Analysis of somatic hypermutation in mouse Peyer's patches using immunoglobulin kappa light-chain transgenes (original) (raw)

Abstract

We have exploited mice transgenic for an immunoglobulin kappa light chain in order to show that immunoglobulin genes in the B cells of Peyer's patches in unimmunized mice carry a high level of somatic mutations. Most of the mutations are found in the subpopulation of B cells which, based on peanut agglutinin binding, derive from the germinal centers. The number of mutations per clone and their distribution along the variable gene segment (indicative of untemplated point mutations) are very similar to those found in antigen-specific splenic B cells of normal mice after secondary immunization. The mutations accumulate mainly in complementarity-determining region 1, in particular in some specific codons (Ser-26, Ser-31, and Ser-77) which have been previously recognized as intrinsic hypermutational hotspots. These results suggest that, as in the spleen, somatic mutation occurs in B cells which have migrated to the germinal centers, probably as a consequence of stimulation by antigens present in the gut environment. Transgenic animals are increasingly being used to define the signals involved in hypermutation. However, their subsequent study is very time-consuming because it is based on immunization and analysis of hybridomas or antigen-selected cells. We propose that the use of Peyer's patches of unimmunized adult mice offers a reliable and simple approach to analyze hypermutation of transgenes.

9862

Selected References

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

  1. Berek C., Berger A., Apel M. Maturation of the immune response in germinal centers. Cell. 1991 Dec 20;67(6):1121–1129. doi: 10.1016/0092-8674(91)90289-b. [DOI] [PubMed] [Google Scholar]
  2. Berek C., Griffiths G. M., Milstein C. Molecular events during maturation of the immune response to oxazolone. Nature. 1985 Aug 1;316(6027):412–418. doi: 10.1038/316412a0. [DOI] [PubMed] [Google Scholar]
  3. Betz A. G., Neuberger M. S., Milstein C. Discriminating intrinsic and antigen-selected mutational hotspots in immunoglobulin V genes. Immunol Today. 1993 Aug;14(8):405–411. doi: 10.1016/0167-5699(93)90144-a. [DOI] [PubMed] [Google Scholar]
  4. Betz A. G., Rada C., Pannell R., Milstein C., Neuberger M. S. Passenger transgenes reveal intrinsic specificity of the antibody hypermutation mechanism: clustering, polarity, and specific hot spots. Proc Natl Acad Sci U S A. 1993 Mar 15;90(6):2385–2388. doi: 10.1073/pnas.90.6.2385. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Butcher E. C., Rouse R. V., Coffman R. L., Nottenburg C. N., Hardy R. R., Weissman I. L. Surface phenotype of Peyer's patch germinal center cells: implications for the role of germinal centers in B cell differentiation. J Immunol. 1982 Dec;129(6):2698–2707. [PubMed] [Google Scholar]
  6. Coffman R. L., Weissman I. L. B220: a B cell-specific member of th T200 glycoprotein family. Nature. 1981 Feb 19;289(5799):681–683. doi: 10.1038/289681a0. [DOI] [PubMed] [Google Scholar]
  7. Even J., Griffiths G. M., Berek C., Milstein C. Light chain germ-line genes and the immune response to 2-phenyloxazolone. EMBO J. 1985 Dec 16;4(13A):3439–3445. doi: 10.1002/j.1460-2075.1985.tb04102.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Giusti A. M., Manser T. Hypermutation is observed only in antibody H chain V region transgenes that have recombined with endogenous immunoglobulin H DNA: implications for the location of cis-acting elements required for somatic mutation. J Exp Med. 1993 Mar 1;177(3):797–809. doi: 10.1084/jem.177.3.797. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. 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]
