Immunoglobulin Genetics

Immunoglobulin Genetics
History
Same C region could associate with many V regions
IgG Ab with different specificities

Same V region could associate with many C regions
Same idiotype on IgG and IgM Ab

V and C regions coded for by separate genes

Light Chain Gene Families
Germ line gene organization
J

1
C

1
E
J

2
C

2
E
J

3
C

3
E
J

4
C

4
E P
L V

1
P
L V

n
P
L V

2
Lambda light chain genes; n=30
P
L V

1
P
L V

n
P
L V

2 J

2 J

3 J

5 C

E
J

1 J

4
Kappa light chain genes; n=300
Light Chain Gene Families
Gene rearrangement and expression
L V

1 L V

n L V

2 J

2 J

3 J

5 C

1 J

4
DNA
P P P E
P P E
L V

1 L J

5 C

2 J

4
DNA
DNA Rearrangement
V C J
V C
Protein
Transport to ER
Protein
Translation
C L V J
E
L V C

J
RNA
Transcription
Primary transcript
C L V J
RNA Processing
RNA
mRNA
Heavy Chain Gene Family
Germ line gene organization
Introns separate exons coding for H chain domains
Heavy chain genes; Vn=1000, Dn=15
P
L V1
P
L Vn
P
L V2 D2 D3 D1 Dn
C

3

C

C

2
C

1
C

4
C

1

C

2

J2 J3 J5 J1 J4
E
C
H
1
H
C
H
2 C
H
3 C
H
4
Heavy Chain Gene Family
Gene rearrangement and expression
P
L V1
P
L Vn
P
L V2 D2 D3 D1 Dn
C

J2 J3 J5 J1 J4
E
DJ rearrangement
P
L Vn
P
L V1
P
L V2
D2
D1
C

J5
J4
E
DNA
VDJ rearrangement
D2
C

J5
J4
E P
L V1
P
L
V2
DNA
Transcription
D2
C

J5
J4
E
L
V2
RNA
Primary transcript
Heavy Chain Gene Family
Gene rearrangement and expression
E
Primary transcript
E
J4 D2 V2
C

J5 L
C

J5 L
D2 J4 V2
RNA Processing
A
n

mRNA for C

L D J V
C

L D J V
A
n

mRNA for C

Translation
C

heavy chain
C

L D J V C

L D J V
C

heavy
chain
Transport to ER
C

heavy chain
V C
C

D J V C

D J V
C

heavy
chain
V C
Mechanism of DNA Rearrangements
Recombination
signal sequences
(RSS)
Nonmer
Heptamer
1 or 2 turn signals
Rag-1 and Rag-2
L V
2
J C
1
Lambda light chains
L V
1
J C
2
Kappa light chains
Heavy chains
L V
2
J C
2 1 1
D
Heptamer
CACAGTG
GTGTCAC
Nonamer
ACAAAAACC
TGTTTTTGG
23 bp
23 bp
Two turn
RSS
2
Nonamer
GGTTTTTGT
CCAAAAACA
Heptamer
CACTGTG
GTGACAC
12 bp
12 bp
One turn
RSS
1
Order of Ig Gene Expression
+
-
Heavy chain gene
VDJ recombination
Productive rearrangement ?
Yes
No
Transcribe/translate
1st
2nd
or
Apoptosis
Progenitor B cell
Pre B cell

() Heavy chain
Paternal Maternal
Order of Ig Gene Expression
+
-
Light chain gene
VJ recombination
Productive rearrangement ?
Yes
No
Transcribe/translate
Light chain
4 th
Apoptosis
1st -
2nd -
3rd -
4th -

or
Pre B cell
Mature B cell


+

Order of Ig Gene Expression
Explains:
Why an individual B cell only produces one
kind of Ig with one kind of heavy and one
kind of light chain
Why an individual B cell makes only makes
antibodies of one specificity
Why there is allelic exclusion
Origin of Antibody Diversity
History
Definition Diversity (repitoire) is the total
of all the Ab specificities that an organism
is capable of expressing
Theories
Germ line theory
Somatic mutation theory
Origin of Antibody Diversity
Current concepts
Multiple V genes
V-J and V-D-J joining
Junctional diversity
N region insertions
Amino acid
sequences not
encoded in the germ
line

D region
T G G C C C
Pro Trp
C C C C C C
V gene
T G G
D region
C C C C C C
V gene
T G G
C G G C C C
Pro Arg
C C C C C C
V gene
T G G
D region
C G C C C C
Pro Pro
-GTCAATG
-GTCAATG- +
Break DNA
TdT adds nucleotides
Ligate
N region insertion Intron
J D
J
J
J
D
D
D
Generation of N region insertions
Origin of Antibody Diversity
Current concepts
Multiple V genes
V-J and V-D-J joining
Junctional diversity
N region insertions
Somatic mutation
Combinatorial association
Multispecificity
Generation of Diversity
B cell receptor
(Immunoglobulin)
Heavy Light
V gene segments 1000 300
D gene segments 15 -
J gene segments 4 4
N region insertion ++ -
Junctional diversity +++ +
Somatic mutation + +

V x D x J
1000 x 15 x 4
V x J
300 x 4
Total 6 x 10
4
1.2 x 10
3

Combinatorial association 7.2 x 10
7



Ag independent process Clonal selection
T Cell Antigen Receptor
Heterodimer of an and a chain
Gene structure and expression of the chain is
analogous to the immunoglobulin heavy chain
Gene structure and expression of the chain is
analogous to the immunoglobulin light chain
No somatic mutation in T cells