HEM 2133

Immunohaematology I
Lesson 4: Isoantibodies of ABO
System

Blood Group Antigens

Made of glycoprotein and glycolipid molecules
on surface of RBC
Normal part of RBC plasma membrane
E.g. Person with blood group A will have
antigen A on RBC plasma membrane
Genetically determined (i.e. inherited from
mother and father)

Antigens of ABO system

Each individual inherits one ABO gene from
each parent
These two genes determine which Ags are
present on RBC membrane
One position or locus on each chromosome
number nine is occupied by an A, B or an O
gene

 A locus termed H and the final product of the

genes at that locus, H antigen, is necessary for
the expression of normal ABO antigens

 The ABO genes do not code for the production

of ABO antigens, but rather produce specific
glycosyl transferases
Glycosyl transferases add sugars to a basic
precursor substance on the RBCs
The inheritance of at least one H gene (HH or
Hh) obtain the production of an enzyme called
-2-L-Fucosyl transferase, which transfers the
sugar to the terminal galactose of the
precursor chain

 The H substance must be formed for the other

sugars to be attached in response to an
inherited A and/or B genes

ABO Genetics
Genes at three separate loci control the
occurrence and location of A and B antigens
1. Hh genes H and h alleles
H allele codes for a fucosyltransferase
enzyme that adds a fucose on precursor
substance to form the H antigen onto which
A and B antigens are built on red blood cells
h allele is a silent allele (amorph)

2. Se genes Se and se alleles

Se allele codes for a fucosyltransferase
enzyme that adds fucose to precursor
substance in secretory glands
Controls expression of H antigens in secretions
(i.e. saliva, body fluids, etc)
se allele is an amorph
Secretor gene controls only the presence or
absence of the H substance in body secretions
It does not affect the presence of the H
substance on erythrocytes

3. ABO genes A, B and O alleles

A and B alleles code for glycosyltransferase (a
fucosyltransferase enzyme that add a sugar
onto H antigens to produce A and B antigens)
O allele does not code a functional enzyme

Hh gene H and h alleles Controls presence of H, A

(h is an amorph)
and B antigens on both
RBCs and in secretions

Se gene Se and se
alleles (se is an amorph)

Controls presence of H
antigen in the secretions

ABO genes A, B and O

alleles

Inherit 1 gene from each

parent that codes for an
enzyme that adds a sugar
to the H antigen

H Antigen
The H gene codes for an enzyme
(fucosyltransferase) that adds a fucose to the
terminal sugar of a precursor substance
H antigen is the foundation upon which A and
B antigens are built
A and B genes code for enzymes that add an
immunodominant sugar to the H antigen

Formation of the A Antigen

The A gene codes for an enzyme that adds
GalNAc (N-Acetyl-D-galactosamine) to the
terminal sugar of the H Antigen

Formation of the B Antigen

B gene codes for an enzyme that adds DGalactose to the terminal sugar of the H
antigen

 The H antigen is found on the RBCs when

there is an Hh or HH genotypes but not with
the hh genotype
The A antigen is found on the RBCs when
there is Hh, HH and A/A, A/O or A/B
genotypes
The B antigen is found on the RBCs when
there is Hh, HH and B/B, B/O or A/B genotypes

Amount of H Antigen According to

Blood Group
Blood group O people have red blood cells rich
in H antigen
Neither the A or B genes have converted the H
antigens to A or B antigens just a whole
bunch of H
Amount of H antigen
O > A2 > B > A2B > A1 >A1B

Subgroups of ABO
Subgroups differ in the amount of antigen
expressed on the red cell membrane (e.g. A1
red cells express about 5 times more A antigen
than A2 red cells)
Some subgroups highly branched, complex
antigenic structures
Some subgroups simplified linear antigenic
structures
Most common subgroups A

Subgroups of A
A1 and A2
20% of group A and AB person belong to
subgroup A2 and 80% to A1 (both types of red
cells react with anti-A)
A failure to classify a weak subgroup of A may
lead the donor to be classified as a group O
and to be transfused to a group O patient
All group O donors must be tested with anti-A,
B to confirm that they are not actually weak
subgroups of A

Other ABO Subgroups

The other weaker subgroups of A exist
(extremely rare) : A3, Aint, Am, Ax, Ael
Subgroups of B even rarer

Bombay Phenotype
Normal people inherit a pair of H genes at a
locus separate from the ABO locus
The classic Bombay phenotype would be
expected to arise most frequently in the
children resulting from consanguineous
marriages
Results in inheritance of an h gene from both
parents at the H locus

 Unable to produce the H glycosyltransferase

Unable to produce H antigen
Regardless of the A, B or O genes inherited by
the individual , no A, B or H antigen is formed
In a homozygous hh individual, even if ABO
genes are present, the precursor for the
synthesis of these antigens is not produced in
the absence of H gene

 This rare blood group is called Bombay group

(Oh)
Typed as group O persons with anti-A and
anti-B sera
Only anti-H can detect the Bombay blood
group
(other groups will give a positive reaction with
anti-H serum, the Bombay group will give a
negative reaction)

Antibodies of ABO system

Anti-A

Arises in sera of group B people

Will agglutinate the RBCs of all group A and
AB people

Most of the anti-A is IgM, although small
amount of IgG and IgA may be present

Anti-A can functionally split into anti-A1, which
react with A1cells but not with A2cells

 A1 antigen is more branched and therefore

reacts differently with anti-A than the lessbranched A2 antigen
A reagent made from the plant Dolichos
biflorus (lectin) can easily differentiate
between A1 and A2 cells

 Anti-B

Serum from group A people contains an
antibody that agglutinates essentially all
group B and group AB RBCs

This antibody also readily agglutinates cells
suspended in saline, activates complement
and may rapidly destroy incompatible RBCs by
intravascular hemolysis

 Anti-A, B

Found in sera of all group O people

Used to confirm group O donors and aid in
identification of weak subgroups of A and B
Anti-H

May be found as a weak, cold-reacting antibody
in the sera of group A1 and A1B people

Found in the sera of people expressing the
Bombay phenotype

When tested with anti-H or U. europaeus lectin,
the RBCs of the Bombay phenotype are negative

Universal Donor
Group O
The red blood cells lack both A and B antigens
Group O donor red cells can be used in times
of urgency or emergency release of donor
units

Universal Recipient
Group AB
Lacks circulating ABO antibodies in plasma
May receive transfusion of RBCs from any ABO
phenotype
Beware: other antibodies may be present