ABO Blood Group System

Renee Newman Wilkins, PhD, MLS(ASCP)cm

CLS 325/435

School of Health Related Professions

University of Mississippi Medical Center

History: Karl Landsteiner

Discovered the ABO Blood Group System in 1901

He and his five co-workers began mixing each others red cells and serum together and inadvertently performed the first forward and reverse ABO groupings http://www.nobelpreis.org/castellano/medizin/images/landsteiner.jpg

Why is it important?

ABO compatibility between donor cell and patient serum is the essential foundation of pretransfusion testing

It is the only system with expected antibodies Whether they are IgG or IgM, ABO antibodies

can activate complement readily This means that incompatibilities can cause life

threatening situations (transfusion reactions)

ABO antigens:

Biochemical & Genetic Considerations

ABO and H Antigen Genetics

Genes at three separate loci control the occurrence and location of ABO antigens

The presence or absence of the A, B, and H antigens is controlled by the H and ABO genes

Location

The presence or absence of the ABH antigens on the red blood cell membrane is controlled by the H gene

The presence or absence of the ABH antigens in secretions is indirectly controlled by the Se gene

ABO Antigen Genetics

H gene – H and h alleles (h is an amorph)

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

ABO genes – A, B and O alleles

H Antigen

The H gene codes for an enzyme that adds the sugar fucose to the terminal sugar of a precursor substance (PS)

The precursor substance (proteins and lipids) is formed on an oligosaccharide chain (the basic structure)

RBC Precursor Structure

Glucose

Galactose

N-acetylglucosamine

Galactose

Precursor Substance (stays the

same)

RBC

Formation of the H antigen

Glucose

Galactose

N-acetylglucosamine

Galactose

H antigen

RBC

Fucose

H antigen

The 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 Immunodominant sugars are present at the

terminal ends of the chains and confer the ABO antigen specificity

A and B Antigen

The “A” gene codes for an enzyme (transferase) that adds N-acetylgalactosamine to the terminal sugar of the H antigen N-acetylgalactosaminyltransferase

The “B” gene codes for an enzyme that adds D-galactose to the terminal sugar of the H antigen D-galactosyltransferase

Formation of the A antigen

Glucose

Galactose

N-acetylglucosamine

Galactose

RBC

FucoseN-acetylgalactosamine

Formation of the B antigen

Glucose

Galactose

N-acetylglucosamine

Galactose

RBC

FucoseGalactose

Genetics

The H antigen is found on the RBC when you have the Hh or HH genotype, but NOT from the hh genotype

The A antigen is found on the RBC when you have the Hh, HH, and A/A, A/O, or A/B genotypes

The B antigen is found on the RBC when you have the Hh, HH, and B/B, B/O, or A/B genotypes

H antigen

Certain blood types possess more H antigen than others:

O>A2>B>A2B>A1>A1BGreatest

amount of HLeast

amount of H

Why do Group O individuals have more H antigen than the other groups?

Group O individuals have no A or B genes to convert the H antigen to A or B antigens….that means more H antigen sites

Group O Group A

Many H antigen sites

Fewer H antigen

sites

A

A A

AA

Most of the H antigen sites in a Group A individual have been

converted to the A antigen

ABO Antigens in Secretions

Secretions include body fluids like plasma, saliva, synovial fluid, etc

Blood Group Substances are soluble antigens (A, B, and H) that can be found in the secretions. This is controlled by the H and Se genes

Secretor Status

The secretor gene consists of 2 alleles (Se and se)

The Se gene is responsible for the expression of the H antigen on glycoprotein structures located in body secretions

If the Se allele is inherited as SeSe or Sese, the person is called a “secretor” 80% of the population are secretors

Secretors

Secretors express soluble forms of the H antigen in secretions that can then be converted to A or B antigens (by the transferases)

Individuals who inherit the sese gene are called “nonsecretors” The se allele is an amorph (nothing expressed) sese individuals do not convert antigen precursors

to H antigen and has neither soluble H antigen nor soluble A or B antigens in body fluids

Secretor Status Summary

The Se gene codes for the presence of the H antigen in secretions, therefore the presence of A and/or B antigens in the secretions is contingent on the inheritance of the Se gene and the H gene

Se gene (SeSe or Sese)

H antigen in secretions

A antigen

B antigen

se gene (sese)

No antigens secreted in saliva or other

body fluids

and/or

ABO GroupABH

Substances

Secretors (SeSe or Sese): A B H

A +++ 0 +

B 0 +++ +

O 0 0 +++

AB +++ +++ +

Non-secretors (sese):

A, B, O, and AB 0 0 0

Sese + h/h (no H antigen) no antigens in secretions

Lewis (Le)

The Lewis Blood Group System is mentioned here because it is related to secretor status

Lewis antigens are plasma antigens formed by tissues and are released into plasma where they adsorb onto the RBCs (they are not an integral part of the RBC membrane)

Consists of 2 antigens Lea

Leb

Lewis

Lea and Leb are a single gene (Le) and its amorph (le) Lea is a precursor to Leb

The Le gene codes for a transferase, which attaches L-fucose to the precursor chain to form the Lea antigen (designated Le(a+b-)

If the H and Se genes are inherited, the Lea is converted to Leb and is designated Le(a-b+)

In childhood, both may be on the RBC, Le(a+b+) If a person is lele, they will have no Lewis

antigens in plasma or on red blood cells

Frequency of Lewis phenotypes*

Phenotype Whites Blacks

Le(a+b-) 22% 23%

Le(a-b+) 72% 55%

Le(a-b-) 6% 22%

lele

*Adapted from Flynn, J. (1998). Essentials of Immunohematology

ABO Subgroups

ABO subgroups differ in the amount of antigen present on the red blood cell membrane Subgroups have less antigen

