TOPIC 6: ABO Blood Group System

IMMUNOHEMATOLOGY; Blood Banking

FEBRUARY 16, 2024

THE ABO BLOOD GROUP SYSTEM NOTE:

➔ This is the most important blood group in transfusion practice. Due to the presence of these
antibodies, transfusion of an
➔ It is the only blood group system in which individuals have
incompatible ABO type may result in
antibodies in their serum to antigens that are absent from their
immediate lysis of donor RBCs. This
red blood cells.
produces a very severe, if not fatal,
➔ Almost all normal healthy individuals above 3-6 months of age transfusion reaction in the patient.
have “naturally-occurring antibodies” to the ABO antigens that
they lack. Testing to detect ABO incompatibility
between a donor and potential
➔ These naturally-occurring antibodies are innately present
transfusion recipient is the
without any exposure to foreign red blood cells through
foundation on which all other
transfusion or pregnancy.
pre-transfusion testing is based.
➔ These antibodies were termed “naturally-occurring” because
they were thought to arise without antigenic stimulation.
➔ These “naturally-occurring” antibodies are mostly IgM class. Karl Landsteiner
— He was the first to perform
➔ IgM antibodies are known for their ability to agglutinate (clump FORWARD and REVERSE BLOOD
together) red blood cells that are suspended in saline (a salt TYPING .
solution) or low-protein solutions. This agglutination can occur
without any need for enhancement.
➔ The activation of the complement cascade by IgM antibodies I Gene — is the code for ABO alleles
can lead to the destruction (hemolysis) of red blood cells. This
IA A
process can be strong and rapid, occurring within the
bloodstream (intravascular) and potentially causing serious IB B
consequences such as anemia.
i O

➔ NOTES TO REMEMBER:
◆ Antigens are present in the red blood cell surface.
◆ Antibodies are present in the serum or plasma.

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THE LANDSTEINER LAW
1. The ANTIGEN is present on the red blood cell surface and will determine the blood group/type.
2. The CORRESPONDING (foreign) antibody is NEVER FOUND in the individual’s serum.
3. The OPPOSITE (naturally-occurring) antibody is ALWAYS PRESENT in the individual’s serum.

BLOOD RED BLOOD CELL SERUM AMERICAN

GROUP ANTIGEN ANTIBODY POPULATION

A A antigen Anti-B 40%

B B antigen Anti-A 10%

A antigen
AB None 5%
B antigen

Anti-A
O H antigen 45%
Anti-B

BLOOD TYPE COMPATIBILITY CHART

NOTES TO REMEMBER:
➔ Blood Type O– is the universal donor.
➔ Blood Type AB+ is the universal recipient.
➔ Blood Group O is the most common blood group among populations.
➔ Blood Group A is the second-most common blood group among populations.
➔ Blood Group AB is the least common blood group among populations.
➔ Blood Group O is the hardest blood group to transfuse due to its selective donor compatibility.

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ABO INHERITANCE PATTERNS
➔ Theory for Inheritance of the ABO Blood Group was first described by FELIX BERNSTEIN in 1924
➔ He demonstrated that an individual inherits one ABO gene from each parent (CODOMINANCE) and
these two genes determine which ABO antigens will be present on the RBC membrane.
➔ The O (also called “H”) gene is considered an AMORPH as no detectable antigen is produced in
response to the inheritance of the genes.

COMPLEX SIMPLE
GENOTYPE PHENOTYPE GENOTYPE PHENOTYPE

A1A1 A1 AA A

A1A2 A1 AO A

A1O A1 BB B

A2A2 A2 BO B

A2O A2 AB AB

A1B A1B OO O

A2B A2B hh Bombay

BB B The term BOMBAY has been used to refer to the

phenotype that lacks normal expression of the ABH
BO B antigens because of the inheritance of the hh gene.
The hh genotype does not elicit the production of
OO O α-2-L-fucosyltransferase. Therefore, L-fucose is not
added to the type 2 chain, and the H antigen is not
hh Bombay expressed on the red blood cell.

