B cell and antibody repertoire development in rabbits: The requirement of gut-associated lymphoid tissues

doi:10.1016/j.dci.2005.06.017

Developmental & Comparative Immunology

Volume 30, Issues 1–2, 2006, Pages 137-153

https://doi.org/10.1016/j.dci.2005.06.017 Get rights and content

Abstract

The antibody repertoire of rabbits has interested immunologists for decades, in part because of the ease with which large quantities of high affinity antibodies can be obtained in serum, and in part because of the presence of genetic variants, allotypes, within V_H, C_H and C_L regions. Studies of these allotypes led to the initial descriptions of allelic exclusion, and neonatal suppression of serum Ig production (allotype suppression), and were instrumental in demonstrating that V and C regions are encoded by separate genes and are usually expressed in cis. The immune system of rabbit continues to be of interest primarily because of the use of both gene conversion and somatic hypermutation to diversify rearranged heavy and light chain genes and the role that gut-associated lymphoid tissues (GALT) and intestinal flora play in developing the primary (preimmune) antibody repertoire.

Section snippets

Rabbit germline repertoires

Germline genes encoding the constant regions of rabbit Ig heavy and light chains, as well as some allelic forms of C_H and C_L, are summarized in Table 1. V_H genes known to be utilized in productive VDJ gene rearrangements and heavy chain locus haplotypes are summarized in Table 2, Table 3, respectively. Four BAC clones that span approximately 0.5 Mb of the rabbit heavy chain locus have been sequenced using DNA from a donor of haplotype R1M F-I (Table 3; V_H1a2 allotype) [1]. Fig. 1 shows the maps

Rabbit immunoglobulin isotype expression

The approximate concentrations of rabbit isotypes in serum are given in Table 6. There is no evidence for IgD in the rabbit. Although early studies suggested there might be another surface immunoglobulin on IgM positive B cells that was not IgA or IgG [19], [20], [21], a gene encoding the δ heavy chain has not been identified in two independent cloning and sequencing studies of the region downstream of the Cμ gene [1], [2].

One of the most remarkable features of the rabbit IgH locus is the large

B lymphopoiesis

B lymphopoiesis begins in the fetal liver and omentum and switches, shortly before birth, to the bone marrow. B cell recombination excision circles (BRECs) can be found by 12 days of gestation in bone marrow, indicating that Ig gene rearrangements begin at this stage of development [28].

B lymphopoiesis proceeds by ordered rearrangement of IgH genes followed by IgL genes, as evidenced by the presence of IgH chains in progenitor B cells (preB cells) before IgL chains in B lymphopoietic tissues

Allotype suppression

The newborn rabbit is born with passively acquired maternal protective immunity. Using immunoglobulin allotypic markers, it has been possible to track the disappearance of the maternal immunoglobulins and to also detect the appearance of immunoglobulins synthesized by the young rabbit that could only have been inherited from their sires (reviewed in [38]). The appearance of expressed paternal allotype can be suppressed by exposure of developing fetal or neonatal rabbits to antibodies directed

Somatic diversification

Gene conversion of rearranged heavy and light chain sequences is found in rabbit appendix by 3–4 weeks of age. Somatic hypermutation occurred in the D and J regions, which lack known gene conversion donors [55], [56]. The region immediately 3′ of the J_H region is also diversified by somatic hypermutation and it is therefore likely that somatic hypermutation also occurs in the rearranged V_H gene segments [57], [58]. Small clones are found at 3–5 weeks; by 5.5 weeks, some larger clones are

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B cell and antibody repertoire development in rabbits: The requirement of gut-associated lymphoid tissues

Abstract

Section snippets

Rabbit germline repertoires

Rabbit immunoglobulin isotype expression

B lymphopoiesis

Allotype suppression

Somatic diversification

Gene

Gene

Cell

J Mol Biol

Cell

Immunity

Dev Comp Immunol

Cell Immunol

J Biol Chem

J Immunol Meth

J Immunl Meth

Mol Immunol

A new VH-CH recombinant in the rabbit

Immunogenetics

Molecular analysis of recombination sites within the immunoglobulin heavy chain locus of the rabbit

Immunogenetics

Altered phenotypic expression of immunoglobulin heavy-chain variable-region (VH) genes in Alicia rabbits probably reflects a small deletion in the VH genes closest to the joining region

Proc Natl Acad Sci USA

Mutation affecting the expression of immunoglobulin variable regions in the rabbit

Proc Natl Acad Sci USA

A second rabbit kappa isotype

J Exp Med

Multiplicity of constant kappa light chain genes in the rabbit genome: a b4b4 homozygous rabbit contains a k-bas gene

EMBO J

Mapping of the duplicated rabbit immunoglobulin kappa light chain locus

Eur J Immunol

Immunoglobulin lambda light chain evolution: Igl and Igl-like sequences form three major groups

Immunogenetics

Variant strain of rabbits lacking immunoglobulin k polypeptide chain

Nature

Lack of K1b9 light chains in Basilea rabbits is probably due to a mutation in an acceptor site for mRNA splicing

J Exp Med

Structural analysis of a rabbit immunoglobulin k2 J-C locus reveals multiple deletions

Nucleic Acids Res

Kappa-chain allotypes and isotypes in the rabbit: cDNA sequences of clones encoding b9 suggest an evolutionary pathway and possible role of the interdomain disulfide bond in quantitative allotype expression

Proc Natl Acad Sci USA

Evolution of genes for allelic and isotypic forms of immunoglobulin kappa chains and of the genes for T-cell receptor beta chains in rabbits

J Mol Evol

Generation of the primary antibody repertoire in rabbits: expression of a diverse set of Igk-V genes may compensate for limited combinatorial diversity at the heavy chain locus

Immunogenetics

Demonstration of a rabbit cell surface Ig that bears light chain and VH, but lacks μ-, α-, and γ-allotypes- rabbit IgD?

J Immunol

Expression of an unidentified immunoglobulin isotype on rabbit Ig-bearing lymphocytes

J Immunol

Identification of an IgD-like surface immunoglobulin on rabbit lymphocytes

Eur J Immunol

The IgA heavy-chain gene family in rabbit: cloning and sequence analysis of 13 Cα genes

EMBO J

Differential expression of 13 IgA-heavy chain genes in rabbit lymphoid tissues

J Immunol

Expression of 12 rabbit IgA Ca genes as chimeric rabbit-mouse IgA antibodies

Proc Natl Acad Sci USA

Functional analysis of I alpha promoter regions of multiple IgA heavy chain genes

J Immunol

Recombinant IgG molecules in rabbits doubly heterozygous for group a and group e allotypic specificities

Immunochemistry

Trans-chromosomal recombination within the Ig heavy chain switch region in B lymphocytes

Proc Natl Acad Sci USA

Fetal VDJ gene repertoire in rabbit: evidence for preferential rearrangement of VH1

Eur J Immunol

Pre-B and B cells in rabbits: ontogeny and allelic exclusion of Kappa light chain genes

J Exp Med

Immunofluorescence studies on the expression of VHa allotypes by pre-B and B cells of homozygous and heterozygous rabbits

Eur J Immunol

B lymphocyte development in rabbit: progenitor B cells and waning of B lymphopoiesis

J Immunol

Demonstration of a rabbit cell surface Ig that bears light chain and V_H, but lacks μ-, α-, and γ-allotypes- rabbit IgD?

Immunofluorescence studies on the expression of V_Ha allotypes by pre-B and B cells of homozygous and heterozygous rabbits