23-Sep-14 1

BLOOD GROUPS
Mansyur Arif
Dept. of Clinical Pathology, Faculty of
Medicine, Hasanuddin University /
Dr. Wahidin Sudirohusodo Hospital
Makassar
23-Sep-14 2
Definitions
Blood groups are determined by antigens
structures on the surfaces or red cells and
are detected by reactions with specific
antibodies.
A blood group system is defined by a.g that
are regulated either by allelic genes or
closely linked genes.
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The number or red cell blood groups now exceeds 400.
(table1).
Table 1. Survey of major Red Cell Blood Group System
System Important antigens
ABO A
1
,A
2
,B,H,A
3
,A
m
,A
x

MNSs M,N,S,s,U,M
g
,Mi
a
,Hu,HeMt
a
,Vw,M
2
,N
2
,S
2

P P
1
,p
k
,P2,(Tj
a
)
Rh D,C,E,c,e,C
w
,E
w
,ce,Ce,G,CE,cE,D
u
,C
u
,E
u
,LW
Lutheran Lu
a
,Lu
b

Kell K,k,Kp
a
,Kp
b
,Js
a
,Js
b
Lewis Le
a
,Le
b

Wright Wr
a
,Wr
b

Diego Di
a
,Di
b
Cartwright Yt
a
,Yt
b
Xg Xg
a

Dombrock Do
a
,Do
b

Colton Co
a
,Co
b
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Antibodies : sources & properties
1. Normal humans
A.bodies to some blood group a.g occur in
the serum of individuals who lack the a.g
and have had no prior exposure to it
natural isohemagglutinins.
The major ones are directed against surface
a.g such as the ABO, Ii and P systems
controlled by oligosaccharides
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Isohemagglutinins with ABO are always
clinically significant
Isohemagglutinins elicited by similar
sequences on microbial surfaces >>Ig Ms
effective hemolysins because they
efficiently fix complement.
Occasionally Ig G a.bodies specific for these
a.g also appear.


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2. Immunized animals
If animals are immunized with human red cells
may form a.bodies to certain of the
xenogeneic blood group a.g important
source of blood group anti sera carefully
absorbed with human red cells to establish
specificity.
Recently developed a.g specific monoclonal
a.bodies do not require such absorption.

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3. Immunized humans
The third major source of the blood group anti
bodies are donors who have been allogenically
immunized either by (1) prior blood
transfusions or (2) previous pregnancies
immune antibodies elicited by prior
exposure to red cell a.g are commonly IgGs

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Methods of detection
1. Agglutination by specific antibody
Under physiologic conditions of pH and ionic
strength, normal red cells repel each other
owing to their negative surface charge or
zeta potential
2. Enhancement of agglutination by antibody
a. Reduction of zeta potential
Can be reduced by addition of colloid (alb,
polyvinylpyrrolidone or dextran).
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b. Insertion of a.b red cells bridges
Agglutinations may produced or enhanced
by addition of Coomb reagent (i.e.,anti-
globulin a.body)
3. Use of lectins
4. Automated techniques



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Genetics
According to Mendelian laws
Heredity is generally autosomal
codominant i.e there is an expression of
both alleles in the heterozygous individual
1.Linked genes
2.Interaction with other genes
3.Loci of blood groups genes on
chromosomes (table 2)
4.Occurrence of blood group antigens

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Table 2. Chromosome Assignment of Some Blood
Group Loci

Locus Chromosome

ABO 9
Rh 1
Fy 1
Chido, Rogers 6
MNSs 4
Xg X
Sc 1
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ABO SYSTEM
a. Historical notes
In subsequent work Landsteiner recognized
that the pattern of reactions could be
explained by two a.g, which designated A
and B. O signified the state of not having A
or B.
Table 3. The Landsteiner scheme
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Table 3. The ABO system defined by Anti-A and Anti B

Blood Groups Antigens on RC Antibodies in serum
O
A
B
AB
None
A
B
A and B
Anti-A and Anti-B
Anti-B
Anti-A
None
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b. Subdivisions of A antigen
A antigen and anti-A are complex. Anti-A
serum from a group B donor contains 2 types
of a.b, anti-A and anti-A
1 .
(table 4)

Group Antigens
Reaction with
Anti-A Anti-A
1
A
1
A
2
AA
1
A
+
+
+
-
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Genetics
Determining the blood group : genotype and
phenotype. A child receives one of four genes
from each parent : A
1
, A
2
, B, or O. Six
phenotypes are possible because the A a.g
associated with group A
2
and also A
1.
There are ten possible genotypes. Group A
1
may
have 3 genotypes (A
1
A
1
, A
1
O, A
1
A
2
). Group A
2
can have either A
2
A
2
or A
2
O genotypes. Group
B can have either BB or BO genotypes
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Genotype :
- specific genes that person carries
- determined by family studies
- AA, AO, BB, BO, AB and OO
- see fig 1.