➔ H and Se genes are not part of the ABO blood group system,
but the way they are inherited can influence the characteristics of the ABO blood group:
● H gene
— The H gene codes for the H-antigen.
— This must be inherited to form ABH antigens on red blood cell surfaces.
— The enzyme that the H gene produces acts on the Type II precursor substance.
— Most individuals are homozygous HH (Blood Type O).
— The hh genotype is extremely rare and is referred to as the Bombay phenotype.
— The Bombay phenotype (hh) is characterized by a lack of H gene on red blood cell surfaces.
● Se gene (secretory)
— The Se gene codes for the H-substance.
— This must be inherited to form ABH antigens in secretions.
— The enzyme that the Se gene produces acts on the Type I precursor substance.

➔ ABO genes are located on CHROMOSOME 9.

➔ H and Se genes are located on CHROMOSOME 19.

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 INTERACTIONS OF THE Sese, Zz, and ABH GENES
Sese System Zz System
This regulates the formation of H antigen and subsequently,
This regulates the production of H antigens
the formation of A and B antigens in secretory cells
on red blood cells.
(called ABH soluble substances).

THREE GENOTYPES

SECRETORS SeSe
Sese

NON-SECRETORS sese

FORMATION OF A, B, and H RED CELL SURFACE ANTIGENS

➔ The formation of ABH antigens results from the interaction of genes of
Type I and Type II
three separate loci (ABO, Hh and Se).
precursor substances
➔ These genes code for the production of enzymes (glycosyltransferase) are composed of four sugars:
that add sugars to a basic precursor substance called the paragloboside
or glycan. ● D-galactose (x2)
● Glucose (x1)
➔ The H-antigen (O-antigen) is the precursor structure on which A and B
● N-acetylglucosamine (x1)
antigens are made.

TYPE I and TYPE II

TYPE 1 PRECURSOR SUBSTANCE differs only in the
★ The terminal galactose on the precursor substance is attached to the linkage of the terminal sugars
N-acetylglucosamine in a beta 1-3 linkage. (terminal D-galactose
★ Type 1 precursor substance is water soluble and is found in plasma is attached to
and in tissues and body secretions; glycoproteins N-acetylglucosamine)

TYPE 2 PRECURSOR SUBSTANCE

The precursor substance on
★ The terminal galactose on the precursor substance is attached to the
red blood cells is referred to as
N-acetylglucosamine in a beta 1-4 linkage.
Type 2 Precursor Substance
★ Type 2 precursor substance is fat soluble and is found as an integral
(Type 2 paragloboside)
part of the red cell membrane; glycolipids

GLYCOSYLTRANSFERASES AND IMMUNODOMINANT SUGARS

RESPONSIBLE FOR A, B, and H ANTIGENS

CODING GLYCOSYLTRANSFERASE IMMUNODOMINANT BLOOD

ATTACHES TO ANTIGEN
GENE (sugar-transferring enzyme) SUGAR TYPE

H α-2-L-fucosyltransferase L-fucose Type II paragloboside H O

A α-3-N-acetylgalactosaminyltransferase N-acetyl-D-galactosamine H antigen A A

B α-3-D-galactosyltransferase D-galactose H antigen B B

α-3-N-acetylgalactosaminyltransferase N-acetyl-D-galactosamine
A+B H antigen A+B AB
α-3-D-galactosyltransferase D-galactose

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OCCUPIED H ANTIGENIC SITES BLOOD TYPE Amount of H-antigen or the reaction with anti-H is as follows
O > A2 > B > A2B > A1 > A1B
810,000 to 1,170,000 A
This sequence indicates the decreasing amount of H-antigen and the
strength of reaction with anti-H antibodies in different blood types:
610,000 to 830,000 B
➔ Blood type O has the highest amount of H-antigen.
It also has the strongest reaction with anti-H antibodies.
A: 600,000 ➔ A2 and B have less H-antigen compared to O.
AB
B: 720,000 ➔ A2B has less H-antigen than A2 and B.
➔ A1 has less H-antigen than A2 or B.
None O ➔ A1B has the least amount of H-antigen .

