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. 1979 Jul;76(7):3469–3473. doi: 10.1073/pnas.76.7.3469

Assembly and secretion of pentameric IgM in a fusion between a nonsecreting B cell lymphoma and an IgG-secreting plasmacytoma.

W C Raschke, E L Mather, M E Koshland
PMCID: PMC383847  PMID: 115002

Abstract

A new immunoglobulin product has been obtained by hybridization of mouse cell lines arrested at different stages in B lymphocyte development. One line was shown to have the characteristics of an undifferentiated B cell that synthesizes monomeric IgM as a membrane receptor but does not express J chain. The second line was represent of a fully differentiated plasma cell synthesizing large amounts of IgG and J chain, but no IgM. Fusion of the two cell types yielded independent hybrid clones that secreted pentameric IgM, normally the first product of antigen-driven B cell differentiation. Analyses of the hybrid cells indicated that the IgM was expressed as a result of complementation between the synthetic capacities of the parental lines. The hybrid cells synthesized both monomeric IgM and J chain and assembled these components into a pentameric molecule with the expected stoichiometry of one J chain per five monomeric units. These findings provided further evidence that the induction of B cell differentiation includes a signal for de novo synthesis of the J chain. Moreover, the complementation achieved by this hybridization provides a system for identifying other intracellular events in B cell differentiation to IgM secretion.

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Selected References

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  1. Andersson J., Buxbaum J., Citronbaum R., Douglas S., Forni L., Melchers F., Pernis B., Stott D. IgM-producing tumors in the BALB-c mouse: a model for B-cell maturation. J Exp Med. 1974 Sep 1;140(3):742–763. doi: 10.1084/jem.140.3.742. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Andersson J., Sjöberg O., Möller G. Mitogens as probes for immunocyte activation and cellular cooperation. Transplant Rev. 1972;11:131–177. doi: 10.1111/j.1600-065x.1972.tb00048.x. [DOI] [PubMed] [Google Scholar]
  3. Bergman Y., Haimovich J., Melchers F. An IgM-producing tumor with biochemical characteristics of a small B lymphocyte. Eur J Immunol. 1977 Aug;7(8):574–579. doi: 10.1002/eji.1830070815. [DOI] [PubMed] [Google Scholar]
  4. Bonner W. M., Stedman J. D. Efficient fluorography of 3H and 14C on thin layers. Anal Biochem. 1978 Aug 15;89(1):247–256. doi: 10.1016/0003-2697(78)90747-9. [DOI] [PubMed] [Google Scholar]
  5. Brown J. C., Koshland M. E. Evidence for a long-range conformational change induced by antigen binding to IgM antibody. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5682–5686. doi: 10.1073/pnas.74.12.5682. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Crissman H. A., Tobey R. A. Cell-cycle analysis in 20 minutes. Science. 1974 Jun 21;184(4143):1297–1298. doi: 10.1126/science.184.4143.1297. [DOI] [PubMed] [Google Scholar]
  7. Davidson R. L. Gene expression in somatic cell hybrids. Annu Rev Genet. 1974;8:195–218. doi: 10.1146/annurev.ge.08.120174.001211. [DOI] [PubMed] [Google Scholar]
  8. Dennert G., Raschke W. Continuously proliferating allospecific T cells, lifespan and antigen receptors. Eur J Immunol. 1977 Jun;7(6):352–359. doi: 10.1002/eji.1830070606. [DOI] [PubMed] [Google Scholar]
  9. Dingman C. W., Peacock A. C. Analytical studies on nuclear ribonucleic acid using polyacrylamide gel electrophoresis. Biochemistry. 1968 Feb;7(2):659–668. doi: 10.1021/bi00842a022. [DOI] [PubMed] [Google Scholar]
  10. Greaves M., Janossy G. Elicitation of selective T and B lymphocyte responses by cell surface binding ligands. Transplant Rev. 1972;11:87–130. doi: 10.1111/j.1600-065x.1972.tb00047.x. [DOI] [PubMed] [Google Scholar]
  11. Grey H. M., Colón S., Campbell P., Rabellino E. Immunoglobulins on the surface of lymphocytes. V. Quantitative studies on the question of whether immunoglobulins are associated with T cells in the mouse. J Immunol. 1972 Oct;109(4):776–783. [PubMed] [Google Scholar]
  12. Hutchinson M. A., Hunter T., Eckhart W. Characterization of T antigens in polyoma-infected and transformed cells. Cell. 1978 Sep;15(1):65–77. doi: 10.1016/0092-8674(78)90083-1. [DOI] [PubMed] [Google Scholar]
  13. Hyman R., Stallings V. Complementation patterns of Thy-1 variants and evidence that antigen loss variants "pre-exist" in the parental population. J Natl Cancer Inst. 1974 Feb;52(2):429–436. doi: 10.1093/jnci/52.2.429. [DOI] [PubMed] [Google Scholar]
  14. Kaji H., Parkhouse R. M. Intracellular J chain in mouse plasmacytomas secreting IgA, IgM and IgG. Nature. 1974 May 3;249(452):45–47. doi: 10.1038/249045a0. [DOI] [PubMed] [Google Scholar]
  15. Kehry M., Sibley C., Fuhrman J., Schilling J., Hood L. E. Amino acid sequence of a mouse immunoglobulin mu chain. Proc Natl Acad Sci U S A. 1979 Jun;76(6):2932–2936. doi: 10.1073/pnas.76.6.2932. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Kessler S. W. Rapid isolation of antigens from cells with a staphylococcal protein A-antibody adsorbent: parameters of the interaction of antibody-antigen complexes with protein A. J Immunol. 1975 Dec;115(6):1617–1624. [PubMed] [Google Scholar]
  17. Koshland M. E. Structure and function of the J chain. Adv Immunol. 1975;20:41–69. doi: 10.1016/s0065-2776(08)60206-0. [DOI] [PubMed] [Google Scholar]
  18. Köhler G., Howe S. C., Milstein C. Fusion between immunoglobulin-secreting and nonsecreting myeloma cell lines. Eur J Immunol. 1976 Apr;6(4):292–295. doi: 10.1002/eji.1830060411. [DOI] [PubMed] [Google Scholar]
  19. Levy R., Dilley J. Rescue of immunoglobulin secretion from human neoplastic lymphoid cells by somatic cell hybridization. Proc Natl Acad Sci U S A. 1978 May;75(5):2411–2415. doi: 10.1073/pnas.75.5.2411. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Marchalonis J. J. An enzymic method for the trace iodination of immunoglobulins and other proteins. Biochem J. 1969 Jun;113(2):299–305. doi: 10.1042/bj1130299. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Margulies D. H., Kuehl W. M., Scharff M. D. Somatic cell hybridization of mouse myeloma cells. Cell. 1976 Jul;8(3):405–415. doi: 10.1016/0092-8674(76)90153-7. [DOI] [PubMed] [Google Scholar]
  22. Melchers F., Andersson J. IgM in bone marrow-derived lymphocytes. Changes in synthesis, turnover and secretion, and in numbers of molecules on the surface of B cells after mitogenic stimulation. Eur J Immunol. 1974 Mar;4(3):181–188. doi: 10.1002/eji.1830040306. [DOI] [PubMed] [Google Scholar]
  23. Melchers F. Difference in carbohydrate composition and a possible conformational difference between intracellular and extracellular immunoglobulin M. Biochemistry. 1972 May 23;11(11):2204–2208. doi: 10.1021/bi00761a031. [DOI] [PubMed] [Google Scholar]
  24. Mosier D. E. A requirement for two cell types for antibody formation in vitro. Science. 1967 Dec 22;158(3808):1573–1575. doi: 10.1126/science.158.3808.1573. [DOI] [PubMed] [Google Scholar]
  25. Mosmann T., Baumal R. Synthesis but not secretion of J chain by variant mouse myeloma cells which lose alpha-chain-synthesizing ability. J Immunol. 1975 Oct;115(4):955–962. [PubMed] [Google Scholar]
  26. POTTER M., FAHEY J. L. Studies on eight transplantable plasma-cell neoplasms of mice. J Natl Cancer Inst. 1960 May;24:1153–1165. [PubMed] [Google Scholar]
  27. Raschke W. C. Expression of murine IgM, IgD and Ia molecules on hybrids of murine LPS blasts with a Syrian hamster B lymphoma. Curr Top Microbiol Immunol. 1978;81:70–76. doi: 10.1007/978-3-642-67448-8_12. [DOI] [PubMed] [Google Scholar]
  28. Reisfeld R. A., Small P. A., Jr Electrophoretic heterogeneity of polypeptide chains of specific antibodies. Science. 1966 May 27;152(3726):1253–1255. doi: 10.1126/science.152.3726.1253. [DOI] [PubMed] [Google Scholar]
  29. Sekiguchi T., Sekiguchi F. Interallelic complementation in hybrid cells derived from Chinese hamster diploid clones deficient in hypoxanthine-guanine phosphoribosyl-transferase activity. Exp Cell Res. 1973 Mar 15;77(1):391–403. doi: 10.1016/0014-4827(73)90593-4. [DOI] [PubMed] [Google Scholar]
  30. Smith G. P. Sequence of the full-length immunoglobulin kappa-chain of mouse myeloma MPC 11. Biochem J. 1978 May 1;171(2):337–347. doi: 10.1042/bj1710337. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Wilde C. E., 3rd, Koshland M. E. Molecular size and shape of the J chain from polymeric immunoglobulins. Biochemistry. 1973 Aug 14;12(17):3218–3224. doi: 10.1021/bi00741a012. [DOI] [PubMed] [Google Scholar]

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