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. 1989 Jun 1;169(6):2007–2019. doi: 10.1084/jem.169.6.2007

Early onset of somatic mutation in immunoglobulin VH genes during the primary immune response

PMCID: PMC2189359  PMID: 2499654

Abstract

The dynamics of somatic mutation in Ig variable genes was investigated in order to define a population of B cells undergoing mutation. BALB/cJ mice were injected with PC-KLH, and splenic RNA was prepared 5, 7, and 13 d later. The mRNA was annealed to gamma constant region primers to make cDNA transcripts encoding VH genes. 103 cDNA clones corresponding to 18 different genes from the VH7183, VH3660, and VHS107 subfamilies were sequenced to identify mutation. VH genes had a low level of mutation on day 5 after immunization and accumulated more mutation by day 7 at a rate of 10(-3) mutations per nucleotide per generation. However, by day 13, the number of mutations per gene did not increase, and most of the substitutions encoded replacement amino acid changes that were clustered in the hypervariable regions, indicating that the mutational process was less active during the second week and that antigen selection had occurred. The data are consistent with a developmentally regulated mechanism in which mutation is activated during the first week of the primary immune response for a limited time period, after which selection acts to preserve the beneficial mutants.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Berek C., Milstein C. Mutation drift and repertoire shift in the maturation of the immune response. Immunol Rev. 1987 Apr;96:23–41. doi: 10.1111/j.1600-065x.1987.tb00507.x. [DOI] [PubMed] [Google Scholar]
  2. Brodeur P. H., Riblet R. The immunoglobulin heavy chain variable region (Igh-V) locus in the mouse. I. One hundred Igh-V genes comprise seven families of homologous genes. Eur J Immunol. 1984 Oct;14(10):922–930. doi: 10.1002/eji.1830141012. [DOI] [PubMed] [Google Scholar]
  3. Caton A. J., Brownlee G. G., Staudt L. M., Gerhard W. Structural and functional implications of a restricted antibody response to a defined antigenic region on the influenza virus hemagglutinin. EMBO J. 1986 Jul;5(7):1577–1587. doi: 10.1002/j.1460-2075.1986.tb04399.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Chirgwin J. M., Przybyla A. E., MacDonald R. J., Rutter W. J. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979 Nov 27;18(24):5294–5299. doi: 10.1021/bi00591a005. [DOI] [PubMed] [Google Scholar]
  5. Claflin J. L., Berry J., Flaherty D., Dunnick W. Somatic evolution of diversity among anti-phosphocholine antibodies induced with Proteus morganii. J Immunol. 1987 May 1;138(9):3060–3068. [PubMed] [Google Scholar]
  6. Claflin J. L., Berry J. Genetics of the phosphocholine-specific antibody response to Streptococcus pneumoniae. Germ-line but not mutated T15 antibodies are dominantly selected. J Immunol. 1988 Dec 1;141(11):4012–4019. [PubMed] [Google Scholar]
  7. Clarke S. H., Huppi K., Ruezinsky D., Staudt L., Gerhard W., Weigert M. Inter- and intraclonal diversity in the antibody response to influenza hemagglutinin. J Exp Med. 1985 Apr 1;161(4):687–704. doi: 10.1084/jem.161.4.687. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Coulondre C., Miller J. H., Farabaugh P. J., Gilbert W. Molecular basis of base substitution hotspots in Escherichia coli. Nature. 1978 Aug 24;274(5673):775–780. doi: 10.1038/274775a0. [DOI] [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. Cumano A., Rajewsky K. Structure of primary anti-(4-hydroxy-3-nitrophenyl)acetyl (NP) antibodies in normal and idiotypically suppressed C57BL/6 mice. Eur J Immunol. 1985 May;15(5):512–520. doi: 10.1002/eji.1830150517. [DOI] [PubMed] [Google Scholar]
  11. Dzierzak E. A., Janeway C. A., Jr, Richard N., Bothwell A. Molecular characterization of antibodies bearing Id-460. I. The structure of two highly homologous VH genes used to produce idiotype positive immunoglobulins. J Immunol. 1986 Mar 1;136(5):1864–1870. [PubMed] [Google Scholar]
  12. Feeney A. J., Clarke S. H., Mosier D. E. Specific H chain junctional diversity may be required for non-T15 antibodies to bind phosphorylcholine. J Immunol. 1988 Aug 15;141(4):1267–1272. [PubMed] [Google Scholar]
  13. 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]
