ABO BLOOD GROUP SYSTEM

LESSON 1: ABO BLOOD GROUP SYSTEM

Introduction:

 2 most clinically significant blood group systems are:

1. ABO Blood Group System
2. Rh Blood Group System

ABO BLOOD GROUP SYSTEM

 Most clinically blood group system in transfusion and transplantation therapy

 It is a unique blood group system because it is the only blood group system in which antibody is
formed by the individual against antigen that is not found on the BRC
 The only system with reverse and indirect blood typing
o Incompatibility with this blood group system  immediate hemolytic transfusion
reaction
 Most severe type of transfusion reaction  immediate death in minutes

GENETIC CONCEPT IN ABO INHERITANCE

 ABO Genes are located in Chromosome 9’s long arm

Bernstein, 1924

 He explained the manner of inheritance of the ABO genes in the long arm of chromosome 9
 According to him, each parent contributes half of the genetic information to the child in the unit
called “gene”
 CODOMINANCE = 50% haplotype comes from the mother and the other 50% comes from the
father.
o Possible blood group of the offspring is readily predicted by punnett square

Landsteiner Law = discovered ABO blood group system

1. The antigen on the RBC determined the blood groups

o Ex: A antigen = A blood type
2. The corresponding antibody is never found in the individual serum
o Ex: A blood type = NO A antibody = B antibody
3. The opposite antibody is always present in the individual’s serum
 o Ex: A blood type = B antibody

Blood Type / Phenotype Genotype

O OO (Homozygous)
A AO, AA ( heterozygous or homozygous )
B BO, BB ( heterozygous or homozygous )
AB AB (two genes)
 O genes
o Amorph = no detectable antigens which is produced in response to the inheritance of
genes

Blood Type O

 Homozygous O
 Autosomal recessive traits
 Inheritance of 2 O genes = non functional

IMPORTANT NOTE:
For antigens that are expressed in:
 Homozygous Manner
o Dosage effect = Double dose = stronger reaction in hemagglutination reactions
 Usually 4+
 Heterozygous Manner
o Single dose

NOMENCLATURE:

Landsteiner Jansky Moss

O I IV
A II III
B III II
AB IV I

ISBT (International Society of Blood Transfusion)

 This system is created which aims to unify and standardize the manner of reporting of RBC
antigen.
 Machine and IO readable system of reporting red cell antigen was used

 ABO Blood group system: 001 : ABO

 For the 4 major antigen:
o Blood Type A = 001/ ABO1
o Blood Type B = 002/ ABO2
 o Blood Type AB = 003/ ABO3
o Blood Type O = 004/ ABO4
 Numerical System of ISBT
o Composed of 6 digits
 First 3 digits = ABO system
 Last 3 digits = antigen

o Blood Type A = 001001

o Blood Type B = 001002
o Blood Type AB = 001003
o Blood Type O = 001004

Blood Types / Genotypes based on parents’ alleles

Parent alleles A B O
A AA AB AO
(A) (AB) (A)
B AB BB BO
(AB) (B) (B)
O AO BO OO
(A) (B) (O)
Examples:

1. Parent blood type: OO x AO

o Possible blood type of children: 50% blood type A; 50% blood type O
o Exclusion: AB, B
2. Parent blood type: BO x AB
o Possible blood type of children: AB (AB), BB (B), AO (A),BO (B)
o 50% B ; 25% A ; 25% AB
o Exclusion: OO

ABO BLOOD TYPE FREQUENCIES

BLOOD TYPE ASIANS

O 40%
A 28%
B 25%
AB 7%
 THE FORMATION OF A, B, AND H ANTIGEN

 The formation of ABH antigen results from the interaction of genes at 3 separate loci (ABO, Hh,
Se)
 In as early as 37th day of fetal life, red cell antigens are already expressed on the RBC
o Antibodies develops more slowly than antigens
 ABO genes code not for the antigen themselves but for the production of glycosyltransferases
that add immunodominant sugars to a basic precursor substances (Paragloboside)

