IMMUNOHEMATOLOGY
CHAPTER
ONE INTRODUCTON TO
IMUNOHEMATOLOYG
1.1 Overview of Immunohematology
Immuno hematology:
is more commonly known as "blood banking“
deals with the concepts and clinical techniques
related to modern transfusion therapy.
Is the area of laboratory medicine dealing with the
general procedures involved in collecting, preparing,
storing and transfusing blood.
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Immunohematology…
refersto immunologic reactions involving blood
components
an application of the principles of immunology to the
study of
red cell antigens and
their corresponding antibodies on blood for resolving the
problems of blood transfusions.
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1.2 Historical background
The first suggested case of ‘’transfusion “was the
rumored to have been given to pope innocent VIII in
July 1492.
The era of blood transfusion began when William
Harvey described the circulation of blood in 1616.
In 1665, Richard Lower, successfully performed the
first animal-to-animal blood transfusion.
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Historical background…
In 1667, jean Bapiste Denys transfused,
bloodfrom the carotid artery of a lamb into the vein of a
young man, which at first seemed successful.
using animal blood, but they were unsuccessful.
Later, it was found that it is impossible to
successfully transfuse the blood of one species of
animal into another species.
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Historical background…
Transfusions were prohibited from 1667 to 1818
Due to the disastrous consequences resulting.
In 1818, James Blundell of England successfully
transfused human blood to women suffering from
hemorrhage at childbirth.
Such species-specific transfusions seemed to work
sometimes but mostly the result was death.
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Historical background…
Karl Landsteiner
discovered the ABO blood groups in 1900,
introduced the immunological era of blood transfusion.
It became clear that the incompatibility of many
transfusion was caused by the presence of
certain factors on red (blood) cells now known as
antigens.
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Historical background…
Two main postulates were drawn:
1. Each species of animal or human have certain
factors on the red cells that are unique to that
species, and
2. Each species have some common and some
uncommon factors to each other.
This landmark event initiated the era of science
based transfusion therapy and was the foundation
of immunohematology as a science.
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1.3 Blood Group Genetics
Concerned with the way in which the different
blood groups are inherited
Chromosomes and Genes:
The nucleus of each human body cell contains 46 small
thread-like structures called chromosomes, arranged in
23 pairs.
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Blood Group ..
The length of each chromosome is divided into
many small units called genes.
Genes code for different inherited physical
characteristics, including blood groups
Blood Group Genetics…
Allomorphic genes (Alleles),and Polymorphism
Each gene has its own locus, along the length of
the chromosome.
Certain inherited characteristic can be represented
by a group of genes, and the locus can be
occupied by only one of these genes.
Such genes are called alleles or allomorphic genes.
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Blood Group Genetics…
Mitosis: While body cells multiply they do so by
producing identical new cells with 46
chromosomes.
Meiosis: When sex cells are formed either male
or female, the pairs of chromosomes do not
multiply but simply separate so that each of the
new cells formed contains only 23 chromosomes.
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Blood Group Genetics…
During fertilization when the egg and sperm
unite the fertilizer ovum receives 23
chromosomes from each sex cell.
Half of these from the male and
half from the female and thus will contain 46
chromosomes which arrange themselves in pairs in
the nucleus.
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Blood Group Genetics…
Genotype versus phenotype
Phenotype
Physicalexpression of inherited traits,
Determined by reacting red cells with known antisera
Genotype
Actualgenes inherited from each parent
Can only be inferred from the phenotype .
Family studies are required to determine the actual genotype .
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Table 1.1. The ABO phenotypes and their
corresponding genotypes
Phenotype Genotype
A AA, AO
B BB,BO
AB AB
O OO
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Blood Group Genetics…
Punnet square
Illustrates the probabilities of phenotypes from
known or inferred genotypes.
Visually portrays the potential offspring`s
genotypes or the probable genotypes of the
parents .
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Table1.2. Punnet squares showing ABO
inheritance
• Two group A parents can have a group O child.
• The parents of an AB child can be A, B or AB,
but not group O.
A O
A AA AO
O AO OO
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Fig 1: A sample family pedigree of three generations demonstrating
the basic Mendelian Inheritance of ABO blood groups.
* Make sure the students answer all the genotypes from the above
19 figure
1.3.1 Inheritance pattern of blood group Ags
In most cases blood group antigens are inherited
with co dominant expression.
