What is immunohematology? Serologic, genetic, biochemical, and molecular study of antigens associated with membrane structures on the cellular constituents of blood, as well as the immunologic properties and reactions of blood component (Henry, 2011) Study of blood related antigens and antibodies as applied to situations in blood bank and the transfusion service (Whitlock, 2010)

Historical Timeline 1492 Pope Innocent VII First recorded blood transfusion in history 1667 Jean Baptiste Denis: first recorded animal-to-human blood transfusion (calf blood) Richard Lower: sheeps blood 1795 Philip Syng Physick: unconfirmed first human-to-human transfusion 1816 John Henry Leacock: On the Transfusion of Blood in Extreme Cases of Hemorrhage James Blundell: took inspiration from Leacock in performing transfusions for women suffering from postpartum hemorrhages Late 1800s Emil Ponfick: RBC lysis in a blood of a woman who died after sheep blood transfusion Leonard Landois: Human RBCs lyse when mixed in vitro with sera of other animals 1869 Braxton Hicks: NaPO4 as an anticoagulant 20th Century 1901: Karl Landsteiner ABO Blood Group System 1902: Anthony Decastello and Adriano Sturli AB blood group 20th Century Edward E. Lindemann: vein-to-vein transfusion Unger: syringe-valve apparatus, more practical 1907 Richard Weil: 1st to perform ABO typing and began compatibility testing, 1st to suggest ABO inheritance Refrigeration of citrated blood prior to use

1913 Reuben Ottenberg: stressed importance of compatibility testing 1914 Albert Hustin: sodium citrate as an anticoagulant solution 1915 Richard Lewisohn: minimum amount of citrate needed for anticoagulation 1916 Rous and Turner: introduction of citrate dextrose solution for RBC preservation 1924 Felix Bernstein: proof of inheritance of blood groups Issues in race distribution 1927 American Association of Immunologists: adopted the current ABO terminology proposed by Landsteiner 1939-1940 Philip Levine (together with Stetson, Landsteiner and Alex Wiener): 1st discovery of Rh blood groups 1941 Charles Drew: developing techniques in blood transfusion and blood preservation during WWII 1943 Loutit and Mollison: acid-citrate dextrose formula 1945 Robin Coombs, Rob Race and Arthur Mourant: (re)discovery of anti-human globulin (AHG) sera and antiglobulin test which was first described by Carlo Moreschi in 1908 1947 Rh immune Globulin for prevention of Hemolytic Disease of the Fetus and Newborn 1951 Edwin Cohn: development of cell separator, paved the way for component therapy Carl Walter: blood collection using a collapsible bag of polyvinyl resin 1957 Gibson: introduction of citrate-phosphate-dextrose (CPD) 1965 Judith Pool: concentrated factor VIII found in the cryoprecipitate portion of plasma 1968 Brinkhous and Shanbrom: pooling of plasma units to produce factor VIII concentrationAdverse effects of Transfusion 1970s Transition to an all-volunteer blood supply Availability of commercial testing for HBV 1980s The first serologic test to detect HIV was implemented by blood banks to protect the blood supply Opened the possibility of transmission of blood-borne pathogens other than HBV, HCV, and HIV through blood transfusionNon-infectious effects of TransfusionTransfused leukocytes were found to have a number of undesirable effects Graw and colleagues GVHD prevention by blood component irradiation Greenwalt and colleagues first generation leukocyte filter in the prevention of febrile transfusion reactionsBlood Banking in the Modern World Discovery of West Nile Virus in 2002 as a new transfusion-transmitted pathogen Regulation of bacterial contamination on blood components Blood substitutes or blood alternatives

Areas of RBC Biology1. Normal chemical composition and structure of RBC membrane 2. Hemoglobin structure and function3. RBC metabolism

RBC MembraneA semipermeable lipid bilayer supported by a protein meshlike cytoskeleton structure 1. Phospholipids arranged in bilayer structure (40%)2. Proteins integral and peripheral (52%)3. Carbohydrates 8%

Deformability Factors affecting RBC deformability 1. Loss of ATP levels = decrease in spectrin phosphorylation 2. Calcium accumulation or increase in deposition

Permeability RBC membrane is permeable to water and anions (Cl- and HCO3-) but impermeable to cations (Na+ and K+) To maintain RBC volume and water homeostasis, intracellular concentrations of Na+ and K+ are controlled by utilizing ATP

RBC Metabolism Mainly ANAEROBIC Glycolysis breakdown of glucose to generate energy for RBCs

1. Glycolytic (Embden-Meyerhof) Pathway2. Pentose Phosphate Pathway3. Methemoglobin Reductase Pathway4. Leubering-Rapaport Pathway

Hemoglobin Structure and FunctionHemoglobin Synthesis1. Adequate iron delivery and supply2. Adequate synthesis of protoporphyrins3. Adequate globin synthesis

Types in normal adults1. HbA (two alpha, two beta chains; 92-95%)2. HbA2 (two alpha, two delta chains; 2-3%)3. HbF (two alpha, two gamma chains; 1-2%)

Three functions1. Transport of O2 from the lungs to tissues2. Transport of CO2 from tissues to the lungs 3. Buffering of blood

Hemoglobin function Oxygen delivery to tissues 2-3 DPG Tense form lower High affinity to oxygen Relaxed form higher High affinity to oxygen

Hemoglobin-oxygen dissociation curve Importance: It permits a considerable amount of oxygen to be delivered to the tissues with a small drop in oxygen tension

Ligands1. H+ ions2. CO23. Organic phosphates (2,3 DPG) Shift to the right Increased 2,3 DPG = decrease High affinity to oxygen = increase oxygen delivery to tissues

RBC PreservationRBC viability Measure of in-vivo RBC survival following transfusion 75% of cells transfused should remain viable for 24 hours Liquid state at 1-6 degrees Celsius for a specific number of days

Storage lesion Loss of RBC viability associated with various biochemical changes Decrease in pH, decrease in glucose consumption, decrease ATP levels, buildup of lactic acid, loss of RBC function

Anticoagulant Preservative SolutionsName Storage timeAcid-citrate-dextrose (ACD) 21 daysCitrate-phosphate-dextrose (CPD) 21 daysCitrate-phosphate-double dextrose 21 daysCitrate-phosphate-dextrose-adenine (CPDA-I)35 days

Additive Solutions Preserving solutions that are added to the RBCs after removal of plasma with/without platelets Beutler development Lovric and Hogman implementationa. Lovric CP2D and additive solution (saline, adenine, glucose, trisodium citrate, citric acid, sodium phosphate)b. Hogman CPD and additive solution (saline, adenine, glucose (SAG), and later with mannitol (SAGM)

RBC freezing For autologous units and storage of rare blood types -65 C, 10 years Glycerol

RBC Rejuvenation Restoration of ATP and 2,3 DPG levels PIGPA PIPA Rejuvesol used to salvage liquid-stored RBCs that have reached outdate