492 - Pope Innocent VIII, in

Rome, had an apoplectic stroke; became weak and went into a coma. His physician advised a Blood transfusion as a therapeutic measure for the Pope's illness. Employing crude methods, the Pope did not benefit and died by the end of that year.

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1615 - Andreas Libavius described his technique of Blood transfusion. It was unfortunately not adequately publicized.1665 - The first  Blood transfusions of

record take place. Animal experiments conducted by Richard Lower, an Oxford physician started as dog-to-dog experiments and proceeded to animal-to-human over the next two years. Dogs were kept alive by the transfusion of Blood from other dogs.

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1667 - Jean-Baptiste Denis in France

= successful transfusions from sheep to humans.

1678= Transfusion from animals to humans, having been tried in many different ways, was deemed to be unsuccessful, and was subsequently outlawed by the Paris Society of Physicians because of reactions, many resulting in death.

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1818 - James Blundell, a British obstetrician, performed the first successful transfusion of human Blood to a patient for the treatment of postpartum hemorrhage. Using the patient's husband as a donor, he extracted a small amount of Blood from the husband's arm and, using a syringe, he successfully transfused the wife.

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• 1840 - In London England, Samuel Armstrong Lane, aided by consultant Dr. Blundell, performed the first successful whole Blood transfusion to treat hemophilia.

• 1867 - English surgeon Joseph Lister utilized antiseptics to control infection during Blood transfusions.

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Fr. Of BB

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.1901 - Karl Landsteiner, an Austrian physician, and the most important individual in the field of Blood transfusion, documented the first three human Blood groups (based on substances present on the red Blood cells), A, B and O.= Father of Immunohematology.

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1902 - A fourth main Blood type, AB was found by A. Decastrello and A. Sturli.

1907 - Hektoen suggested that the safety of transfusion might be improved by cross-matching Blood between donors and patients to exclude incompatible mixtures.

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Reuben Ottenberg = performed the first Blood transfusion using Blood typing and cross-matching.

= also observed the 'Mendelian inheritance' of Blood groups and recognized the “universal” utility of group O donors.

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1908 Epstein & Ottenberg= how Ag’s produced= blood groups are

inherited.1909 Bateson

= blood groups are acquired.

1914 Hustin= Sodium Citrate as

anticoagulant.

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1915 Liwisohn= determined the

minimum amount of citrate to be used.

1916 Rous &Turner= Citrate Dextrose

1934 Berstein= supported the

postulate of Ottenberg & Epstein.

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1939- 40 Landsteiner – WeinerLivine & Stetson

= discovered Rh and HDN

1943 Loutit & Turner= introduced ACD

1948-49 Morgan, Watkins & Kabat

= discovered ABO structure.

1957 Gibson= introduced CPD.

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Immunohematology