ABO Blood Group System

ABO Blood
Group System
THE ABO BLOOD GROUP SYSTEM

❖ Most important of all blood groups in transfusion practice

❖ It is the only blood group system in which individuals have antibodies in their
serum to antigens that are absent from their RBCs. This occurs without any
exposure to RBCs through transfusion or pregnancy (naturally occurring)
➢ EX. BLOOD GROUP/TYPE A → ANTI-B

❖ Due to the presence of these antibodies, transfusion of an incompatible ABO type

may result in immediate lysis of donor RBCs. This produces a very severe, if not
fatal, transfusion reaction in the patient
❖ Testing to detect ABO incompatibility between a donor and potential transfusion
recipient is the foundation on which all other pre-transfusion testing is based.
 1901 – ABO discovered
HISTORICAL PERSPECTIVE

 Karl Landsteiner → Discovery of the first

human blood group system.
 First to conduct reverse and Forward
typing
 A,B, and O Blood group
 Sturli and Von Decastello → AB Blood group
 I gene – code for the ABO allele’s:
1. IA
2. IB
3. i
THE LANDSTEINER LAW

❖ The antigen is present on the RBC surface and determines the blood group/type
❖ The corresponding antibody is NEVER FOUND in the individual’s serum
❖ The OPPOSITE naturally occurring antibody is always present in the individual’s serum

Blood Antigens on the Naturally Percentage in the

groups RBC surface occurring antibody American
in serum population
A A Anti-B 40%
B B Anti-A 10%
AB A and B None 5%
O H Anti-A and Anti-B 45%
 Importance of ABO

✓ Almost all normal healthy individuals above 3-6 months of age have
“naturally occurring Abs” to the ABO antigens that they lack

✓ These Abs termed naturally occurring because they were thought to

arise without antigenic stimulation
 Importance of ABO

✓ These “naturally occurring” Abs are mostly IgM class.

✓ Thatmeans that, they are Abs capable of agglutinating saline/ low

protein suspended red cell without enhancement and may
activate complement cascade; can cause strong rapid
intravascular hemolysis
 Inheritance of the ABO blood group

Theoryfor Inheritance of the ABO blood group was first described

by BERNSTEIN in 1924

He demonstrated that an individual inherits one ABO gene from

each parent (CODOMINANCE) and these two genes determine
which ABO antigens are present on the RBC membrane

One position, or locus, on each chromosome 9 is occupied by an A,

B, or O gene
 Inheritance of the ABO blood group

The O gene is considered an amorph, as no detectable antigen is

produced in response to the inheritance of the genes

AA, BO and OO – denotes genotype

A and B – denotes phenotype
Inthe case of Blood type O individual they possess homozygous OO
genotype. That’s why the phenotype is Blood type O
Inheritance of ABO antigens from PARENTS

Parent A B O
alleles
A AA AB AO

B AB BB BO

O AO BO OO
Parents’ blood type is O and A, what are the possible blood types
of their offspring?
Parents’ blood type is B and AB, what blood type would not be
expected in their offspring?
 Formation of A, B and H Red cell antigen

Formation of ABH antigens results from the interaction of genes of

three separate loci (ABO, Hh and Se)
These genes do not actually code for the production of antigens but
rather produce specific enzymes (glycosyltransferase) that add
sugars to a basic precursor/precursor substance called
paragloboside or glycan

GENE → ENZYME → SUGAR (PS) → ANTIGEN

 Formation of A, B and H Red cell antigen

o H and Se genes - located on chromosome 19

➢ Not part of ABO blood group system; but their inheritance affect the ABO blood
group
➢ H gene
✓ must be inherited to form ABH antigens on the RBCs ;codes for the H antigen
✓ Enzyme produced acts on Type II precursor substance
✓ Most individuals are homozygous HH
✓ The hh genotype is extremely rare and is referred to as the Bombay phenotype
➢ Se gene
✓ must be inherited to form ABH soluble antigens in secretions ; codes for the
H substance
✓ Enzyme produced acts on Type I precursor substance

o ABO genes – located on chromosome 9

 Formation of A, B and H Red cell antigen

o H antigen
➢precursor structure on which A and B antigen are
made
 Formation of A, B and H Red cell antigen

The precursor substance on erythrocytes is referred as Type

2 precursor substance. Means that the terminal galactose
on the precursor substance is attached to the N-
acetylglucosamine in a beta 1-4 linkage

Type 1 precursor substance means that the terminal

galactose on the precursor substance is attached to the N-
acetylglucosamine in a beta 1-3 linkage
 Formation of A, B and H Red cell antigen

❖ Type I and Type II precursor substances; both are composed of 4 sugars:

➢ 2 molecules of D-galactose
➢ 1 molecule of glucose
➢ 1 molecule of N-acetylglucosamine

❖ Differ only in the linkage of the terminal sugars (terminal D-galactose is attached to N-
acetylglucosamine)

❖Type 1 precursor substance is water soluble and is found in

plasma and in tissues and body secretions ; glycoproteins
❖Type 2 precursor substance is fat soluble and is found as an
integral part of the red cell membrane ; glycolipids
Type 2 precursor substance
H Antigen

α-2-L-
fucosyltransferase

L-fucose L-fucose, will

attached to
TYPE 2 PS
 A Antigen

A gene codes for the production of α-3-

N-Acetylgalactosaminyltransferase

Will attached to the

H antigen to
expressed A
N-Acetylgalactosamine
antigen
B Antigen

B genes code for the D-galactose will

production of α-3-D- attached to H antigen to
galactosyltransferase expressed B Antigen
 OCCUPIED H ANTIGENIC SITES BLOOD TYPE

810,000 to 1,170,000 A

610,000 to 830,000 B

A: 600,000 AB
B: 720,000

None O
NOTES

✓Amount of H-antigen / reaction with anti-H:

✓O > A2 > B > A2B > A1 > A1B
 ABO ANTIBODIES

✓ ABO antibodies is initiated at birth, but titers are generally too low

✓ ABO testing before 3-6 months of age cannot be considered valid

because some or all antibodies present maybe IgG maternal
antibodies

✓ More
logical to perform forward typing on cord blood from
newborns
 ABO ANTIBODIES

✓ Antibody production peak: 5-10 years of age

✓ Decline: later in life

✓ Elderlypeople usually have lower levels of Anti-A and Anti-B;

therefore antibodies may be undetectable in Reverse grouping.
 ABO ANTIBODIES

➢They 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 (H specificity) and other
ABO blood groups depending on the amount of H antigen (anti-H
lectin)
REMEMBER!

GROUP Antigen Antibody

AB
 NOTES:

✓ L-fucose is the sugar responsible for H specificity (blood group O)

✓ Amount of H-antigen/reaction with anti-H: O > A2 > B > A2B > A1 > A1B
✓ The term Bombay has been used to refer to the phenotype that lacks normal expression
of the ABH antigens because of the inheritance of the hh genotype.
✓ The hh genotype does not elicit the production of α-2-L-fucosyltransferase. Therefore, L-
fucose is not added to the type 2 chain, and H antigen is not expressed on the RBC

BLOOD GROUP “O” BOMBAY PHENOTYPE (hh)

REACTION WITH ANTI-H
4+ 0 (NEGATIVE)
4+ ONE SOLID AGGLUTINATE

3+ SEVERAL LARGE AGGLUTINATE;

CLEAR BACKGROUND

2+ MEDIUM SIZED AGGLUTINATES;

