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. 1989 Apr;86(7):2346–2350. doi: 10.1073/pnas.86.7.2346

Isotype switching by a microinjected mu immunoglobulin heavy chain gene in transgenic mice.

J Durdik 1, R M Gerstein 1, S Rath 1, P F Robbins 1, A Nisonoff 1, E Selsing 1
PMCID: PMC286909  PMID: 2494666

Abstract

Immunization of transgenic mice carrying an immunoglobulin mu heavy chain resulted in a response dominated by expression of the transgene variable region. Unexpectedly, in a large proportion of the antibody produced by immunized mice, the transgene variable region was associated with IgG rather than IgM. This demonstrates that the transgene can undergo an isotype switch. Four transgenic founder lines all exhibited transgene isotype switching despite the likelihood of random chromosomal integration of the transgene. In addition one of the lines was analyzed by breeding studies and the transgene was found to be genetically unlinked to the immunoglobulin heavy chain (Igh) locus. These results indicate that a precise chromosomal location is not required for isotype switching and suggest the possibility that the isotype switching process can occur interchromosomally.

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Selected References

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

  1. Alt F. W., Blackwell T. K., Yancopoulos G. D. Development of the primary antibody repertoire. Science. 1987 Nov 20;238(4830):1079–1087. doi: 10.1126/science.3317825. [DOI] [PubMed] [Google Scholar]
  2. 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]
  3. Brinster R. L., Ritchie K. A., Hammer R. E., O'Brien R. L., Arp B., Storb U. Expression of a microinjected immunoglobulin gene in the spleen of transgenic mice. Nature. 1983 Nov 24;306(5941):332–336. doi: 10.1038/306332a0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Busto P., Gerstein R., Dupre L., Giorgetti C. A., Selsing E., Press J. L. Molecular analysis of heavy and light chains used by primary and secondary anti-(T,G)-A--L antibodies produced by normal and xid mice. J Immunol. 1987 Jul 15;139(2):608–618. [PubMed] [Google Scholar]
  5. Chen Y. W., Word C., Dev V., Uhr J. W., Vitetta E. S., Tucker P. W. Double isotype production by a neoplastic B cell line. II. Allelically excluded production of mu and gamma 1 heavy chains without CH gene rearrangement. J Exp Med. 1986 Aug 1;164(2):562–579. doi: 10.1084/jem.164.2.562. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cory S., Adams J. M. Deletions are associated with somatic rearrangement of immunoglobulin heavy chain genes. Cell. 1980 Jan;19(1):37–51. doi: 10.1016/0092-8674(80)90386-4. [DOI] [PubMed] [Google Scholar]
  7. Costantini F., Lacy E. Introduction of a rabbit beta-globin gene into the mouse germ line. Nature. 1981 Nov 5;294(5836):92–94. doi: 10.1038/294092a0. [DOI] [PubMed] [Google Scholar]
  8. 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]
  9. Davis M. M., Kim S. K., Hood L. E. DNA sequences mediating class switching in alpha-immunoglobulins. Science. 1980 Sep 19;209(4463):1360–1365. doi: 10.1126/science.6774415. [DOI] [PubMed] [Google Scholar]
  10. Dohi Y., Nisonoff A. Suppression of idiotype and generation of suppressor T cells with idiotype-conjugated thymocytes. J Exp Med. 1979 Oct 1;150(4):909–918. doi: 10.1084/jem.150.4.909. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Grosschedl R., Weaver D., Baltimore D., Costantini F. Introduction of a mu immunoglobulin gene into the mouse germ line: specific expression in lymphoid cells and synthesis of functional antibody. Cell. 1984 Oct;38(3):647–658. doi: 10.1016/0092-8674(84)90259-9. [DOI] [PubMed] [Google Scholar]
