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

Somatic DNA rearrangements in B lymphocytes, including V(D)J gene rearrangements and isotype switching, generally occur in cis, i. e., intrachromosomally. We showed previously, however, that 3 to 7% of IgA heavy chains have the VH and Calpha regions encoded in trans. To determine whether the trans-association of VH and Calpha occurred by trans-chromosomal recombination, by trans-splicing, or by trans-chromosomal gene conversion, we generated and analyzed eight IgA-secreting rabbit hybridomas with trans-associated VH and Calpha heavy chains. By ELISA and by nucleotide sequence analysis we found that the VH and Calpha regions were encoded by genes that were in trans in the germline. We cloned the rearranged VDJ-Calpha gene from a fosmid library of one hybridoma and found that the expressed VH and Calpha genes were juxtaposed. Moreover, the juxtaposed VH and Calpha genes originated from different IgH alleles. From the same hybridoma, we also identified a fosmid clone with the other expected product of a trans-chromosomal recombination. The recombination breakpoint occurred within the Smicro/Salpha region, indicating that the trans-association of VH and Calpha genes occurred by trans-chromosomal recombination during isotype switching. We conclude that trans-chromosomal recombination occurs at an unexpectedly high frequency (7%) within the IgH locus of B lymphocytes in normal animals, which may explain the high incidence of B-cell tumors that arise from oncogene translocation into the IgH locus.

[1]  Heikyung Suh,et al.  Ku80 is required for immunoglobulin isotype switching , 1998, The EMBO journal.

[2]  R. Mage,et al.  VH mutant rabbits lacking the VH1a2 gene develop a2+ B cells in the appendix by gene conversion-like alteration of a rearranged VH4 gene. , 1998, Journal of immunology.

[3]  C. Tunyaplin,et al.  IgH gene rearrangements on the unexpressed allele in rabbit B cells. , 1997, Journal of immunology.

[4]  D. Ramsden,et al.  Specificity in V(D)J recombination: new lessons from biochemistry and genetics. , 1997, Current opinion in immunology.

[5]  P. L. Bergsagel,et al.  Promiscuous translocations into immunoglobulin heavy chain switch regions in multiple myeloma. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[6]  A. Saxon,et al.  Multiple types of chimeric germ-line Ig heavy chain transcripts in human B cells: evidence for trans-splicing of human Ig RNA. , 1996, Journal of immunology.

[7]  M. Kingzette,et al.  Evidence for limited B-lymphopoiesis in adult rabbits , 1996, The Journal of experimental medicine.

[8]  R. Chaganti,et al.  Chromosomal translocations cause deregulated BCL6 expression by promoter substitution in B cell lymphoma. , 1995, The EMBO journal.

[9]  K L Knight,et al.  Rabbit monoclonal antibodies: generating a fusion partner to produce rabbit-rabbit hybridomas. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[10]  P. Gearhart,et al.  Reciprocal homologous recombination in or near antibody VDJ genes , 1995, European journal of immunology.

[11]  M. Kingzette,et al.  Transchromosomally derived Ig heavy chains. , 1995, Journal of immunology.

[12]  H. Ohno,et al.  Class switch recombination of the immunoglobulin heavy chain gene frequently occurs in B-cell lymphomas associated with rearrangement of the BCL2 gene. , 1995, International journal of hematology.

[13]  T. Manser,et al.  Somatic generation of hybrid antibody H chain genes in transgenic mice via interchromosomal gene conversion , 1994, The Journal of experimental medicine.

[14]  T. McKeithan,et al.  Molecular characterization of the t(14;19)(q32;q13) translocation in chronic lymphocytic leukemia. , 1993, Leukemia.

[15]  M. Morrison,et al.  Cloning and sequencing of a t(14;19) breakpoint that involves the Cμ switch region , 1993, Genes, chromosomes & cancer.

[16]  T. Honjo,et al.  Synthesis and regulation of trans‐mRNA encoding the immunoglobulin ∊ heavy chain , 1993, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[17]  T. Manser,et al.  Somatic recombination of heavy chain variable region transgenes with the endogenous immunoglobulin heavy chain locus in mice. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[18]  B. Birren,et al.  Stable propagation of cosmid sized human DNA inserts in an F factor based vector. , 1992, Nucleic acids research.

[19]  R. T. D'Aquila,et al.  Maximizing sensitivity and specificity of PCR by pre-amplification heating , 1991, Nucleic Acids Res..