CLEAR BACKGROUND

1+ SMALL AGGLUTINATES TURBID

BACKGROUND

W+ TINY AGGLUTINATES TURBID

BACKGROUND

0: NO AGGLUTINATION OR NO
HEMOLYSIS
 Two Methods in determining blood type

oForward/Direct/Cell blood typing

✓Defined as using known sources of commercial anti-sera (anti-A and anti-B) to detect
antigens on an individual RBC
➢ Specimen: Patient’s red cell (RCS)
➢ Reagent: Anti-sera
oReverse/Indirect/Back blood typing
✓Defined as detecting ABO antibodies in the patient’s serum by using known reagent
RBC’s namely A1 cells and B cells (Known cells)
✓Only unique to the ABO blood group system; checks results of forward typing
➢ Specimen: Patient’s serum/plasma
➢ Reagent: Known cells
Procedure in Blood Typing

FORWARD TYPING REVERSE TYPING

Reagents for Forward typing
Anti A
Anti B
Anti A,B
Reagents for reverse typing
A1 cells
B cells
O cells
FORWARD/DIRECT TYPING
REVERSE/INDIRECT TYPING
TEST YOURSELF!

FORWARD REVERSE
Anti A Anti B A1 cells B cells
Patient
Forward:
Reverse:
 SERUM Group O

Type O individuals contains Anti-A, Anti-B and as wells Anti-A,B

Anti-A,B – will react with A and B cells. Originally thought to be just a

mixture of anti-A and anti-B

Anti-A,B is not part of ABO testing, but some believe that anti-A,B is
more effective at detecting weakly expressed A and B antigens than
Anti-A and Anti-B.
INTERACTIONS OF THE Sese, Zz and ABH GENES

 Sese system
➢ regulatesthe formation of H antigen and subsequently, of A and B
antigens in secretory cells (ABH soluble substances)
➢ Genotypes: SeSe, Sese, sese
✓Secretors: SeSe , Sese
✓Non-secretors: sese

 Zz system
➢ regulates production of H antigens on erythrocytes
DISTINCTION OF ABH ANTIGENS AND ABH SOLUBLE SUBSTANCES

ABH Antigens ABH Soluble substances

RBC, epithelial cells, All body secretions except

Found where? platelets, CSF
lymphocytes,
endothelial cells,

Secreted substances Glycolipids Glycoprotein

1st sugar in the precursor Glucose N-acetylgalactosamine
substance
Precursor chain Type 2 Type 1
Linkage Beta 1-4 linkage C1-C3 (Beta 1-3 linkage)
Regulating Gene Zz gene Se gene
Test for ABH soluble substances/Determination of secretor status

SALIVA NEUTRALIZATION TEST

Principle: Hemagglutination Inhibition
(+) reaction: without agglutination
(-) reaction: with agglutination
Test for ABH soluble substances/Determination of secretor status

SALIVA NEUTRALIZATION TEST

Principle: Hemagglutination Inhibition
(+) reaction: without agglutination = soluble substance
is present
(-) reaction: with agglutination = soluble substance is
absent
Protocol:

1. Saliva heated to boil (560C for 10 mins to denature salivary

amylase to prevent interference ; prolonged heating may
cause destruction of soluble substance)
2. Dilute saliva using NSS (1:2) to reduce non-ABH soluble
substances present in saliva
3. Dilute anti-sera (1:4) to reduce potency not to cause false
positive reaction
  B substance is present because it has
2 steps procedure been neutralized first by anti sera,
not reacting to the B cell = without
agglutination
 A substance is not present because
anti A was not neutralized, so anti A
Step 1: Saliva + Antisera/ Lectin reacts with a cell – agglutination
Step 2: Mixture of step 1 + Known Red cell suspension

Example result:  Therefore, The saliva is from an

individual which is a B secretor
Antisera A cell B cell O cell
with
Saliva
Anti – A + - -
Anti – B - - -
Anti - H - - -
 2 steps procedure

Step 1: Antisera/ Lectin only

Example result:

Antisera A cell B cell O cell

with
Saliva
Anti – A + - -
Anti – B - - -
Anti - H - - -
 2 steps procedure

Step 2: Mixture of step 1 + Known Red cell suspension

Example result:

Antisera A cell B cell O cell

with
Saliva
Anti – A + - -
Anti – B - - -
Anti - H - - -
END