HISTORICAL PERSPECTIVES
papers to blood preservation.
 In 1492, 1st time a blood transfusion was recorded in
history. The blood was taken from three young men and
given to the stricken
 Pope Innocent VII Cosimo de' Migliorati, in the hope of
curing him, unfortunately, all four died.
 Clotting was the principal obstacle to overcome
 Attempts to find a nontoxic anticoagulant began in 1869,
when Braxton Hicks recommended sodium phosphate.
 This was perhaps the first example of blood preservation
research.
 Karl Landsteiner in 1901 discovered the ABO blood
groups and explained the serious reactions that occur in
humans as a result of incompatible transfusions.
 Next came devices designed for performing the
transfusions. Edward E. Lindemann was the first to
succeed.
 He carried out vein-to-vein transfusion of blood by using
multiple syringes and a special cannula for puncturing the
vein through the skin.
 It was not until Unger designed his syringe-valve
apparatus that transfusions from donor to patient by an
unassisted physician became practical.
 An unprecedented accomplishment in blood transfusion
was achieved in 1914, when Hustin reported the use of
sodium citrate as an anticoagulant solution for
transfusions.
 Later, in 1915, Lewisohn determined the minimum
amount of citrate needed for anticoagulation and
demonstrated its nontoxicity in small amounts.
 Transfusions became more practical and safer for the
patient. The development of preservative solutions to
enhance the metabolism of the RBC followed.
 Glucose was tried as early as 1916, when Rous and
Turner introduced a citrate-dextrose solution for the
preservation of blood.
 However, the function of glucose in RBC metabolism was
not understood until the 1930s. Therefore, the common
practice of using glucose in the preservative solution was
delayed.
 World War II stimulated blood preservation research
because the demand for blood and plasma increased.
 The pioneer work of Dr. Charles Drew during World
War II on developing techniques in blood transfusion and
blood preservation led to the establishment of a
widespread system of blood banks.
 In February 1941, Dr. Drew was appointed Director of
the 1st American Red Cross Blood Bank at Presbyterian
Hospital. The pilot
 The pilot program Dr. Drew established became the
model for the national volunteer blood donor program of
the American Red Cross.
 In 1943, Loutit and Mollison of England introduced the
formula for the preservative acid-citrate-dextrose
(ACD).
 Efforts in several countries resulted in the landmark
publication of the July 1947 issue of the Journal of
Clinical Investigation, which devoted nearly a dozen
 Hospitals responded immediately, and in 1947, blood
banks were established in many major cities of the United
States; subsequently, transfusion became commonplace.
 The daily occurrence of transfusions led to the discovery
of numerous blood group systems.
 Antibody identification surged to the forefront as
sophisticated techniques were developed.
 In 1957, Gibson introduced an improved preservative
solution called citrate phosphate-dextrose (CPD), which
was less acidic and eventually replaced ACD as the
standard preservative used for blood storage.
 Frequent transfusions and the massive use of blood soon
resulted in new problems, such as circulatory overload.
Component therapy has solved these problems.
 Before, a single unit of whole blood could serve only one
patient. With component therapy, however, one unit may
be used for multiple transfusions.
 Today, physicians can select the specific component for
their patient’s particular needs without risking the
inherent hazards of whole blood transfusions.
 Physicians can transfuse only the required fraction in the
concentrated form, without overloading the circulation.
 Appropriate blood component therapy now provides more
effective treatment and more complete use of blood
products.
 Extensive use of blood during this period, coupled with
component separation, led to increased comprehension of
erythrocyte metabolism and a new awareness of the
problems associated with RBC storage.
PERMEABILITY
 The permeability properties of the RBC membrane and
the active RBC cation transport prevent colloid hemolysis
and control the volume of the RBC.
 Any abnormality that increases permeability or alters
cationic transport may decrease RBC survival.
 The RBC membrane is freely permeable to water and
anions. Chloride (Cl–) and bicarbonate (HCO3–) can
traverse the membrane in less than a second.
 Experiments with blood transfusion, the transfer of blood
or blood components into a person’s blood stream, have
been carried out for hundreds of years.
 Many patients have died and it was not until 1901, when
the Austrian Karl Landsteiner discovered human blood
groups, that blood transfusions became safer.
FATAL
 Mixing blood from two individuals can lead to blood
clumping or agglutination.
 The clumped red cells can crack and cause toxic
reactions. This can have fatal consequences.
 Karl Landsteiner discovered that blood clumping was an
immunological reaction which occurs when the receiver
of a blood transfusion has antibodies against the donor
blood cells.
 Landsteiner’s work made it possible to determine blood  Antigen - antibody reactions important to
types and thus paved the way for blood transfusions to be immunohematology it involve the agglutination of
carried out safely. erythrocytes by antibodies.