[20]  M. Nussenzweig,et al.  Trans-splicing as a possible molecular mechanism for the multiple isotype expression of the immunoglobulin gene , 1991, The Journal of experimental medicine.

[21]  J D Kemp,et al.  Resolution and characterization of pro-B and pre-pro-B cell stages in normal mouse bone marrow , 1991, The Journal of experimental medicine.

[22]  W. Frankel,et al.  Isotype switching of an immunoglobulin heavy chain transgene occurs by DNA recombination between different chromosomes , 1990, Cell.

[23]  W. Hanly,et al.  The IgA heavy‐chain gene family in rabbit: cloning and sequence analysis of 13 C alpha genes. , 1989, The EMBO journal.

[24]  R. D. Schneiderman,et al.  Expression of 12 rabbit IgA C alpha genes as chimeric rabbit-mouse IgA antibodies. , 1989, Proceedings of the National Academy of Sciences of the United States of America.

[25]  V. Oi,et al.  Transgenic rabbits with lymphocytic leukemia induced by the c-myc oncogene fused with the immunoglobulin heavy chain enhancer. , 1988, Proceedings of the National Academy of Sciences of the United States of America.

[26]  J. Uhr,et al.  Double isotype production by a neoplastic B cell line. II. Allelically excluded production of mu and gamma 1 heavy chains without CH gene rearrangement , 1986, The Journal of experimental medicine.

[27]  K. Rajewsky,et al.  Class switch recombination is IgG1 specific on active and inactive IgH loci of IgG1-secreting B-cell blasts. , 1986, Proceedings of the National Academy of Sciences of the United States of America.

[28]  T. Kipps,et al.  Homologous chromosome recombination generating immunoglobulin allotype and isotype switch variants. , 1986, The EMBO journal.

[29]  K. Knight,et al.  Organization and polymorphism of rabbit immunoglobulin heavy chain genes. , 1985, Journal of immunology.

[30]  P. Nowell,et al.  Cloning of the chromosome breakpoint of neoplastic B cells with the t(14;18) chromosome translocation. , 1984, Science.

[31]  E. Padlan,et al.  The structural and genetic basis for expression of normal and latent VHa allotypes of the rabbit. , 1984, Molecular immunology.

[32]  R. D. Schneiderman,et al.  Genes encoding α-heavy chains of rabbit IgA: characterization of cDNA encoding IgA-g subclass α-chains , 1984 .

[33]  P. Leder,et al.  Translocation of the c-myc gene into the immunoglobulin heavy chain locus in human Burkitt lymphoma and murine plasmacytoma cells. , 1982, Proceedings of the National Academy of Sciences of the United States of America.

[34]  M. Cole,et al.  Novel myc oncogene RNA from abortive immunoglobulin-gene recombination in mouse plasmacytomas , 1982, Cell.

[35]  T. Honjo,et al.  Nucleotide sequences of switch regions of immunoglobulin C epsilon and C gamma genes and their comparison. , 1982, The Journal of biological chemistry.

[36]  L. Stanton,et al.  On immunoglobulin heavy chain gene switching: two gamma 2b genes are rearranged via switch sequences in MPC-11 cells but only one is expressed. , 1982, Nucleic acids research.

[37]  Mark M. Davis,et al.  DNA sequences mediating class switching in alpha-immunoglobulins. , 1980, Science.

[38]  Hitoshi Sakano,et al.  Two types of somatic recombination are necessary for the generation of complete immunoglobulin heavy-chain genes , 1980, Nature.

[39]  J. Jaton,et al.  Independence of the antigen‐binding site and the VHa allotypic determinants of rabbit anti‐pneumococcal anti bodies , 1976 .

[40]  T. Malek,et al.  Recombinant rabbit secretory immunoglobulin molecules: alpha chains with maternal (paternal) variable-region allotypes and paternal (maternal) constant-region allotypes. , 1974, Proceedings of the National Academy of Sciences of the United States of America.

[41]  L. Forni,et al.  Heavy chain variable and constant region allotypes in single rabbit plasma cells. , 1973, Immunochemistry.

[42]  R. Mage,et al.  Distribution of allotypic specificities A1, A2, A14, and A15 among immunoglobulin G molecules. , 1970, Journal of immunology.

[43]  W. Hanly,et al.  Genetic control of alpha chains of rabbit IgA: allotypic specificities on the variable and the constant regions. , 1975, Contemporary topics in molecular immunology.