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. 1984 Jan 1;159(1):179–192. doi: 10.1084/jem.159.1.179

Multiple VH gene segments encode murine antistreptococcal antibodies

PMCID: PMC2187198  PMID: 6420495

Abstract

Most mouse strains are able to mount a diverse antibody response against group A streptococcal carbohydrate (GAC). We have previously reported that murine anti-GAC antibodies are for the most part restricted to IgM and IgG3 subclasses. In addition, despite extensive heterogeneity in their isoelectric focusing patterns, greater than 50% of A/J anti-GAC antibodies share a common light chain defined by spectrotypic and idiotypic (VK1GAC) criteria. We have used protein and DNA sequencing strategies to examine the genetic basis of diversity in murine anti-GAC antibodies. In particular, we report that, (a) multiple, closely homologous VH gene segments contribute to the generation of anti-GAC antibodies, (b) a common framework sequence, related to the VK27 subgroup, probably defines VK1GAC, and (c) the A/J anti-GAC VH regions and BALB/c anti-inulin VH sequences are 95% homologous at the protein level and are likely encoded by overlapping VH gene families. Lastly, we discuss the genetic mechanisms that might permit the evolution of multiple, closely homologous germline VH gene segments in the context of highly divergent flanking region sequences.

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Selected References

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  1. Anderson S., Gait M. J., Mayol L., Young I. G. A short primer for sequencing DNA cloned in the single-stranded phage vector M13mp2. Nucleic Acids Res. 1980 Apr 25;8(8):1731–1743. doi: 10.1093/nar/8.8.1731. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Baltimore D. Gene conversion: some implications for immunoglobulin genes. Cell. 1981 Jun;24(3):592–594. doi: 10.1016/0092-8674(81)90082-9. [DOI] [PubMed] [Google Scholar]
  3. Blin N., Stafford D. W. A general method for isolation of high molecular weight DNA from eukaryotes. Nucleic Acids Res. 1976 Sep;3(9):2303–2308. doi: 10.1093/nar/3.9.2303. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bothwell A. L., Paskind M., Reth M., Imanishi-Kari T., Rajewsky K., Baltimore D. Heavy chain variable region contribution to the NPb family of antibodies: somatic mutation evident in a gamma 2a variable region. Cell. 1981 Jun;24(3):625–637. doi: 10.1016/0092-8674(81)90089-1. [DOI] [PubMed] [Google Scholar]
  5. Briles D. E., Carroll R. J. A simple method for estimating the probable numbers of different antibodies by examining the repeat frequencies of sequences or isoelectric focusing patterns. Mol Immunol. 1981 Jan;18(1):29–38. doi: 10.1016/0161-5890(81)90045-6. [DOI] [PubMed] [Google Scholar]
  6. Briles D. E., Krause R. M. Mouse strain-specific idiotypy and interstrain idiotypic cross-reactions. J Immunol. 1974 Aug;113(2):522–530. [PubMed] [Google Scholar]
  7. Capra J. D., Berek C., Eichmann K. Structural studies on induced antibodies with defined idiotypic specificities. III. N-terminal amino acid sequence of the heavy and light chains of mouse anti-streptococcal antibodies--A5A, S8, and S117. J Immunol. 1976 Jul;117(1):7–10. [PubMed] [Google Scholar]
  8. Coligan J. E., Schnute W. C., Jr, Kindt T. J. Immunochemical and chemical studies on streptococcal group-specific carbohydrates. J Immunol. 1975 Jun;114(6):1654–1658. [PubMed] [Google Scholar]
  9. Crews S., Griffin J., Huang H., Calame K., Hood L. A single VH gene segment encodes the immune response to phosphorylcholine: somatic mutation is correlated with the class of the antibody. Cell. 1981 Jul;25(1):59–66. doi: 10.1016/0092-8674(81)90231-2. [DOI] [PubMed] [Google Scholar]
  10. Denhardt D. T. A membrane-filter technique for the detection of complementary DNA. Biochem Biophys Res Commun. 1966 Jun 13;23(5):641–646. doi: 10.1016/0006-291x(66)90447-5. [DOI] [PubMed] [Google Scholar]