 For this discovery he was awarded the NOBEL PRIZE  Hemagglutination is agglutination of the RBCs. It occurs
in Physiology or Medicine in 1930. in two stages.

 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.  Sensitization occurs when the antigen-binding sites of the
1. The RBC contain hemoglobin - a protein that binds antibodies become closely associated with the antigenic
oxygen. RBC’s transport oxygen to, and remove carbon determinants of the RBC membrane. The antibodies and
dioxide from, the body tissues. binding sites are held together loosely by non-covalent
2. The WBC fight infection. bonds.
3. The PLATELETS help the blood to clot, if you get a  Visible agglutination occurs when several RBC’s are
wound for example. physically joined together by the antigen-antibody union.
4. The PLASMA contains salts and various kinds of This stage depends on many factors, such as the pH,
proteins. temperature, and ionic strength of the suspension
 Study of transmission of inherited characteristics. medium.

 Important in the study of antigen inheritance and

inherited disorders. 1. O O O O
DNA
A AO AO A AO AO
 Double Helix
O OO OO A AO AO
 BASES: adenine (A), thymine (T), cytosine (C), and
guanine (G).
2. B B B O
 Gene is a segment of DNA arranged along the
chromosome at a specific position called locus. A AB AB A AB AO

B BB BB B BB BO

 Gene at specific locus that differ in their nucleotide

1. The use of proteolytic enzymes such as papain or ficin
sequence are called alleles.
can enhance cell-to-cell contact.
 Homozygous (Identical alleles) 2. RBC’s sensitized by incomplete antibodies (antibodies
that will not react in saline) agglutinate when antiserum
 Heterozygous (Nonidentical alleles) against human IgG is added.

 The amount of available antigen and antibody also affects

hemagglutination.
 DOSAGE EFFECT: presence of homozygous genotype
can express itself with more antigen than the  PROZONE - occurs when antibody molecules are in
heterozygous genotype. excess of available antigenic sites, resulting in false-
negative reactions.
 GENOTYPE: Total genetic composition of an
individual, representing maternally and paternally derived  EQUIVALENCE - optimal proportions of antigen and
genes. antibody present-allows hemagglutination to occur.
 PHENOTYPE: Detectable or expressed characteristics of  Antigen excess (post-zone) also results in false-negative
genes. reactions.
 RBC have antigens; Plasma/ Serum have antibodies. LOCUS
ANTIGENS CHROMOSOME

 “Antibody generator” - Proteins, glycoproteins or 1 Rh, Duffy

glycolipids found in the serum, plasma, RBC’s or other
cells that can induce antibody production.
4 MNS
ANTIBODY
7 Kell
 Gamma globulins that act against the antigen or bind the
antigen. They may be naturally occurring or may be
9 ABO
immunogenic.

 Antibodies to the blood group antigens maybe of IgM, 18 Kidd

IgG or IgA in nature.