  10. Hermans M. H., Wubbena A., Kroese F. G., Hunt S. V., Cowan R., Opstelten D. The extent of clonal structure in different lymphoid organs. J Exp Med. 1992 May 1;175(5):1255–1269. doi: 10.1084/jem.175.5.1255. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Jacob J., Kelsoe G., Rajewsky K., Weiss U. Intraclonal generation of antibody mutants in germinal centres. Nature. 1991 Dec 5;354(6352):389–392. doi: 10.1038/354389a0. [DOI] [PubMed] [Google Scholar]
  12. Keohavong P., Thilly W. G. Fidelity of DNA polymerases in DNA amplification. Proc Natl Acad Sci U S A. 1989 Dec;86(23):9253–9257. doi: 10.1073/pnas.86.23.9253. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Kroese F. G., Wubbena A. S., Seijen H. G., Nieuwenhuis P. Germinal centers develop oligoclonally. Eur J Immunol. 1987 Jul;17(7):1069–1072. doi: 10.1002/eji.1830170726. [DOI] [PubMed] [Google Scholar]
  14. Lozano F., Rada C., Jarvis J. M., Milstein C. Affinity maturation leads to differential expression of multiple copies of a kappa light-chain transgene. Nature. 1993 May 20;363(6426):271–273. doi: 10.1038/363271a0. [DOI] [PubMed] [Google Scholar]
  15. MacLennan I. C., Liu Y. J., Oldfield S., Zhang J., Lane P. J. The evolution of B-cell clones. Curr Top Microbiol Immunol. 1990;159:37–63. doi: 10.1007/978-3-642-75244-5_3. [DOI] [PubMed] [Google Scholar]
  16. Meyer K. B., Sharpe M. J., Surani M. A., Neuberger M. S. The importance of the 3'-enhancer region in immunoglobulin kappa gene expression. Nucleic Acids Res. 1990 Oct 11;18(19):5609–5615. doi: 10.1093/nar/18.19.5609. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Milstein C., Even J., Jarvis J. M., Gonzalez-Fernandez A., Gherardi E. Non-random features of the repertoire expressed by the members of one V kappa gene family and of the V-J recombination. Eur J Immunol. 1992 Jun;22(6):1627–1634. doi: 10.1002/eji.1830220642. [DOI] [PubMed] [Google Scholar]
  18. Rada C., Gupta S. K., Gherardi E., Milstein C. Mutation and selection during the secondary response to 2-phenyloxazolone. Proc Natl Acad Sci U S A. 1991 Jul 1;88(13):5508–5512. doi: 10.1073/pnas.88.13.5508. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Reynaud C. A., Mackay C. R., Müller R. G., Weill J. C. Somatic generation of diversity in a mammalian primary lymphoid organ: the sheep ileal Peyer's patches. Cell. 1991 Mar 8;64(5):995–1005. doi: 10.1016/0092-8674(91)90323-q. [DOI] [PubMed] [Google Scholar]
  20. Rogerson B., Hackett J., Jr, Peters A., Haasch D., Storb U. Mutation pattern of immunoglobulin transgenes is compatible with a model of somatic hypermutation in which targeting of the mutator is linked to the direction of DNA replication. EMBO J. 1991 Dec;10(13):4331–4341. doi: 10.1002/j.1460-2075.1991.tb05011.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Rose M. L., Birbeck M. S., Wallis V. J., Forrester J. A., Davies A. J. Peanut lectin binding properties of germinal centres of mouse lymphoid tissue. Nature. 1980 Mar 27;284(5754):364–366. doi: 10.1038/284364a0. [DOI] [PubMed] [Google Scholar]
  22. 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]
  23. Sharpe M. J., Milstein C., Jarvis J. M., Neuberger M. S. Somatic hypermutation of immunoglobulin kappa may depend on sequences 3' of C kappa and occurs on passenger transgenes. EMBO J. 1991 Aug;10(8):2139–2145. doi: 10.1002/j.1460-2075.1991.tb07748.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Sharpe M. J., Neuberger M., Pannell R., Surani M. A., Milstein C. Lack of somatic mutation in a kappa light chain transgene. Eur J Immunol. 1990 Jun;20(6):1379–1385. doi: 10.1002/eji.1830200625. [DOI] [PubMed] [Google Scholar]
  25. Sohn J., Gerstein R. M., Hsieh C. L., Lemer M., Selsing E. Somatic hypermutation of an immunoglobulin mu heavy chain transgene. J Exp Med. 1993 Feb 1;177(2):493–504. doi: 10.1084/jem.177.2.493. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Weinstein P. D., Schweitzer P. A., Cebra-Thomas J. A., Cebra J. J. Molecular genetic features reflecting the preference for isotype switching to IgA expression by Peyer's patch germinal center B cells. Int Immunol. 1991 Dec;3(12):1253–1263. doi: 10.1093/intimm/3.12.1253. [DOI] [PubMed] [Google Scholar]