  14. Gearhart P. J., Lawler A. M., Levy N. S., Lebecque S. G., Malipiero U. V. Generation of antibody diversity before and after immunization. Ann N Y Acad Sci. 1988;546:57–62. doi: 10.1111/j.1749-6632.1988.tb21619.x. [DOI] [PubMed] [Google Scholar]
  15. Golding G. B., Gearhart P. J., Glickman B. W. Patterns of somatic mutations in immunoglobulin variable genes. Genetics. 1987 Jan;115(1):169–176. doi: 10.1093/genetics/115.1.169. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Griffiths G. M., Berek C., Kaartinen M., Milstein C. Somatic mutation and the maturation of immune response to 2-phenyl oxazolone. Nature. 1984 Nov 15;312(5991):271–275. doi: 10.1038/312271a0. [DOI] [PubMed] [Google Scholar]
  17. 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]
  18. 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]
  19. Lawler A. M., Lin P. S., Gearhart P. J. Adult B-cell repertoire is biased toward two heavy-chain variable-region genes that rearrange frequently in fetal pre-B cells. Proc Natl Acad Sci U S A. 1987 Apr;84(8):2454–2458. doi: 10.1073/pnas.84.8.2454. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Li W. H., Wu C. I., Luo C. C. Nonrandomness of point mutation as reflected in nucleotide substitutions in pseudogenes and its evolutionary implications. J Mol Evol. 1984;21(1):58–71. doi: 10.1007/BF02100628. [DOI] [PubMed] [Google Scholar]
  21. Maizels N., Bothwell A. The T-cell-independent immune response to the hapten NP uses a large repertoire of heavy chain genes. Cell. 1985 Dec;43(3 Pt 2):715–720. doi: 10.1016/0092-8674(85)90244-2. [DOI] [PubMed] [Google Scholar]
  22. Near R. I., Juszczak E. C., Huang S. Y., Sicari S. A., Margolies M. N., Gefter M. L. Expression and rearrangement of homologous immunoglobulin VH genes in two mouse strains. Proc Natl Acad Sci U S A. 1984 Apr;81(7):2167–2171. doi: 10.1073/pnas.81.7.2167. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Ollo R., Sikorav J. L., Rougeon F. Structural relationships among mouse and human immunoglobulin VH genes in the subgroup III. Nucleic Acids Res. 1983 Nov 25;11(22):7887–7897. doi: 10.1093/nar/11.22.7887. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Riley S. C., Connors S. J., Klinman N. R., Ogata R. T. Preferential expression of variable region heavy chain gene segments by predominant 2,4-dinitrophenyl-specific BALB/c neonatal antibody clonotypes. Proc Natl Acad Sci U S A. 1986 Apr;83(8):2589–2593. doi: 10.1073/pnas.83.8.2589. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Sablitzky F., Wildner G., Rajewsky K. Somatic mutation and clonal expansion of B cells in an antigen-driven immune response. EMBO J. 1985 Feb;4(2):345–350. doi: 10.1002/j.1460-2075.1985.tb03635.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Shlomchik M. J., Marshak-Rothstein A., Wolfowicz C. B., Rothstein T. L., Weigert M. G. The role of clonal selection and somatic mutation in autoimmunity. 1987 Aug 27-Sep 2Nature. 328(6133):805–811. doi: 10.1038/328805a0. [DOI] [PubMed] [Google Scholar]
  27. Shlomchik M., Nemazee D., van Snick J., Weigert M. Variable region sequences of murine IgM anti-IgG monoclonal autoantibodies (rheumatoid factors). II. Comparison of hybridomas derived by lipopolysaccharide stimulation and secondary protein immunization. J Exp Med. 1987 Apr 1;165(4):970–987. doi: 10.1084/jem.165.4.970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Siekevitz M., Kocks C., Rajewsky K., Dildrop R. Analysis of somatic mutation and class switching in naive and memory B cells generating adoptive primary and secondary responses. Cell. 1987 Mar 13;48(5):757–770. doi: 10.1016/0092-8674(87)90073-0. [DOI] [PubMed] [Google Scholar]
  29. Winter E., Radbruch A., Krawinkel U. Members of novel VH gene families are found in VDJ regions of polyclonally activated B-lymphocytes. EMBO J. 1985 Nov;4(11):2861–2867. doi: 10.1002/j.1460-2075.1985.tb04015.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Wysocki L., Manser T., Gefter M. L. Somatic evolution of variable region structures during an immune response. Proc Natl Acad Sci U S A. 1986 Mar;83(6):1847–1851. doi: 10.1073/pnas.83.6.1847. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Yancopoulos G. D., Desiderio S. V., Paskind M., Kearney J. F., Baltimore D., Alt F. W. Preferential utilization of the most JH-proximal VH gene segments in pre-B-cell lines. Nature. 1984 Oct 25;311(5988):727–733. doi: 10.1038/311727a0. [DOI] [PubMed] [Google Scholar]

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