 May be warm reactive or cold reactive antibodies. 19 H, LEWIS, LUTHERAN

22 P1
ABO BLOOD GROUP SYSTEM  On the 37th day of fetal life, attachment of
immunodominant sugars occurs on the RBC membrane
 Most important system in transfusion and transplantation and it is dependent on ABH genes inherited.
therapy - the only blood group that with naturally
occurring antibodies in serum against antigens that are ANTIGE STRUCTURE
absent from their RBC’s. N
 ANTIGEN INHERITANCE H
- ABH genes are located on chromosome 9

- ABH genes are inherited in a codominant manner

following simple Mendelian Genetics Laws
B
EXAMPLE

1. Parent blood type - O x A

 Possible blood type - O and A only of children
exclusion AB and B
A
2. Parent blood type - B x AB
 Possible blood type - A, B, AB of children
 Exclusion - O

INTERACTIONS OF THE Sese, Zz and

PARENT A B O
ALLELES

THE FORMATION OF A, B, AND H A AA AB AO

(A) (AB) (A)
ANTIGENS
B AB BB BO
 ABO genes code not for the antigen themselves but for (AB) (B) (B)
the production of glycosyltransferases that add
immunodominant sugars to a basic precursor substance. O AO BO OO
(A) (B) (O)
GENE GLYCOSYLTRANSFERA IMMUNODOMINA
BLOODTYPE/
SE
GENOTYPE
NT SUGAR ABH GENES
PHENOTYPE
H gene L-fucosyltransferase L-fucose Sese system - regulates the formation of H antigen and
A gene O N-acetylgalactosaminyl- OO N-acetyl-D- subsequently, of A and B antigens in secretory cells
transferase galactosamine
B gene A D-galactosyltransferase AA, AO D-galactose Zz system - regulates production of H antigens on
B BB, BO erythrocytes

AB AB T DISTINCTION OF ABH ANTIGENS
he presence of H substances in body secretions is AND ABH SOLUBLE SUBSTANCES
controlled by the Se gene. Individuals who are
ABH ABH SOLUBLE
homozygous (Se Se) or heterozygous (Se se) for this gene
ANTIGENS SUBSTANCES
are called secretors (approximately 80% of the
population).
Found where? In red cells, In all body secretions
 Group O secretors have H antigen in their secretions.
epithelium
 Group A secretors have A and H antigens in their
tissues, BM,
secretions.
other cells
 Group B secretors have B and H antigens in their
secretions. Secreted Glycolipids Glycoproteins
 Group AB secretors have A, B and H antigens in their substances
secretions.
1st sugar in the Glucose N - acetylgalactosamine
 DEVELOPMENT – A and B antigens begin to develop precursor
in the sixth week of fetal life, but do not reach adult substance
levels until 3 years of age. Antigen levels approximate
50% at birth. Precursor chain Type 2 Type 1 AND Type 2

Linkage 1à4 linkage 1à3 linkage

Regulating FUT1 (Zz gene) FUT2(Se gene)

Gene

 Because the O gene does not result in the conversion of H

ABO ANTIBODIES
antigen to other antigens, the RBC’s of group O - are mostly naturally occurring antibodies that are
individuals have the highest concentration of H detectable 3 to 6 months after birth following
antigen. exposure to ABO-like antigens in the environment.
 Group AB individuals have the lowest concentration. - are mostly IgM in nature, but some are IgG.
- react best at RT or colder, and activate complement.
 Immune ABO antibodies develop in response to exposure
to ABO-incompatible RBC’s or other sources of exposure
to ABO antigens.

 The immune ABO antibodies are usually IgG and can

cross the placental barrier.

INCIDENCE
GROUP WHITES BLACKS

O 45% 49%

A 40% 27%

B 11% 20%

AB 4% 4%

FORWARD AND REVERSE

GROUPINGS

TESTING
 FORWARD GROUPING - analyzes patient cells for the
presence of ABO antigens. Testing is performed at room
temperature by adding anti-A and anti-B reagents to the
patient’s RBC’s.

 Cell suspensions used in blood banking procedures are

usually 2% to 5%.

 All Group O donors should also be tested with anti-AB

reagent.

 Agglutination is a positive reaction.