  11. Eichmann K. Idiotype expression and the inheritance of mouse antibody clones. J Exp Med. 1973 Mar 1;137(3):603–621. doi: 10.1084/jem.137.3.603. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Gearhart P. J., Bogenhagen D. F. Clusters of point mutations are found exclusively around rearranged antibody variable genes. Proc Natl Acad Sci U S A. 1983 Jun;80(11):3439–3443. doi: 10.1073/pnas.80.11.3439. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Gearhart P. J., Johnson N. D., Douglas R., Hood L. IgG antibodies to phosphorylcholine exhibit more diversity than their IgM counterparts. Nature. 1981 May 7;291(5810):29–34. doi: 10.1038/291029a0. [DOI] [PubMed] [Google Scholar]
  14. Gershenfeld H. K., Tsukamoto A., Weissman I. L., Joho R. Somatic diversification is required to generate the V kappa genes of MOPC 511 and MOPC 167 myeloma proteins. Proc Natl Acad Sci U S A. 1981 Dec;78(12):7674–7678. doi: 10.1073/pnas.78.12.7674. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Herbst H., Chang J. Y., Aebersold R., Braun D. G. Murine VK25 isotype sequence: monoclonal antibody 2S1.3 specific for the group A streptococcal polysaccharide. Hoppe Seylers Z Physiol Chem. 1982 Sep;363(9):1069–1076. doi: 10.1515/bchm2.1982.363.2.1069. [DOI] [PubMed] [Google Scholar]
  16. Hohn B., Murray K. Packaging recombinant DNA molecules into bacteriophage particles in vitro. Proc Natl Acad Sci U S A. 1977 Aug;74(8):3259–3263. doi: 10.1073/pnas.74.8.3259. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Honjo T. Immunoglobulin genes. Annu Rev Immunol. 1983;1:499–528. doi: 10.1146/annurev.iy.01.040183.002435. [DOI] [PubMed] [Google Scholar]
  18. Hood L., Campbell J. H., Elgin S. C. The organization, expression, and evolution of antibody genes and other multigene families. Annu Rev Genet. 1975;9:305–353. doi: 10.1146/annurev.ge.09.120175.001513. [DOI] [PubMed] [Google Scholar]
  19. Hunkapiller M. W., Hood L. E. Direct microsequence analysis of polypeptides using an improved sequenator, a nonprotein carrier (polybrene), and high pressure liquid chromatography. Biochemistry. 1978 May 30;17(11):2124–2133. doi: 10.1021/bi00604a016. [DOI] [PubMed] [Google Scholar]
  20. Hunkapiller M. W., Hood L. E. New protein sequenator with increased sensitivity. Science. 1980 Feb 1;207(4430):523–525. doi: 10.1126/science.7352258. [DOI] [PubMed] [Google Scholar]
  21. Kaartinen M., Griffiths G. M., Markham A. F., Milstein C. mRNA sequences define an unusually restricted IgG response to 2-phenyloxazolone and its early diversification. 1983 Jul 28-Aug 3Nature. 304(5924):320–324. doi: 10.1038/304320a0. [DOI] [PubMed] [Google Scholar]
  22. Kim S., Davis M., Sinn E., Patten P., Hood L. Antibody diversity: somatic hypermutation of rearranged VH genes. Cell. 1981 Dec;27(3 Pt 2):573–581. doi: 10.1016/0092-8674(81)90399-8. [DOI] [PubMed] [Google Scholar]
  23. Kraig E., Kronenberg M., Kapp J. A., Pierce C. W., Abruzzini A. F., Sorensen C. M., Samelson L. E., Schwartz R. H., Hood L. E. T and B cells that recognize the same antigen do not transcribe similar heavy chain variable region gene segments. J Exp Med. 1983 Jul 1;158(1):192–209. doi: 10.1084/jem.158.1.192. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Kurosawa Y., Tonegawa S. Organization, structure, and assembly of immunoglobulin heavy chain diversity DNA segments. J Exp Med. 1982 Jan 1;155(1):201–218. doi: 10.1084/jem.155.1.201. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Leder P. Genetic control of immunoglobulin production. Hosp Pract (Off Ed) 1983 Feb;18(2):73–82. doi: 10.1080/21548331.1983.11702475. [DOI] [PubMed] [Google Scholar]
  26. Loenen W. A., Brammar W. J. A bacteriophage lambda vector for cloning large DNA fragments made with several restriction enzymes. Gene. 1980 Aug;10(3):249–259. doi: 10.1016/0378-1119(80)90054-2. [DOI] [PubMed] [Google Scholar]