Subgroups are the result of less effective enzymes. They are not as efficient in converting H antigens to A or B antigens (fewer antigens are present on the RBC)

Subgroups of A are more common than subgroups of B

Subgroups of A

The 2 principle subgroups of A are: A1 and A2

Both react strongly with reagent anti-A To distinguish A1 from A2 red cells, the lectin

Dolichos biflorus is used (anti-A1)

80% of group A or AB individuals are subgroup A1

20% are A2 and A2B

A2 Phenotype

Why is the A2 phenotype important? A2 and A2B individuals may produce an anti-A1

This may cause discrepancies when a crossmatch is done (incompatibility)

What’s the difference between the A1 and A2 antigen? It’s quantitative The A2 gene doesn’t convert the H to A very well The result is fewer A2 antigen sites compared to

the many A1 antigen sites

A1 and A2 Subgroups*

Anti-A antisera

Anti-A1 antisera

Anti-H lectin

ABO antibodies in serum

# of antigen sites per

RBC

A14+ 4+ 0 Anti-B 900 x103

A24+ 0 3+ Anti-B &

anti-A1

250 x103

*Adapted from Flynn, J. (1998). Essentials of Immunohematology

Other A subgroups

There are other additional subgroups of A Aint (intermediate), A3, Ax, Am, Aend, Ael, Abantu

A3 red cells cause mixed field agglutination when polyclonal anti-A or anti-A,B is used

Mixed field agglutination appears as small agglutinates with a background of unagglutinated RBCs

They may contain anti-A1

B Subgroups

B subgroups occur less than A subgroups B subgroups are differentiated by the type of

reaction with anti-B, anti-A,B, and anti-H B3, Bx, Bm, and Bel

Other ABO conditions

Bombay Phenotype (Oh) Inheritance of hh The h gene is an amorph

and results in little or no production of L-fucosyltransferase

Originally found in Bombay (now Mumbai)

Very rare (130 worldwide)

Bombay

The hh causes NO H antigen to be produced Results in RBCs with no H, A, or B antigen

(patient types as O) Bombay RBCs are NOT agglutinated with

anti-A, anti-B, or anti-H (no antigens present) Bombay serum has strong anti-A, anti-B and

anti-H, agglutinating ALL ABO blood groups What blood ABO blood group would you use

to transfuse this patient??

ANSWER:

Another Bombay Group O RBCs cannot be given because they still

have the H antigen You have to transfuse the patient with blood that

contains NO H antigen

ABO Blood Group:

ABO Antibodies

Landsteiner’s Rule:

Normal, Healthy individuals possess ABO antibodies to the ABO antigen absent from their RBCs

ABO Blood Group System

The ABO Blood Group System was the first to be identified and is the most significant for transfusion practice

It is the ONLY system that the reciprocal antibodies are consistently and predictably present in the sera of people who have had no exposure to human red cells

Blood Group Systems

Most blood group systems (ABO and others) are made up of: An antigen on a red cell and the absence of it’s

corresponding antibody in the serum (if you’re A, you don’t have anti-A)

If you do NOT have a particular antigen on your red cells then it is possible (when exposed to foreign RBCs) to illicit an immune response that results in the production of the antibody specific for the missing antigen

ABO

Remember: The ABO Blood Group System does NOT require

the presence of a foreign red blood cell for the production of ABO antibodies

ABO antibodies are “non-red blood cell stimulated” probably from environmental exposure and are referred to as “expected antibodies”

ABO antibodies

group A serum contains anti-B group B serum contains anti-A group AB serum contains no antibodies group O serum contains anti-A, anti-B, and

anti-A,B

Anti-A1

Group O and B individuals contain anti-A in their serum

However, the anti-A can be separated into different components: anti-A and anti-A1

Anti-A1 only agglutinates the A1 antigen, not the A2 antigen

There is no anti-A2.

Anti-A,B

Found in the serum of group O individuals Reacts with A, B, and AB cells Predominately IgG, with small portions being

IgM Anti-A,B is one antibody, it is not a mixture of

anti-A and anti-B antibodies

ABO antibodies

IgM is the predominant antibody in Group A and Group B individuals Anti-A Anti-B

IgG (with some IgM) is the predominant antibody in Group O individuals Anti-A,B (with some anti-A and anti-B)

ABO antibody facts

Reactions phase: Room temperature Complement can be activated with ABO

antibodies (mostly IgM, some IgG) High titer: react strongly (4+)

ABO Antibodies

Usually present within the first 3-6 months of life

Stable by ages 5-6 years Decline in older age Newborns may passively acquire maternal

antibodies (IgG crosses placenta) Reverse grouping (with serum) should not be

performed on newborns or cord blood

Nature of antibodies

Non-red blood cell stimulated (previously discussed) ABO antibodies

Red blood cell stimulated Antibodies formed as a result of transfusion, etc Usually IgG Active at 37°C Can occur in group O (may occur in group A or B) These antibodies also occur in the other Blood Group

Systems

Laboratory Testing:

ABO typing

ABO Blood Groups

ABO Group

Antigen Present

Antigen Missing

Antibody Present

A A B anti-B

B B A anti-A

O None A and B anti-A, anti-B, anti-A,B

AB A and B None None

Forward Grouping

Reaction of patient red blood cells tested with Reagent anti-A and anti-B antisera

Reverse Grouping

Reaction of patient serum with reagent Group A and Group B cells

Forward & Reverse Typing

anti-A anti-B A cells

B cells

ABO group

% US white pop.

% US black pop.

1 0 0 + + O 45 49

2 + 0 0 + A 40 27

3 0 + + 0 B 11 20

4 + + 0 0 AB 4 4

Reaction of cells tested with:

Reaction of serum tested with: