Direct in vivo VH to JH rearrangement violating the 12/23 rule

V(D)J recombination at the immunoglobulin heavy chain (IgH) locus follows the 12/23 rule to ensure the correct assembly of the variable region gene segments. Here, we report characterization of an in vivo model that allowed us to study recombination violating the 12/23 rule, namely a mouse strain lacking canonical D elements in its IgH locus. We demonstrate that VH to JH joining can support the generation of all B cell subsets. However, the process is inefficient in that B cells and antibodies derived from the DH-less allele are not detectable if the latter is combined with a wild-type IgH allele. There is no preferential usage of any particular VH gene family or JH element in VHJH junctions, indicating that 23/23-guided recombination is possible, but is a low frequency event at the IgH locus in vivo.

[1]  Jian Ye.  The immunoglobulin IGHD gene locus in C57BL/6 mice , 2004, Immunogenetics.

[2]  D. Roth.  Restraining the V(D)J recombinase , 2003, Nature Reviews Immunology.

[3]  M. Gellert,et al.  Ordered assembly of the V(D)J synaptic complex ensures accurate recombination , 2002, The EMBO journal.

[4]  R. Jaenisch,et al.  Monoclonal mice generated by nuclear transfer from mature B and T donor cells , 2002, Nature.

[5]  K. Rajewsky,et al.  Homeostasis of Peripheral B Cells in the Absence of B Cell Influx from the Bone Marrow , 2001, The Journal of experimental medicine.

[6]  C. Schaniel,et al.  Selection events operating at various stages in B cell development. , 2001, Current opinion in immunology.

[7]  J. Kearney,et al.  B‐cell subsets and the mature ­preimmune repertoire. Marginal zone and B1 B cells as part of a “natural immune memory” , 2000, Immunological reviews.

[8]  M. Nussenzweig,et al.  V(D)J Recombination: Modulation of RAG1 and RAG2 Cleavage Activity on 12/23 Substrates by Whole Cell Extract and DNA-bending Proteins , 1997, The Journal of experimental medicine.

[9]  Marie-Paule Lefranc,et al.  IMGT, the international ImMunoGeneTics database® , 1997, Nucleic Acids Res..

[10]  K. Rajewsky,et al.  B cell development under the condition of allelic inclusion. , 1997, Immunity.

[11]  L. Herzenberg,et al.  An unbiased analysis of V(H)-D-J(H) sequences from B-1a, B-1b, and conventional B cells. , 1997, Journal of immunology.

[12]  Thomas B. Kepler,et al.  Interdependence of N Nucleotide Addition and Recombination Site Choice in V(D)J Rearrangement , 1996, The Journal of Immunology.

[13]  K. Rajewsky,et al.  Surrogate light chain expression is required to establish immunoglobulin heavy chain allelic exclusion during early B cell development. , 1996, Immunity.

[14]  S. Lukyanov,et al.  An improved PCR method for walking in uncloned genomic DNA. , 1995, Nucleic acids research.

[15]  K. Rajewsky,et al.  Analysis of the B-cell progenitor compartment at the level of single cells , 1994, Current Biology.

[16]  M. Lieber,et al.  Chimeric molecules created by gene amplification interfere with the analysis of somatic hypermutation of murine immunoglobulin genes. , 1994, Gene.

[17]  M. Schlissel,et al.  Double-strand signal sequence breaks in V(D)J recombination are blunt, 5'-phosphorylated, RAG-dependent, and cell cycle regulated. , 1993, Genes & development.

[18]  K. Rajewsky,et al.  Independent control of immunoglobulin switch recombination at individual switch regions evidenced through Cre-loxP-mediated gene targeting , 1993, Cell.

[19]  K. Rajewsky,et al.  A critical role of λ5 protein in B cell development , 1992, Cell.

[20]  E. Kabat,et al.  Identical V region amino acid sequences and segments of sequences in antibodies of different specificities. Relative contributions of VH and VL genes, minigenes, and complementarity-determining regions to binding of antibody-combining sites. , 1991, Journal of immunology.

[21]  M. Monk,et al.  HPRT-deficient (Lesch–Nyhan) mouse embryos derived from germline colonization by cultured cells , 1987, Nature.

[22]  S. Tonegawa.  Somatic generation of antibody diversity , 1983, Nature.

[23]  Marie-Paule Lefranc,et al.  IMGT, the international ImMunoGeneTics database , 1999, Nucleic Acids Res..

[24]  Marie-Paule Lefranc,et al.  IMGT, the international ImMunoGeneTics database , 1999, Nucleic Acids Res..

[25]  Marie-Paule Lefranc,et al.  IMGT, the international ImMunoGeneTics database , 2001, Nucleic Acids Res..

[26]  K. Rajewsky,et al.  A critical role of lambda 5 protein in B cell development. , 1992, Cell.

[27]  K. Rajewsky,et al.  Expansion and functional activity of Ly‐1+ B cells upon transfer of peritoneal cells into allotype‐congenic, newborn mice , 1987, European journal of immunology.

[28]  R. Hardy,et al.  Immunoglobulin‐bearing B cells reconstitute and maintain the murine Ly‐1 B cell lineage , 1986, European journal of immunology.