The product of each allele can be identified when
inherited as a co dominant trait.
If
one parent passed on an A gene the other parent
passed on a B gene, both the A and B antigens would
be expressed equally on the red blood cells.
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Inheritance pattern…
Recessive or dominant inheritance patterns
Recessive
inheritance would require that the same alleles from both
parents be inherited to demonstrate the trait
Dominant
expression would require only one form of the allele to
express the trait.
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Inheritance pattern…
O gene is recessive, since it is expressed only
when both parents contribute the O allele.
The product of an O gene however, does not
affect the membrane proteins.
Itsexpression is termed as amorphic (a gene that
does not express a detectable product) rather than
recessive.
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Inheritance pattern…
Mendelian principles:
law of in dependen segregation
refers to the transmission of a trait in a predictable
fashion from one generation to the next.
Independent assortment is demonstrated by the fact
that blood group antigens inherited on different
chromosomes, are expressed separately and
discretely.
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1.3.2 Chromosomal assignment
Table 1.3.Chromosomal assignment of genes in blood
group system
Blood group system chromosome
Rh-----------------------------------------------------------1
Duffy-------------------------------------------------------1
Gerbich---------------------------------------------------2
MNS-------------------------------------------------------4
Kell---------------------------------------------------------7
ABO--------------------------------------------------------9
Kidd-------------------------------------------------------18
Lewis-----------------------------------------------------19
Landsteiner-Wiener-----------------------------------19
Lutheran-------------------------------------------------19
Hh---------------------------------------------------------19
P-----------------------------------------------------------22
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1.3.3 Homozygosity & Hetrozygosity
Homozygous
• Genotype is made up of identical genes, such as AA,
BB, or OO,
Heterozygous.
• Genotype is made up of different alleles from each
parent, such as AO, AB, or BO,
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Table 1.3.Dosage effect on Ag expression
Genotype Dosage effect on antigen
expression
Homozygous :MM Red blood cell tested with
anti-M : +4
Heterozygous :MN Red blood cell tested with
anti-M : +2
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1.3.4 Genetic inheritance
Genes can inherit with each other depending on
whether they are inherited on the:
same chromosome (Cis) or
opposite chromosome (Trans).
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Genetic inheritance…
Trans interaction may weaken the expression of
one of the antigens encoded by the genes,
For example:
The C and D genes of Rh system are inherited on
different genetic loci .
When C is inherited in trans to D, it will weaken the D
antigen expression on the red blood cell.
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Genetic inheritance…
Linkage and Haplotypes
In some blood group systems, the antigens are
encoded by two or more genes on the same
chromosome.
When genes are very close together, they are
inherited from each parent as a unit and are
known as linked
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Genetic inheritance….
Independent assortment does not occur when
genes are linked.
These gene units are called haplotypes
Silent genes
In some blood group systems genes do not
produce a detectable antigen product and are
called "silent” genes or amorphs.
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Genetic inheritance…
Amorphs can result in an unusual phenotype if
passed on by both parents.
The phenotypes are often called "null" type
because expressions of the blood group system
antigen are not apparent.
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Genetic inheritance…
Rare gene must be inherited from both parents
(homozygous) to produce a null phenotype.
null types caused by amorphic genes are rare.
Unusual phenotypes may also result from the
action of suppressors, or regulator genes.
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Genetic inheritance…
These genes (suppressor /regulator)
act to inhibit the expression of another gene and,
must be inherited in the homozygous state to create
this effect.
suppressor genes that affects the blood group antigen
are rare.
Null phenotype therefore can be resulted of either
an amorphic or a suppressor gene.
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Table 1.4.Blood group genes that can
result an unusual phenotypes
Blood group Amorph/Regulator Phenotype
system gene
H h Bombay
Rh r/x0r Rh null
Kell K0 Kell null
Lutheran Lu/in(lu) Lu(a-b-)
Kidd JK JK(a-b-)
Duffy Fy Fy(a-b-)
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Secretors and Non-Secretors
The term secretor and non-secretor only refer to the
presence or absence of water- soluble ABH antigen
substances in body fluids (saliva, semen, urine, sweat,
tears, etc).
Every individual contains alcohol soluble antigens in
body tissues and on the red cells .
Secretors possess the water soluble (glycoprotein)
form of antigen, which appears in most body fluids
Conti…
Majority of the population secrete water- soluble
substances in saliva and most other body fluids that
have the same specificity as the antigens on their red
cells.
