Immunogenetics

 Immunogenetics is the study of the mechanisms of

autoimmune diseases, tolerance in organ transplantation,
and immunity to infectious diseases—with a special
emphasis on the role of the genetic make-up of an
organism in these processes.
 Historically, the launch of immunogenetics could be
traced back to the demonstration of Mendelian inheritance
of the human ABO blood groups in 1910.
 The most important influence on the development of
immunogenetics is the studies of the MCH gene family.
1. The genetic basis of antibody
structure
 Introduction
 How can the immune system recognize an almost unlimited
number of foreign antigens?
 Remarkably, each mature lymphocyte is genetically
programmed to attack one and only one specific antigen:
each mature B cell produces antibodies against a single
antigen, and each T cell is capable of attaching to only one
type of foreign antigen.
 Question: Every person has the ability to generate 1015 to
1018 different Ig or TCR, since genomes of many species
have only 30000-40000 gene, how do so few genes
produce so many different antigen receptor molecules?
 The investigators over the last 30yrs, found that V
and C regions of Ig are coded for by different genes
—many different V region genes that can be linked
up to a single C region gene.

 Susumu Tonegawa found that antibody genes can

move and rearrange themselves within the genome
of a differentiating cells: a variable (V) region gene
can be located in one position in the DNA of an
inherited chromosome but then move to another
position on the chromosome during lymphocyte
differentiation.
 Review
 Genome: linear arrays of genes in the DNA strands of the various
xmes.
 Every diploid cell in the human body contains the same set of
genes as every cell—except the lymphocytes which differ from
other cells and each other in the content of genes coding for their
antigen-specific receptor.
 The expression of a specific pattern of genes determines the cell’s
function. E.g.: only pancreatic  cells express Insulin, only B cells
express antibodies.
 Most of genes coding for a protein have exons and introns (spliced
out during RNA processing).
 Genes coding for cell surface proteins have a leader sequence (L
exon) at the 5’end, coding for signal peptide.
General gene structure and gene
expression
Genetic events in the synthesis of Ig chains
1. VL is coded for by two separate gene segments: a variable
(V) segment, which codes for the amino-terminal 95
residues.
2. And a small joining (J) segment, codes for about 13 (96-
108) residues at the carboxy-terminal end of the variable
region.
3. One V gene and one J gene are brought together in the
genome to create a gene unit that, together with the C
region gene, codes for an entire Ig L chain. This unique
gene rearrangement mechanism-refered to as V(D)J
recombination, is used only by genes coding for IgL and H
chains and genes coding for TCRs.
V J C
The genetic events leading to the synthesis of
a kappa light chain

(On chromosome 2)
 An enzyme complex, known as V(D)J recombinase,
mediates the rearrangement of receptor genes in B
and T cells. RAG-1 and RAG-2 (recombination-
activating genes) are required in the first stages of
cutting Ig- and TCR-DNA.
 There are approximately 40 different V genes, and 5
J genes.
  Chain synthesis
 The  genes are found on chromosome 22 in the
human.
 The synthesis of  chains is similar in principle to the
synthesis of  chains in that it involves rearrangement
of DNA, which joins a V  gene with a J  segment.
 There are about 40 V , 4 J  and 4 C .
Organization and rearrangement Heavy chains
 In contrast to the variable region of a light chain that is
constructed from 2 gene segments, the variable region of a
heavy chain is constructed from 3 gene segments (VH, DH
(diversity segment) & JH). The D and J segments code for
aa sequences in the 3rd hypervariable, or complementarity
determining region (CDR3) of the heavy chain.
 The second feature of the H chain genes is the presence in
the germline of multiple genes coding for the C region of
the Ig.
Five different classes: , , , , and .
Immunoglobulin heavy chain isotypes
Isotype Heavy Chain
IgM 
IgD 
IgG 
IgA 
IgE 
IgG IgA IgE
IgM IgD

