Chapter 6 These genes produce specific

The ABO Blood Group System glycosyltransferases that add sugars to a basic
precursor substance.
Routine ABO Testing
Forward grouping Importance of the H antigen
Reverse grouping 1. Production of A, B, and H antigens in fetal and
Routine reagents for ABO testing adult life
Requirements for routine ABO testing Interaction of Hh and ABO Genes
Inverse reciprocal relationship between the Immunodominant sugars confer blood group
forward and reverse type specificity
Naturally occurring antibodies <Insert Figure 64>
Postulated role of bacteria Importance of the H gene
ABO Antibodies Bombay phenotype
Predominantly IgM Importance of the A gene
Activate complement Importance of the B gene
Room temperature or colder <Insert figures 6-5 and 6-6>
Produce strong direct agglutination reactions Considerations for the AB blood group
Reagent anti-A
Reagent anti-B Molecular Genetics of ABO
Reagent anti-A,B Investigation of ABO alleles, epitope
Monoclonal antisera structures, and exons
Polyclonal antisera ABO gene located on chromosome 9 and
Considerations for routine blood bank consists of seven exons
testing Diversity at the ABO locus

Inheritance of the ABO Blood Groups

Work of Bernstein Formation of A, B, and H
Inheritance by Mendelian genetics Soluble Antigens
Codominant expression ABH antigens are integral parts of the
membranes of various cells
ABO Genotypes and Phenotypes ABH-soluble antigens can be found in all
body secretions
<Insert Figure 6-8>
Genotype Phenotype Presence in secretions is dependent on ABO
A1A1 A1 and secretor genes inherited.
A1A2 A1 Nonsecretors are possible.
A1O A1
A2A2 A2
A2O A2
A1B A1B
A2B A2B
OO O
BB B
BO B
Formation of A, B, and H ABO Subgroups
Red Cell Antigens Phenotypes that show weaker variable
ABH antigen formation results from the serological reactivity with the commonly
interaction of genes at three separate loci used human polyclonal Anti-A, Anti-B, and
(ABO, Hh, and Se). Anti-A,B reagents.
 Monoclonal typing reagents used routinely.
A Subgroups
First described in 1911 by von Dungen
A1 and A2 subgroups
Differences between A1 and A2 are
quantitative and qualitative
A subgroups generally more common than B
subgroups
Differences in conversion of H precursor
substance among A subgroups
Effects of polymorphism at the ABO locus
Forward grouping reagent (Anti-A) strongly
agglutinates both A1 and A2 phenotypes
Anti-A1 lectin reagent used in the
differentiation of A1 and A2 phenotypes
Anti-A1 lectin reagent agglutinates A1 (or A1B)
cells but does not agglutinate A2 (or A2B cells)
Lectins used in blood banking
Dolichos biflorus: agglutinates A1 or A1B
Bandeiraea simplicifolia: agglutinates B cells
Ulex europaeus: agglutinates O cells (H specificity)
and other ABO blood groups depending on the
amount of H antigen available
Varying concentration of H antigen among
blood groups
Anti-H is occasionally found in the serum
A naturally occurring IgM cold agglutinin that
reacts best below room temperature
Possible problems in antibody screening
procedures
Use of Anti-H lectin
Weak A Subgroups
Subgroups weaker than A2 occur infrequently
Most often recognized through an ABO
discrepancy (unexpected reactions in the
forward and reverse grouping )
Varying expression of four characteristics
Weak A subgroups can be distinguished as A3,
Ax, Aend, Am, Ay, and Ael
Weak B Subgroups
Very rare and less frequent than A subgroups
Usually recognized by variations in the
strength of the reaction using anti-B and anti-
A,B
Result of alternate alleles at the B locus
Five criteria used for differentiation of weak B
phenotypes
Serologic techniques characterize B subgroups
in the following categories: B3, Bx, Bm, and Bel
The Bombay Phenotypes (Oh)
First reported by Bhende in 1952 in Bombay,
India
Inheritance of a double dose of the h gene,
producing the very rare genotype hh
No H antigen made
ABO genes cannot be expressed
ABH antigens cannot be formed
RBCs are devoid of normal ABH antigens and
fail to react with anti-A, anti-B, and anti-H.
In RBC testing using anti-A and anti-B, the
Bombay would phenotype as an O blood
group.
Unlike the anti-H found occasionally in the
serum of A1 and A1B individuals, the
Bombay anti-H can often be potent and reacts
strongly at 37C .
It is an IgM antibody that can bind
complement and cause RBC lysis.
Considerations for transfusions
Only blood from another Bombay individual
will be compatible
Underlying molecular defect of the Bombay
phenotype
The Para-Bombay Phenotypes
Rare phenotypes in which the RBCs are
completely devoid of H antigens or that have
small amounts of H antigen present
ABH Antigens and Antibodies in Disease
Associations between ABH antigens and
leukemias demonstrating
hypogammaglobulinemia, such as CLL
Other leukemias with chromosome 9
translocations
Any hemolytic disease inducing stress
hematopoiesis (e.g., thalassemia)
"Acquired B" phenomenon in group A1
individuals
 ABO Discrepancies
Unexpected reactions in the forward and
reverse grouping due to
Problems with the patients serum (reverse
grouping)
Problems with the patients red cells (forward
grouping)
Problems with both the serum and cells
Can appear as extra positive or weak/missing
reactions
Must be resolved prior to reporting a patient
or donors ABO group

Categories of ABO Discrepancies

Group I discrepancies
Group II discrepancies
Group III discrepancies
Group IV discrepancies

Resolution of Common Group II Discrepancies

Enhancing weakly reacting antigens with room
temperature incubation
Pretreatment of RBCs with enzymes
Acquired B antigen

Resolution of Common Group III Discrepancies

Effects of Rouleaux
Effects of Whartons jelly

Resolution of Common Group IV Discrepancies

Consideration of cold autoantibodies
The patients RBCs can be tested with Dolichos
biflorus to confirm the presence of an ABO
subgroup
Unexpected alloantibodies in the patients serum
other than ABO isoagglutinins may cause a
discrepancy in the reverse grouping