  12. Hamlyn P. H., Browniee G. G., Cheng C. C., Gait M. J., Milstein C. Complete sequence of constant and 3' noncoding regions of an immunoglobulin mRNA using the dideoxynucleotide method of RNA sequencing. Cell. 1978 Nov;15(3):1067–1075. doi: 10.1016/0092-8674(78)90290-8. [DOI] [PubMed] [Google Scholar]
  13. Hornbeck P. V., Lewis G. K. Idiotype connectance in the immune system. I. Expression of a cross-reactive idiotype on induced anti-p-azophenylarsonate antibodies and on endogenous antibodies not specific for arsonate. J Exp Med. 1983 Apr 1;157(4):1116–1136. doi: 10.1084/jem.157.4.1116. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Jäck H. M., McDowell M., Steinberg C. M., Wabl M. Looping out and deletion mechanism for the immunoglobulin heavy-chain class switch. Proc Natl Acad Sci U S A. 1988 Mar;85(5):1581–1585. doi: 10.1073/pnas.85.5.1581. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kataoka T., Kawakami T., Takahashi N., Honjo T. Rearrangement of immunoglobulin gamma 1-chain gene and mechanism for heavy-chain class switch. Proc Natl Acad Sci U S A. 1980 Feb;77(2):919–923. doi: 10.1073/pnas.77.2.919. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Kipps T. J., Herzenberg L. A. Homologous chromosome recombination generating immunoglobulin allotype and isotype switch variants. EMBO J. 1986 Feb;5(2):263–268. doi: 10.1002/j.1460-2075.1986.tb04208.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Knight K. L., Malek T. R., Hanly W. C. Recombinant rabbit secretory immunoglobulin molecules: alpha chains with maternal (paternal) variable-region allotypes and paternal (maternal) constant-region allotypes. Proc Natl Acad Sci U S A. 1974 Apr;71(4):1169–1173. doi: 10.1073/pnas.71.4.1169. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Kuettner M. G., Wang A. L., Nisonoff A. Quantitative investigations of idiotypic antibodies. VI. Idiotypic specificity as a potential genetic marker for the variable regions of mouse immunoglobulin polypeptide chains. J Exp Med. 1972 Mar 1;135(3):579–595. doi: 10.1084/jem.135.3.579. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Lacy E., Roberts S., Evans E. P., Burtenshaw M. D., Costantini F. D. A foreign beta-globin gene in transgenic mice: integration at abnormal chromosomal positions and expression in inappropriate tissues. Cell. 1983 Sep;34(2):343–358. doi: 10.1016/0092-8674(83)90369-0. [DOI] [PubMed] [Google Scholar]
  20. Lamoyi E., Estess P., Capra J. D., Nisonoff A. Heterogeneity of an intrastrain cross-reactive idiotype associated with anti-p-azophenylarsonate antibodies of A/J mice. J Immunol. 1980 Jun;124(6):2834–2840. [PubMed] [Google Scholar]
  21. Laskin J. A., Gray A., Nisonoff A., Klinman N. R., Gottlieb P. D. Segregation at a locus determining an immunoglobulin genetic marker for the light chain variable region affects inheritance of expression of an idiotype. Proc Natl Acad Sci U S A. 1977 Oct;74(10):4600–4604. doi: 10.1073/pnas.74.10.4600. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Maki R., Traunecker A., Sakano H., Roeder W., Tonegawa S. Exon shuffling generates an immunoglobulin heavy chain gene. Proc Natl Acad Sci U S A. 1980 Apr;77(4):2138–2142. doi: 10.1073/pnas.77.4.2138. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Marcu K. B., Banerji J., Penncavage N. A., Lang R., Arnheim N. 5' flanking region of immunoglobulin heavy chain constant region genes displays length heterogeneity in germlines of inbred mouse strains. Cell. 1980 Nov;22(1 Pt 1):187–196. doi: 10.1016/0092-8674(80)90167-1. [DOI] [PubMed] [Google Scholar]
