What is Immunohematology (Blood
Banking)?
 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: sheep’s 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
 20 Century
th
- 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
concentration
Adverse 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
transfusion
Non-infectious effects of Transfusion
Transfused 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
reactions
Blood 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 Biology
1. Normal chemical composition and structure
of RBC membrane
2. Hemoglobin structure and function
3. RBC metabolism
RBC Membrane
A 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) Pathway
2. Pentose Phosphate Pathway
3. Methemoglobin Reductase Pathway
 4. Leubering-Rapaport Pathway
Hemoglobin Structure and Function
Hemoglobin Synthesis
1. Adequate iron delivery and supply
2. Adequate synthesis of protoporphyrins
3. Adequate globin synthesis
Types in normal adults
1. 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 functions
1. Transport of O2 from the lungs to tissues
2. 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
 Ligands
1. H+ ions
2. CO2
3. Organic phosphates (2,3 DPG)
 “Shift to the right”
- Increased 2,3 DPG = decrease High
affinity to oxygen = increase oxygen
delivery to tissues
RBC Preservation
RBC 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 Solutions
Name Storage time
Acid-citrate-dextrose (ACD) 21 days
Citrate-phosphate-dextrose (CPD) 21 days
Citrate-phosphate-double dextrose 21 days
Citrate-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 implementation
a. 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