 Glycosyltransferase
o Catalyze the transfer of immunodominant sugar from the plasma to a basic precursor
substance
o Immunodominant sugars:
 Responsible for antigen specificity
 Basic Precursor Substance
o Also known as PARAGLOBOSIDE / GLYCAN
o Is an oligosaccharide structure composed of 4 interlinked sugars
o It is the acceptors structure on RBC for sugars L-fucose
 L-fucose = responsible for H antigen specificity
 *H antigen is always present because it serves as acceptor antigen structure
o 2 types:
1. Type 1
- Is the precursor substance for glycoprotein
- The antigens in blood group system is glycoprotein
2. Type 2
- Is the precursor substance for glycolipid

Gene Glycosyltransferases Immunodominant sugar Locus

H gene L-fucosyltransferase L-fucose Chromosome 19
(secretor locus)
A gene N-acetylgalactosaminyltransferase N-acetyl-D-galatosamine Long arm of Chromosome 9
B gene D-galactosyltransferase D-galactose Long arm of Chromosome 9

 For blood type AB, the 2 genes are present

 For blood type O, antigen is no formed.
o The H antigen structure is unmodified because it has no A and B antigen. It only
possesses the unmodified L-fucose
 Without the H structure, the A and B antigen are not present or expressed
 Rare blood group that does not have H antigen are Bombay (Oh) phenotype
 H gene is high frequency gene. This is present in 99.99% of all individual in the population.
o About 0.01% doesn’t have H gene which is known as H null/ H recessive type/ Bombay
(Oh) phenotype (hh)
 GENERAL CHARACTERISTIC OF ABH ANTIGEN

 On the 37th day of fetal life, attachment of immunodominant sugars occurs on the RBC
membrane and it is dependent on ABH genes inherited
 A antigen is weakly expressed on RBC during fetal life.
o A antigens only become strongly expressed a year after birth
 B and H antigen are strongly expressed on RBC during development
 May be found in secretions of people who are secretor. Secretor individuals are individuals who
inherit the so called “Secretor Gene”
o A Secretor individual (Sese+) can form the ABH soluble substances.
 These soluble substances are present in body secretion/fluid except in CSF
(present in all, especially saliva)
 CSF: contains blood-brain barrier = any big molecules cannot pass
o For nonsecretors (sese-), they do not form ABH soluble substances
 These antigens could also be found in bacteria and other species
 The ABH-like antigens may be found on the cell wall/surface of certain organism

INTERACTION OF THE Sese, Zz, and ABH GENE

 Secretor gene system (Sese) regulated the formation of H antigen and subsequently, of A and B
antigens in secretory cells.
o Therefore, it regulates the formation of glycoproteins (because of antigen B present in
the secretion)
 Zz system regulates production of H antigens on erythrocytes (glycolipids)

ABH ANTIGENS ABH Soluble substance

Red cells
In all body fluids except CSF
Epithelial tissues
Location **Blood brain barrier does not permit the
Bone marrow
entery of glycoproteins thus not seen in CSF
Other cell
Secreted Substances Glycolipid Glycoprotein
1st sugar in the precursor
Glucose N-acetylgalactosamine
substance (Paragloboside)
Type 1 and 2
Precursor chain Type 2 **type 1 is the predominant paragloboside for
the glycoproteins
1  4 linkage
Linkage
 Carbon of D-galactose to the carbon of N-acetylglucosamine
( pertains to interaction of the
 Type 2 precursor (1-3), first carbon of galactose to the 4th
carbon atom between sugars such
carbon of N-acetylglucosamine
as the carbon of D-galactose to the
For 1-3, first carbon of galactose to the 3rd carbon of N-
carbon of N-acetylglycosamine )
acetyglucosamine (FOR ABH SOLUBLE SUBS)
Regulating gene Zz gene Se gene
ABO ANTIBODIES