  24. O'Brien R. L., Brinster R. L., Storb U. Somatic hypermutation of an immunoglobulin transgene in kappa transgenic mice. 1987 Mar 26-Apr 1Nature. 326(6111):405–409. doi: 10.1038/326405a0. [DOI] [PubMed] [Google Scholar]
  25. Obata M., Kataoka T., Nakai S., Yamagishi H., Takahashi N., Yamawaki-Kataoka Y., Nikaido T., Shimizu A., Honjo T. Structure of a rearranged gamma 1 chain gene and its implication to immunoglobulin class-switch mechanism. Proc Natl Acad Sci U S A. 1981 Apr;78(4):2437–2441. doi: 10.1073/pnas.78.4.2437. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Perlmutter A. P., Gilbert W. Antibodies of the secondary response can be expressed without switch recombination in normal mouse B cells. Proc Natl Acad Sci U S A. 1984 Nov;81(22):7189–7193. doi: 10.1073/pnas.81.22.7189. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Rabbitts T. H., Forster A., Dunnick W., Bentley D. L. The role of gene deletion in the immunoglobulin heavy chain switch. Nature. 1980 Jan 24;283(5745):351–356. doi: 10.1038/283351a0. [DOI] [PubMed] [Google Scholar]
  28. Radbruch A., Burger C., Klein S., Müller W. Control of immunoglobulin class switch recombination. Immunol Rev. 1986 Feb;89:69–83. doi: 10.1111/j.1600-065x.1986.tb01473.x. [DOI] [PubMed] [Google Scholar]
  29. Robbins P. F., Rosen E. M., Haba S., Nisonoff A. Relationship of VH and VL genes encoding three idiotypic families of anti-p-azobenzenearsonate antibodies. Proc Natl Acad Sci U S A. 1986 Feb;83(4):1050–1054. doi: 10.1073/pnas.83.4.1050. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Siekevitz M., Gefter M. L., Brodeur P., Riblet R., Marshak-Rothstein A. The genetic basis of antibody production: the dominant anti-arsonate idiotype response of the strain A mouse. Eur J Immunol. 1982 Dec;12(12):1023–1032. doi: 10.1002/eji.1830121208. [DOI] [PubMed] [Google Scholar]
  31. 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]
  32. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  33. Storb U., Pinkert C., Arp B., Engler P., Gollahon K., Manz J., Brady W., Brinster R. L. Transgenic mice with mu and kappa genes encoding antiphosphorylcholine antibodies. J Exp Med. 1986 Aug 1;164(2):627–641. doi: 10.1084/jem.164.2.627. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Tilley S. A., Birshtein B. K. Unequal sister chromatid exchange. A mechanism affecting Ig gene arrangement and expression. J Exp Med. 1985 Aug 1;162(2):675–694. doi: 10.1084/jem.162.2.675. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Wabl M., Meyer J., Beck-Engeser G., Tenkhoff M., Burrows P. D. Critical test of a sister chromatid exchange model for the immunoglobulin heavy-chain class switch. Nature. 1985 Feb 21;313(6004):687–689. doi: 10.1038/313687a0. [DOI] [PubMed] [Google Scholar]
  36. Weaver D., Reis M. H., Albanese C., Costantini F., Baltimore D., Imanishi-Kari T. Altered repertoire of endogenous immunoglobulin gene expression in transgenic mice containing a rearranged mu heavy chain gene. Cell. 1986 Apr 25;45(2):247–259. doi: 10.1016/0092-8674(86)90389-2. [DOI] [PubMed] [Google Scholar]
  37. Wysocki L. J., Gridley T., Huang S., Grandea A. G., 3rd, Gefter M. L. Single germline VH and V kappa genes encode predominating antibody variable regions elicited in strain A mice by immunization with p-azophenylarsonate. J Exp Med. 1987 Jul 1;166(1):1–11. doi: 10.1084/jem.166.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Yaoita Y., Kumagai Y., Okumura K., Honjo T. Expression of lymphocyte surface IgE does not require switch recombination. Nature. 1982 Jun 24;297(5868):697–699. doi: 10.1038/297697a0. [DOI] [PubMed] [Google Scholar]

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