  27. Maniatis T., Hardison R. C., Lacy E., Lauer J., O'Connell C., Quon D., Sim G. K., Efstratiadis A. The isolation of structural genes from libraries of eucaryotic DNA. Cell. 1978 Oct;15(2):687–701. doi: 10.1016/0092-8674(78)90036-3. [DOI] [PubMed] [Google Scholar]
  28. Mullins J. I., Casey J. W., Nicolson M. O., Davidson N. Sequence organization of feline leukemia virus DNA in infected cells. Nucleic Acids Res. 1980 Aug 11;8(15):3287–3305. doi: 10.1093/nar/8.15.3287. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Nahm M. H., Clevinger B. L., Davie J. M. Monoclonal antibodies to streptococcal group A carbohydrate. I. A dominant idiotypic determinant is located on Vk. J Immunol. 1982 Oct;129(4):1513–1518. [PubMed] [Google Scholar]
  30. Perlmutter R. M., Briles D. E., Davie J. M. Complete sharing of light chain spectrotypes by murine IgM and IgG anti-streptococcal antibodies. J Immunol. 1977 Jun;118(6):2161–2166. [PubMed] [Google Scholar]
  31. Perlmutter R. M., Briles D. E., Greve J. M., Davie J. M. Light chain diversity of murine anti-streptococcal antibodies: IgCH-linked effects on L chain expression. J Immunol. 1978 Jul;121(1):149–158. [PubMed] [Google Scholar]
  32. Perlmutter R. M., Hansburg D., Briles D. E., Nicolotti R. A., Davie J. M. Subclass restriction of murine anti-carbohydrate antibodies. J Immunol. 1978 Aug;121(2):566–572. [PubMed] [Google Scholar]
  33. Potter M., Newell J. B., Rudikoff S., Haber E. Classification of mouse VK groups based on the partial amino acid sequence to the first invariant tryptophan: impact of 14 new sequences from IgG myeloma proteins. Mol Immunol. 1982 Dec;19(12):1619–1630. doi: 10.1016/0161-5890(82)90273-5. [DOI] [PubMed] [Google Scholar]
  34. Rigby P. W., Dieckmann M., Rhodes C., Berg P. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol. 1977 Jun 15;113(1):237–251. doi: 10.1016/0022-2836(77)90052-3. [DOI] [PubMed] [Google Scholar]
  35. Rocca-Serra J., Matthes H. W., Kaartinen M., Milstein C., Thèze J., Fougereau M. Analysis of antibody diversity: V-D-J mRNA nucleotide sequence of four anti-GAT monoclonal antibodies. A paucigene system using alternate D-J recombinations to generate functionally similar hypervariable regions. EMBO J. 1983;2(6):867–872. doi: 10.1002/j.1460-2075.1983.tb01515.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Sakano H., Maki R., Kurosawa Y., Roeder W., Tonegawa S. Two types of somatic recombination are necessary for the generation of complete immunoglobulin heavy-chain genes. Nature. 1980 Aug 14;286(5774):676–683. doi: 10.1038/286676a0. [DOI] [PubMed] [Google Scholar]
  37. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Selsing E., Storb U. Somatic mutation of immunoglobulin light-chain variable-region genes. Cell. 1981 Jul;25(1):47–58. doi: 10.1016/0092-8674(81)90230-0. [DOI] [PubMed] [Google Scholar]
  39. Siekevitz M., Huang S. Y., Gefter M. L. The genetic basis of antibody production: a single heavy chain variable region gene encodes all molecules bearing the dominant anti-arsonate idiotype in the strain A mouse. Eur J Immunol. 1983 Feb;13(2):123–132. doi: 10.1002/eji.1830130207. [DOI] [PubMed] [Google Scholar]
  40. Smith G. E., Summers M. D. The bidirectional transfer of DNA and RNA to nitrocellulose or diazobenzyloxymethyl-paper. Anal Biochem. 1980 Nov 15;109(1):123–129. doi: 10.1016/0003-2697(80)90019-6. [DOI] [PubMed] [Google Scholar]
  41. Tonegawa S. Somatic generation of antibody diversity. Nature. 1983 Apr 14;302(5909):575–581. doi: 10.1038/302575a0. [DOI] [PubMed] [Google Scholar]
  42. Vrana M., Rudikoff S., Potter M. Sequence variation among heavy chains from inulin-binding myeloma proteins. Proc Natl Acad Sci U S A. 1978 Apr;75(4):1957–1961. doi: 10.1073/pnas.75.4.1957. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Zimmerman C. L., Appella E., Pisano J. J. Rapid analysis of amino acid phenylthiohydantoins by high-performance liquid chromatography. Anal Biochem. 1977 Feb;77(2):569–573. doi: 10.1016/0003-2697(77)90276-7. [DOI] [PubMed] [Google Scholar]

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