V D J C
 The heavy chain synthesis uses the same mechanisms of
rearrangement as that of L chain—the use of the V(D)J
recombinase to mediate the joining of different gene
segments.
 In the early stages of the life of a particular B cell, two
rearrangements of germ line DNA must occur. The 1st
brings one D segment alongside one J segment. The 2nd
brings one V segment to the DJ unit (fig.)
 The rearranged DNA is then transcribed along with the
closest C region genes  and .
Organization and rearrangement of heavy chain genes

IgG IgA IgE

IgM IgD

(On chromosome 14)

IgM IgD
Regulation of Ig Gene Expression
 This mechanism of allelic exlusion ensures that
every B cell and the antibodies it synthesizes are
monospecific.
 The arrangements occur in the absence of antigen in
the early stages of B cell differentiation.
 Class or isotype switching
• A single B cell can switch to make a different class of
antibody, such as IgG, IgE, or IgA. This phenomenon is
known as class or isotype switching. Class switching changes
the effector function of the B cell but does not change the
cell’s antigenic specificity.
• Class switching occurs in antigen-stimulated B cells
synthesizing IgM and IgD, and involves further DNA
rearrangement juxtaposing the rearranged VDJ genes with
different heavy chain.
• In addition to antigen, class switching is dependent on the
presence of cytokines released by T cells.
 The cytokine that affect class switching induce further
rearrangement of B cell DNA and produce switching to
other Ig classes in a downstream progression.
 The S region, at the end of every H chain C region,
permits any of the CH genes (other than C) to associate
with the VDJ unit (Fig.).
 The cytokines present when antigen activates B cells
play a key role in selection during isotope switching.
 Each cytokine is thought to loosen the DNA double
helix structure at only certain points along the Ig locus,
allowing an enzyme “ switch recombinase” to recognize
DNA coding for specific C region.
Class or isotype switching

(switch region)

IgG1
 Class switching allows an antibody with a single
antigenic specificity to associate with a variety of
different constant region chain thus have different
effector function.

For example. An antibody with a VDJ units specific

for a bacterial antigen may be linked to C to
produce an IgG molecules; IgG interact with cells
such as macrophages that express receptors for Fc.

The same VDJ unit may be linked to C to produce

an IgE molecule; IgE interact with cell such as mast
cells that express receptors for Fc.
Generation of antibody diversity
1. VJ and VDJ combinatorial association
The association of any V gene segment with any J
segment can occur to form L chain. The same, any V can
associate with any D or J segments to make H chain
L chains:
40 V X 5 J = 200  chains
40 V X 4 J = 160  chains
H Chains: 50 V X 20 D X 6 J = 6000 H chains

2. Random assortment of H and L chains

Any H chain may associate with any L chain, thus
1.2 x 106 different -containing Ig molecules (200 x6000)
and 0.96 x 106 different -containing Ig molecules (160
x6000)
 3. Junctional and Insertional Diversity
• The precise position at which the genes for the V ,
(D), and J segments are fused together are not
constant, and inprecise DNA recombination can lead
to changes in the amino acids at these junction sites
(junctional diversity) that affect the antigen-binding
site.
• In addition, small sets of nucleotides may be
inserted (insertional diversity, N region diversity) at
the V-D and D-J junctions (mediated by the enzyme
terminal deoxynucleotidytransferase ( TdT)
 4. Somatic hypermutaion
 Mutation that occur in V genes of heavy and/ or light chains during
the lifetime of a B cell also increase the variety of antibodies
produced.
 Generally an antibody of low affinity is produced in the primary
response to antigen, DNA and amino acid sequences of this
antibody closely match the sequences encoded by germ line DNA.
As the response matures, the affinity for antigen of the antibodies
synthesized increases, and the amino acid sequences of these
antibodies diverge form those coded for in the germ line DNA.
This divergence results predominantly from point mutaions in the
VDJ recombined unit of antibody V genes, which result in changes
in individual amino acids.
 This phenomenon is referred to as somatic hypermutaion because it
occurs at a rate at least 10000 fold higher that normal rate. This
results in the observed increased affinity of antibodies for antigen
in the secondary response.
 5. Somatic gene conversion
Many species other than humans and mice (e.g. birds and rabbits)
rely on somatic gene conversion (involving non-reciprocal
exchange of sequences among genes ) and somatic hypermutaion
to generate diversity within the primary Ig repertoire.
 6. Receptor editing
Under some circumstances, a cell in the B cell lineage can
undergo a second rearrangement of its L chain variable gene
segments, after it has formed a recombined VJ unit, this
process is known as receptor editing. Unrearranged
segments are used in the second rearrangement.