The production of A, B & H antigens in saliva is
controlled by a secretor gene, which is in herited
independently of the ABO and H genes.
Cont…
The relevant gene is called Se, and its allele which
amorphic is se.
At least one Se gene (genotype SeSe or Sese) is
essential for the expression of the ABH antigens in
secretors.
Individual who are homozygous for se (sese) do not
secrete H,A, or B antigens regardless of the presence
of H,A or B genes
Cont…
The Se gene does not affect the formation of A,B or
H antigens on the red cells or in hematopoietic tissue,
which are alcohol soluble and which are not present
in body secretions.
Oh (Bombay) individuals do not secrete A, B or H
substance, even when the Se gene is present
1.4 Blood cell antigens
1.4.1 Red blood cell antigens
A unique set of red blood cell Ag is determined
through genetic inheritance.
These antigens protrude from the surface of the
RBC in three dimensional configurations.
As a result, they are accessible to Ab molecules for
agglutination reaction.
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Red cell antigens…
In biochemical terms these antigens may take the
form of:
proteins,
Glycoprotein,
Glycolipids
Some of the red blood cell antigens are more
immunogenic than the others
Example
The D antigen within the Rh group system.
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1.4.2 Human leukocyte antigens (HLA)
Is possessed by nucleated cells such as
leukocytes and tissues
Can readily provoke an immune response if
transferred in to a allogenic individual.
Encoded by genes which are parts of Major
Histocompatibility Complex (MHC) gene system.
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Human Leukocyte antigens…
The MHC system is important in the:
recognition of non self ,
coordination of cellular and humoral immunity , and
graft rejection .
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Human Leukocyte antigens…
The MHC region is on chromosome 6 and is
divided in to three categories or classes :
Class I includes the A, B and C locus,
Class II includes the DR, DP and DQ
Class III includes the complement proteins
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Human Leukocyte antigens…
The MHC region is called polymorphic , because
there are so many possible alleles .
For example :
At least 49 different alleles or possible genetic expressions
have been identified at the A locus.
At the B locus 97 alleles are identified.
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1.4.3 Platelet antigens
Platelet possesses inherited membrane proteins
that can also elicit an immune response.
Platelet antibodies are less frequently found,
because there is less antigen variability in the
population.
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Platelet antigens…
Antibodies to platelet antigens are the major
cause of :
neonatal alloimmune thrombocytopenia,
Post transfusion purpura ,
Itcan also decrease the expected increment of
platelet transfusion.
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1.5 Blood group Abs & their stimulation
Blood group antibodies are classified into:
Natural and
Immune antibodies.
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1.5.1. Natural / non red cell immune Abs
Are RBC Abs in the serum of an individual that
are not provoked by previous RBC sensitization.
The term non red cell immune have crept in to
modern use.
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Non-red cell immune antibodies…
Characteristics
They are mainly IgM type.
Exhibit optimum in vitro agglutination saline media:
complete antibodies.
Optimum reaction at room temperature or lower:
cold agglutinins.
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Non-red cell immune antibodies…
Do not react above the body temperature
most of these do not give rise to transfusion reactions.
They are of high molecular weight
cannot cross the placenta
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1.5.2. Immune antibodies
Produced due to previous antigenic stimulation
either by transfusion or pregnancy
Characteristics
Mainly IgG type
Do not exhibit visible agglutination in saline, but
in albumin medium: Incomplete antibodies.
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Immune antibodies…
Optimally react at 370C: warm agglutinins.
Causes more serious transfusion reactions than
the naturally occurring ones.
Can cross the placental barrier.
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1.6 Antigen - Antibody interactions
The binding follow the law of mass action and
is a reversible process.
This union complies with the principles of a
chemical reaction that has reached equilibrium.
When Ag and Ab combines, an immune complex
is produced.
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Ag-Ab interactions…
The amount of Ag - Ab complex formation is
determined by the association constant of the
reaction .
When the forward reaction rate is faster than the
reverse reaction rate Antigen-Antibody complex
formation is favored.
Therefore a higher association constant influences
greater immune complex formation at equilibrium
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Ag-Ab interactions…
Properties that can influence the binding of Ag
and Ab
The goodness of fit (as a lock and key fit)
complementary nature of the antibody
size, shape, and charge of antigen
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