 Are mostly naturally occurring antibodies that are detectable 3 to 6 months after birth following
exposure to ABO-like antigens in the environment
 Are mostly IgM (cold reacting antibody) and react at room temperature or below
o 25C ( optimally at 4C )

GROUP ANTIGEN ANTIBODIES

O H Anti A
Anti B
Anti AB
**these 3 antibodies are mostly
of the IgG type.
**these antibodies are high
tittered clinically significant
A A, H Anti B
**mostly IgM high tittered
clinically significant
B B, H Anti A
**mostly IgM high tittered
clinically significant
AB A,B,H None

GENERAL CHARACTERISTICS OF ABO ANTIBODY

 These antibodies are not normally present at birth because they are developed 3-6 months after
birth
 In the neonatal serum, if ABO antibodies are detected, these antibodies present are not
considered to be the newborn’s antibody, it is maternally derived IgG antibody
o These antibodies were able to cross the placenta during pregnancy so these antibodies
coming from the mother can present in the newborn serum
 For the newborn, the only blood typing that should be done is Forward blood typing

Forms of ABO Antibody

1. Naturally Occuring
2. Immune Antibody

 For the immune antibodies, they are present in individuals of the following condition:
o 1. Patient with previous transfusion with mild ABO incompatibility
o 2. Female patient with previous pregnancy with mild ABO incompatibility
 The incompatibility is between the mother and the newborn due to the blood
type of the father
  ABO antibodies are present in some plants and animals
 They are present in low titer or even absent in some conditions such as:
o Patient with AIDS because these patients develop immunocompromised state and
therefore, have hypogammaglobulinemia
o Autoimmune disorder
o Active immune system (SLE, RA)
 The antibody of the people with this disease produces autoantibodies
 The remedy to this is immunosuppression to lower the antibody production
o Patient with Bruton’s disease (Congenital Gammaglobulinemia)
 They die of overwhelming infection because antibodies are not found

FORWARD / DIRECT TYPING

 Patient sample: RBC

BLOOD TYPE FORWARD TYPING

ANTI A ANTI B ANTI AB
( BLUE TYPING SERA ) ( YELLOW TYPING SERA )
O - - -
A + - +
B - + +
AB + + +

REVERSE / INDIRECT / BACKWARD TYPING

 Patient sample: serum

BLOOD TYPE REVERSE TYPING

A CELLS B CELLS O CELLS
O + + -
A - + -
B + - -
AB - - -
ABH SOLUBLE SUBSTANCES

 These are glycoproteins found in the secretion of secretor individuals (Sese) except CSF
 These are absent among nonsecretors (sese) individual
 Glycoproteins
 (+) secretors = inherit homozygous Secretor genes or heterozygous Secretor genes
o SeSe or Sese
 (-) secretors = inherit homozygous recessive secretors
o Sese

GENES BT/SECRETOR A SUB B SUB H SUB

 STAT
A, H, Se A type secretor None
B, H, Se B type secretor None
AB, H, Se AB type secretor
H, Se O type secretor None None

TEST FO ABH SOLUBLE SUBSTANCES:

SALIVA NEUTRALIZATION TEST

 Principle: Hemagglutination inhibition

 There is prevention of RBC clumping to occur
 Results:
o (+) = no agglutination
o (-) = agglutination
 2 step procedure:
o 1. Mix the saliva + antisera / lectin
o 2. Known RCS + mixture in step 1

STEP 1 STEP 2 REACTION ABH SOLUBLE SUBSTANCE

Saliva + Anti A Known A RCS No agglutination A substance
Saliva + Anti B Known B RCS No agglutination B substance
Saliva + Anti H Known O RCS No agglutination H substance

SAMPLE REACTION: AB SECRETOR

SALIVA + A CELL B CELL O CELL

Anti A (A sub) O O O
Anti B (B sub) O O O
Anti H (H sub) O O O

SAMPLE REACTION: B SECRETOR

SALIVA + A CELL B CELL O CELL

Anti A (A sub) 4+ O O
Anti B (B sub) O O O
Anti H (H sub) O O O