 REVERSE GROUPING - analyzes patient serum or

plasma for the presence of anti-A or anti-B antibodies. In
adults, anti-A or anti-B is present in the serum when the
corresponding antigen is not present on the RBC’s.
 Because neonates have not developed these types of
antibodies, they are not candidates for reverse ABO
grouping.
 Testing is performed at room temperature by adding
the patient’s serum or plasma to suspension of known
A1 and B RBC’s.
 Agglutination is the positive reaction
ABO SUBGROUPS
A SUBGROUPS
A2 CELL - (has more antigenic sites for the H antigen thus
giving (+) reaction w/ Anti-H lectin)
A1 CELL - (H antigen sites occupied both A and A1
antigens)
RELATION OF H-SUBSTANCE
(ANTIGEN) & ABO GROUPS
 B SUBGROUPS -are infrequent Anti-B lectin
Bandeiraea siimplicifolia
 BOMBAY (Oh) PHENOTYPE – The allele h is very
rare and does not produce the L-fucose necessary for
the formation of H structure.
 The genotype hh or Hnull is also known as the
Bombay phenotype or Oh.
GENERAL CHARACTERISTICS OF
OH PHENOTYPES
1. Absence of H, A, and B antigens.
2. Presence of Anti-H, Anti-A, and Anti-B in serum.
3. Strong reactivity with Anti-I reagents.
4. A recessive mode of inheritance.
5. No reaction with Anti-H lectin (Ulex europaeus).
TECHNICAL ERRORS RESULTING
TO ABO DISCREPANCIES
1. Inadequate identification of blood samples, test tubes
or slides.
2. Cell suspensions either too heavy or too light.
3. Clerical errors
4. Mix-up of samples
5. Failure to add reagents/failure to follow
manufacturer’s instructions.
6. Contaminated reagents
7. Uncalibrated centrifuge
Type A types - should basically stick to fruits and vegetables
(high carbs / low fat). They have thicker blood than other
blood types, a sensitive immune system, and should not
consume dairy products, animal fats and meats.  They are at a
heightened risk for cardiovascular disease, diabetes and
cancer.
Type B types - should consume a balanced diet (fruits and
vegetables, grains, fish, dairy, meat, but avoid chicken).  They
have the best chance of bypassing or overcoming everyday
types of diseases, including heart disease and cancer.
Type AB types -  should consume a mostly vegetarian diet,
and only on rare occasions some fish, meat (no chicken), and
dairy.
Type O types - should basically stick to a high protein diet
(including red meat), low carbs, and enriched with fruits and
vegetables.  They should limit the intake of wheat germ, whole
wheat products, corn, and avoid dairy products and most nuts.
Type O types are commonly affected with hypothyroidism,
high stomach acid
(leading to ulcers), and thinner blood with greater resistance to
blood clotting.
How do different Blood Types compare to various medical
conditions?
 There are some known blood type / disease-risk
associations, such as pernicious anemia, diabetes, or
certain types of cancer being slightly more prevalent
with Type A compared to Type O individuals, while
in contrast to Type A, Type O individuals have a
marginally higher incidence of ulcers / H. Pylori
infections.  
 Types O and B also have greater susceptibility to
infectious diseases such as scarlet fever, cholera,
typhoid, or the bubonic plague, while Type A shows
greater susceptibility to the smallpox virus, and it is
more prone to blood clotting.  Blood-sucking insects
(that carry diseases) prefer Type O blood
PERSONALITY AND BLOOD TYPES
 Type A’s most often described themselves in ways
related to the following characteristics: sensitive to the
needs of others, good listeners, detail oriented,
analytical, creative and inventive.
 Type B’s often described themselves in ways related to
the following characteristics: subjective, easygoing,
creative, original and flexible. In another study, they
tend to be insightful, mystical, idealistic, globally
-oriented, people-oriented and good at imagining. They
also reported that they learned best through listening,
then reflecting on and interpreting what they had
observed.
 Type O’s most often described themselves in ways
related to the following characteristics: responsible,
decisive, organized, objective, rule-conscious, and
practical. They are also more detail and fact oriented,
logical, precise and orderly than other types.

BLOOD FREQUENCY ANTI- A ANTI- B ANTI- A1 ANTI- H

GROUP lectin

( Dolichos
biflorus)

A1 Approx. 80% ++++ - ++ -

A2 <20% +++ - - ++