ABO ANTIBODIES
➔ ABO antibodies are initiated at birth, but the titers are generally too low.
➔ ABO testing before 3-6 months of age cannot be considered valid because some or all antibodies
present may be IgG maternal antibodies (from the mother).
➔ It is more logical to perform forward blood typing on cord blood from newborns.
➔ Antibody production peaks at 5-10 years of age, and declines later in life.
➔ Elderly people usually have lower levels of Anti-A and Anti-B,
therefore antibodies may be undetectable in reverse blood typing.
➔ Antibodies are present in some animals and plants as lectins.
Lectins are plants or seed extracts diluted to agglutinate specific human blood group antigens.
◆ Dolichos biflorus: agglutinates A1 or A1B cells (anti-A1 lectin)

◆ Bandeiraea simplicifolia: agglutinates B cells (anti-B lectin)

◆ Ulex europaeus: agglutinates O cells and other ABO blood groups depending on the amount of H
antigen (anti-H lectin).

VISUAL AID READING DESCRIPTION

4+ One solid agglutinate

Several large agglutinate;

3+
Clear background

Medium-sized agglutinates;
2+
Clear background

Small agglutinates;
1+
Turbid Bakground

Tiny agglutinates;
W+
Turbid background

No agglutination;
0
No hemolysis

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METHODS IN DETERMINING BLOOD TYPE
TWO METHODS OF DETERMINING BLOOD TYPE
Forward Blood Typing Reverse Blood Typing
Direct Typing Cell Typing Indirect Typing Back Blood Typing
➔ This is also known as “Direct Typing” ➔ This is also known as “Indirect Typing”
or “Cell Typing.” or “Back Blood Typing.”
➔ This uses commercial anti-sera (Anti-A & Anti-B) ➔ This uses known reagent red blood cells (A1 cells
to detect antigens on an individual red blood cell. and B cells) in detecting ABO antibodies in the
patient’s serum

FOCUS: Detects Antigens FOCUS: Detects Antibodies

SPECIMEN: Red Blood Cells SPECIMEN: Serum or Plasma
REAGENT: Anti-sera: REAGENT: Known Cells:
– Anti-A antisera – A1 cells
– Anti-B antisera – B cells
– O cells

FORWARD BLOOD TYPING REVERSE BLOOD TYPING

ANTI-A ANTI-B BLOOD TYPE A1 CELLS B CELLS BLOOD TYPE

4+ 0 A 0 4+ A

0 4+ B 4+ 0 B

4+ 4+ AB 0 0 AB

0 0 O 4+ 4+ O

NOTES TO REMEMBER:

Blood Type O individuals contain Anti-A, Anti-B, and Anti-A,B antibodies in their plasma or serum.

Anti-A,B antisera will react with type-A and type-B red blood cells.

Anti-A,B antisera was originally thought to be just a mixture of anti-A and anti-B antisera.

Anti-A,B antisera was not originally a part of the ABO testing, but some believe that anti-A,B is more effective at
detecting weakly expressed A and B antigens than individual Anti-A and Anti-B antisera.

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DISTINCTION OF ABH ANTIGENS AND ABH SOLUBLE SUBSTANCES
ABH Antigens ABH Soluble Substances

Platelets
Lymphocytes
All body secretions
CAN BE FOUND IN Red blood cells
(except for cerebrospinal fluid)
Epithelial cells
Endothelial Cells

SECRETED SUBSTANCES Glycolipids Glycoproteins

FIRST SUGAR IN
Glucose N-acetylgalactosamine
PRECURSOR SUBSTANCE

PRECURSOR CHAIN Type 2 precursor chain Type 1 precursor chain

LINKAGE Beta 1-4 Beta 1-3 (C1-C3)

REGULATING GENE Zz gene Se genes

TEST FOR ABH SOLUBLE SUBSTANCES

SALIVA NEUTRALIZATION TEST
PRINCIPLE: Hemagglutination Inhibition
(+) REACTION: Without agglutination (a soluble substance is present)
(–) REACTION: With agglutination (the soluble substance is absent)

TWO STEPS PROCEDURE:

Step 1: Saliva + Antisera/Lectin
Step 2: Mixture of Step 1 + Known Red Cell Suspension