7Genetics of Immunoglobulins
Janardan P. Pandey
Department of Microbiology and Immunology, Medical University of South Carolina,
Charleston, South Carolina, U.S.A.
INTRODUCTION
Human immunoglobulin (Ig) molecules are coded by three unlinked gene families: two for light (L) chains
located on chromosomes 2 (k chains) and 22 (l chains), and one for heavy (H) chains located on
chromosome 14. As mentioned in the preceding chapters, each individual is able to produce billions of
antibody molecules with different antigenic specificities, and this diversity corresponds to the extreme
heterogeneity of the variable (V) regions in those antibody molecules, implying that each individual must
possess a large number of structural genes for Ig chains. The allotypic determinants on the constant (C)
region (see following discussion), on the other hand, segregate as a single Mendelian trait, suggesting that
there may be only one gene for each of the several Ig chain C regions. To reconcile these seemingly
contradictory observations, Dreyer and Bennet, in 1965, proposed that two separate genes that are brought
together by a translocation event during lymphocyte development encode the V and C regions. Employing
recombinant DNA technology, Hozumi and Tonegawa, in 1976, obtained conclusive proof of this
hypothesis (for his seminal studies, Tonegawa was awarded the 1987 Nobel prize in Medicine and
Physiology).
IMMUNOGLOBULIN GENE REARRANGEMENT
It is well-established that an Ig polypeptide chain is coded by multiple genes scattered along a
chromosome of the germ-line genome. These widely separated gene segments are brought together
(recombined) during B-lymphocyte differentiation to form a complete Ig gene. The V regions of the Ig-L
chains are coded by two gene segments, designated as V and J (J for joining, because it joins V and C
region genes). Three gene segments are required for the synthesis of the V region of the H chains: V, J,
and D (D for diversity, corresponds to the most diverse region of the H chain). To form a functional L or
H chain gene, one or two gene rearrangements are needed. On chromosomes 2 or 22, a V gene moves
next to a J gene. On chromosome 14, first the D and J regions are joined, and next one of the V genes is
joined to the DJ complex. The VJ segments and one of the L chain C regions (Ckor Cl) or VDJ segments
and one of the CH-gene complexes (Cm, Cd, Cg3, Cg1, Ca1, Cg2, Cg4, C1, or Ca2) are then transcribed
into nuclear RNA that contains these sequences as well as the interconnecting noncoding sequences. The
intervening noncoding sequences are then excised making a contiguous VJC mRNA for an L chain and a
contiguous VDJC mRNA for an H chain (Figs. 1 and 2). Gene rearrangements occur in a sequential order:
usually H chain genes rearrange first, followed by kchain genes and lastly by lgenes.
It has been shown that the VDJ joining is regulated by two proteins encoded by two closely linked
recombination-activating genes (RAG-1 and RAG-2) localized on the short arm of human chromosome
11. These genes have at least two unusual characteristics not shared by most eukaryotic genes: they are
devoid of introns and, although adjacent in location and synergistic in function, they have no sequence
homology. The latter implies that, unlike the Ig and major histocompatibility complex (MHC) genes,
RAG-1 and RAG-2 did not arise by gene duplication. Recent studies suggest that these genes may be
evolutionarily related to transposons, genetic elements that can be transposed in the genome from one
location to the other. Conserved recombination signal sequences (RSS) serve as substrate for the enzymes
coded by the RAG genes. These enzymes introduce a break between the RSS and the
74 Pandey
FIGURE 1 The embryonic DNA of chromosome 2 contains over 300 variable (V) genes, five joining (J) genes, and a
constant (C) gene (A). The V and J gene code for the kappa chain’s V region, C for its C region. In the left pathway,
differentiation of the embryonic cell to a plasma cell results in deletion of the intervening V genes so that V k1 is joined
with the J1 gene (B). The linked Vk1J1 segment codes for one of over 1,500 possible kappa light chain V regions. The
plasma cell DNA is transcribed into nuclear RNA (C). Splicing of the nuclear RNA produces messenger RNAs with the
Vk1, J1, and C genes linked together (D), ready for translation of a kappa light chain protein (E). The alternate pathway
at right (B–D) shows another of the many possible pathways leading to a different kappa light chain with a different
variable region specificity. Source: Modified from David JR. Antibodies: structure and function. Scientific American
Medicine. New York: Scientific American Inc., 1980.
FIGURE 2 A stretch of embryonic DNA in chromosome 14 contains a section coding for the heavy-chain variable
region; this DNA is made up of at least 100 variable (V) genes, 50 diversity (D) genes, and four to six joining (J) genes.
The section coding for the constant (C) region is formed by nine C genes (A). In the pathway shown, when the
embryonic cell differentiates into a plasma cell, some V and D genes are deleted so that V 1, D4, and J1 are joined to
form one of many possible heavy-chain genes (B). The plasma cell DNA is then transcribed into nuclear RNA (C). RNA
splicing selects the C gene and joins it to the V1, D4, and J1 genes (D). The resulting messenger RNAs will code for IgM
heavy chains (E). If RNA splicing removes the Cmm piece from the Cm gene, the IgM will be secreted. If the piece
remains, the IgM will be membrane-bound. Source: Modified from David JR. Antibodies: structure and function.
Scientific American Medicine. New York: Scientific American Inc., 1980.
Genetics of Immunoglobulins 75
FIGURE 3 In the secondary response, a plasma cell switches from IgM production (A) to IgG2 production by deleting
a DNA loop containing the constant (C)-region genes Cm, C@, Cg3, Cg1, and Cm1 from the IgM heavy (H)-chain gene
(B). This DNA is now transcribed into nuclear RNA (C). RNA splicing links the Cg2 gene with the J1 gene (D), and then
the mRNA is translated into an IgG2 heavy chain (E). Source: Modified from David JR. Antibodies: structure and
function. Scientific American Medicine. New York: Scientific American Inc., 1980.
coding sequence. Mechanisms involved in subsequent rejoining to form a mature coding segment are not
completely understood.
The transcription of Ig genes, like other eukaryotic genes, is regulated by promoters and enhancers.
Promoters, located 50 of the V segment, are necessary for transcription initiation. Enhancers, located in
the introns between J and C segments, increase the rate of transcription. For this reason, Ig synthesis (H
or L chains) is only detected after the VDJ or VJ rearrangements, which bring the promoter in close
proximity to the enhancer.
During ontogeny and functional differentiation, the H chain genes may undergo further gene
rearrangements that result in Ig class switching. As the B-lymphocytes differentiate into plasma cells, one
H chain C gene segment can be substituted for another without alteration of the VDJ combination (Fig.
3). In other words, a given V region gene can be expressed in association with more than one H chain
class or subclass, so that at the cellular level the same antibody specificity can be associated with the
synthesis of an IgM Ig (characteristic of the early stages of ontogeny and of the primary response) or with
an IgG Ig (characteristic of the mature individual and of the secondary response). Ig class switching is the
result of an intrachromosomal recombination between the switch region of Cm and one of the downstream
switch regions. This recombination event leads to looping out deletion of the intervening DNA segment
and joins the rearranged V region to a different C region (Cg, Ce, and Ca).