First rearrangement

Second rearrangement
 Brief:
1. VJ and VDJ combinatorial association
2. Random assortment of H and L chains
3. Junctional and insertional diversity
4. Somatic hypermutaion
5. Somatic gene conversion
6. Receptor editing
All these mechanisms contribute to the formation of a
huge library or repertoire of B lymphocytes that contain
all the specificities required to deal with the universe of
diverse epitopes. Estimates of the number of total Ig
specificities that can be generated in an individual are
on the order of 1015-1018
2. Blood groups, and cell-surface
molecules
 First transfusions unpredictable –some recovered and
some died.
 It was appreciated early in the 20th century that it was not
possible to transfuse blood at random between individuals.
 There are over 20 antigens (cell surface molecules) on
erythrocytes, and they vary between individuals.
 The most important and well known are those that
comprise the ABO blood group system.
 Karl Landsteiner described the human ABO blood groups
in 1901. He identified 3 types of blood (A, B, & O).
 In 1910, a rare 4thtype was found (AB)
 More than 26 blood types have been identified.
  ABO Blood Group
 Differences in glycosylation of red cell glycoproteins
give rise to three variants of the glycoprotein called A, B
or O. The groups are co-expressed so that individuals
can be O, A, B or AB.
 The O (or H) antigen is generated in all individuals, and
consists of a particular carbohydrate group that is added
to proteins.
 The ABO locus codes for a galactosyltransferase enzyme
that adds a further sugar group to the O antigen. The
specificity of this enzyme determines the blood group.
 A Allele—adds an extra N-acetylgalactosamine to O
antigen to form A antigen.
 B Allele—adds a galactose to O antigen to form B antigen.
 People with both transferases produce both A and B
antigen (AB blood type); those who lack these transferases
produce O antigen only (O blood type).
 An Example of Blood Type Incompatibility
 person with type A blood who is transfused with type B
blood will have antibodies that recognize and clump
the red blood cells carrying type B.
 Red blood cells burst, releasing hemoglobin.
 Rh Incompatibility
 The designation of blood type usually also includes
whether the person has or does not have the Rh factor on
the red blood cell. Rh individuals normally do not have
antibodies to the Rh factor, but they make them when
exposed to the Rh factor.
 Rh incompatibility occurs when occurs when an Rh- (no
Rh antigen)mother has an Rh+(has Rh antigen) child.
 If an Rh– mother has an Rh+ baby the woman will be
immunised against the Rh+ antigen during childbirth and
produce IgG antibodies to Rh.
 First Rh incompatible pregnancy—Fetal cells
recognized as foreign, Mother’s immune system attacks
fetal cells, Produces a mild reaction, few antibodies
present.
 Second Rh incompatible pregnancy—Large response,
plentiful antibodies, Destruction of fetal blood cells,
much damage.
 Other Blood Groups
 MNS system which also code for red-blood-cell
antigens. It is important in binding specific
glycoproteinson RBC

 Lewis encodes for enzyme (FUT3) that adds

antigen to fructose and the product of the H
gene necessary for ABO expression.

 Secretor gene causes A, B, H antigens in body

fluids.