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Family 1


Phenotype B A
1
Genotype BO A
1
O



Phenotype O O A
1
B
Genotype OO OO A
1
B

Family 2


Phenotype A
1
A
2
B
Genotype A
1
A
2
A
2
B


Phenotype A
2
B A
1
Genotype A
2
B A
1
A
2

Fig 1. Illustration of usefulness of family studies in the elucidation of phenotype





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Phenotype :
Four phenotypes : A, B, AB and O
Although there are ten possible genotypes,
the absence of a specific anti-O prevents the
serological recognition of more than four
phenotypes. (table 5)
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Table 5. Blood Type
Phenotype Genotype Antigens
on red cell
Antibodies in
plasma
O
A
B
AB
OO
AA or AO
BB or BO
AB
O
A
B
AB
Anti-A, Anti-B
Anti-B
Anti-A
None
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Fig.2 Synthesis of ABH antigens

R

glc gal glcnac gal
H precursor

Hh gen



fuc
R

glc gal glcnac gal H antigen





A gene B gene



fuc fuc
R R

glc gal glcnac gal glcnac glc gal glcnac gal gal
A antigen B antigen
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H antigen
The surfaces oligosaccharides that
constitutes the H a.g is the precursor of the
A and B a.g
Gene A & B responsible for converting H
substance into A & B substance
The O gene is an amorph and doesnt
transform the H substance
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Rare variant Bombay, the H precursor
cannot be converted to H lack H ag &
hence A or B phenotype cant be expressed.
A terminal sugar molecules determine a.g
specificity :
A a.g : N acetylgalactosamine
B a.g : galactosa

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Other Carbohydrate Antigen
a. Lewis system
The Lewis a.g are made from the same
precursors as the ABH a.g except that they
are exclusively type 1 chain.
The expression ag depends on the
interaction of the H gene, Se gene and Le
gene
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b. P system
These ag were recognized by antisera
developed in rabbits glycosphingolipids
and originate on a ceramide dihexose (Gal-
Gal-ceramide)
c. Ii system
Most cold a.bodies have specificity against
the Ii a.g system. These a.g are found in red
cells and nonhematopoietic tissue

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Rhesus System
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Rhesus a.b >> immune (previous
transfusion or pregnancy), naturally <<
Anti-D is responsible for most of the clinical
problems associated with the system the
simple subdivision of subjects into Rh D +
and Rh D , using anti-D is sufficient for
routine clinical purposes.


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A. Nomenclature : relation to genetic models

1. Fischer-Race theory (table 6) :
Postulates 3 closely linked genes Cc,
Dd and Ee. Rhesus a.g is renamed D.
Rhesus positive presence of the D
antigen, also called Rh or Rh factor
Rhesus negative absence of D but
doesnt denote absence of other a.g
of the Rh system (C,c,E or e)

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2. Weiner system
3. Rosenfield system
B. Compound antigens
C. Weakened antigens :
- weakly reactive ag D
u

- formal terminology : Rh
+
, D
u
variant
- for transfusion : D
u
is equivalent to

Rh
+


D. Deleted antigens : Rh null cells.
E. Rh antigens structure
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Table 6. Rh gene complexes
Fischer-Race Wiener

CDe R
1

cde r
cDE R
2
cDe R
o
C
w
De R
1W
cdE r
u
Cde r
1
CDE R
z
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Other clinically significant systems
1. Kell system
The Kell a.g system rivals the Rh system in its
complexity and clinical importance. Appearing in
response to prior immunization, anti-Kell a.b have
caused hemolytic transfusion reactions and HDN.
The main a.g pairs : K-k, Kp
a
-Kp
b
and Js
a
-Js
b
2. Duffy system
Double negative phenotype red cells, Fy (a-b-) are
totally resistant to invasion by Plasmodium vivax.
Transfusion of incompatible blood into Duffy-
sensitive individuals can cause severe hemolysis.
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3. Kidd system
Immunization to Kidd is caused mainly by
transfusions. Kidd a.b are evanescent warm-active
incomplete a.b that may not be detected in red cell
a.b screens. Consequently they often cause
delayed transfusions rx, which may be severe.
4. Lutheran system
There are 2 common alleles, Lu
a
and Lu
b
and a
silent one. The double-negative phenotype caused
by either dominant inhibitor gene or a recessive
silent allele.
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5. Xg
a
blood group
This a.g is controlled by a gene on the X
chromosome. Its not clinically significant but
is of interest as a marker for X chromosome
that appear to escape inactivation by the
Lyon mechanism.
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The ABO and Rhesus (Rh) groups are of major clinical
significance. Some other systems of less overall
importance are listed in table 7.
Systems Frequency of a.body Cause of HDN

ABO Very common Yes
Rh Common Yes
Kell Occasional Yes
Duffy Occasional Yes
Kidd Occasional Yes
Lutheran Rare No
Lewis Occasional No
P Occasional Yes (rare)
MN Rare Yes (rare)
Ii Rare No




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Uses of blood grouping data
A. In clinical medicine
1. Pretransfusion testing :
Prior to transfusion, blood is typed
and crossmatched to establish ABO and
D compatibility
2. Hemolytic disease of the newborn
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B. In genetics chromosome mapping
C. In forensic medicine :
1. Identification studies
2. Paternity testing
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Thank you