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. 1996 Nov;89(3):356–362. doi: 10.1046/j.1365-2567.1996.d01-749.x

Lipopolysaccharide prevents apoptosis and induces responsiveness to antigen receptor cross-linking in immature B cells.

R J Wechsler-Reya 1, J G Monroe 1
PMCID: PMC1456554  PMID: 8958047

Abstract

Unlike mature B cells, immature B cells are not activated in response to antigen receptor cross-linking. To examine the mechanisms underlying this unresponsiveness, we have studied the effects of reagents that have been shown to alter the responses of immature B cells to antigen receptor stimulation. Bacterial lipopolysaccharide (LPS) is a polyclonal B-cell activator, and has been shown to interfere with B-cell tolerance induction in vivo and in vitro. Here we show that LPS can also overcome the unresponsiveness of immature B cells to stimulation with anti-receptor (anti-mu) antibodies. LPS synergizes with anti-mu to induce a proliferative response that exceeds the response of immature B cells to LPS alone. Moreover, pretreatment of immature cells with LPS allows them to proliferate in response to subsequent stimulation with anti-mu antibodies. This induction of responsiveness to anti-mu requires exposure to LPS for at least 8 hr. Although the mechanisms of induction are not fully understood, one component of the LPS effect appears to involve enhancement of immature B-cell survival in culture. Neonatal splenic B cells undergo spontaneous apoptosis at a much higher rate than mature B cells, but we have found that LPS causes a dramatic inhibition of apoptosis, even when it is present for only the first 8 hr of culture. The ability of LPS to promote survival of immature B cells and allow them to proliferate in response to antigen receptor stimulation provides a system for investigation of the biochemical mechanisms of unresponsiveness and tolerance susceptibility.

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Selected References

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

  1. Adams E., Basten A., Goodnow C. C. Intrinsic B-cell hyporesponsiveness accounts for self-tolerance in lysozyme/anti-lysozyme double-transgenic mice. Proc Natl Acad Sci U S A. 1990 Aug;87(15):5687–5691. doi: 10.1073/pnas.87.15.5687. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Austyn J. M., Gordon S. F4/80, a monoclonal antibody directed specifically against the mouse macrophage. Eur J Immunol. 1981 Oct;11(10):805–815. doi: 10.1002/eji.1830111013. [DOI] [PubMed] [Google Scholar]
  3. Benhamou L. E., Cazenave P. A., Sarthou P. Anti-immunoglobulins induce death by apoptosis in WEHI-231 B lymphoma cells. Eur J Immunol. 1990 Jun;20(6):1405–1407. doi: 10.1002/eji.1830200630. [DOI] [PubMed] [Google Scholar]
  4. Boscá L., Márquez C., Martínez C. B cell triggering by bacterial lipopeptide involves both translocation and activation of the membrane-bound form of protein kinase C. J Immunol. 1991 Sep 1;147(5):1463–1469. [PubMed] [Google Scholar]
  5. Brines R. D., Klaus G. G. Inhibition of lipopolysaccharide-induced activation of immature B cells by anti-mu and anti-delta antibodies and its modulation by interleukin-4. Int Immunol. 1992 Jul;4(7):765–771. doi: 10.1093/intimm/4.7.765. [DOI] [PubMed] [Google Scholar]
  6. Carman J. A., Wechsler-Reya R. J., Monroe J. G. Immature stage B cells enter but do not progress beyond the early G1 phase of the cell cycle in response to antigen receptor signaling. J Immunol. 1996 Jun 15;156(12):4562–4569. [PubMed] [Google Scholar]
  7. Chang T. L., Capraro G., Kleinman R. E., Abbas A. K. Anergy in immature B lymphocytes. Differential responses to receptor-mediated stimulation and T helper cells. J Immunol. 1991 Aug 1;147(3):750–756. [PubMed] [Google Scholar]
  8. Gupta S., Gollapudi S., Vayuvegula B. Lipopolysaccharide-induced murine B cell proliferation: a role of protein kinase C. Cell Immunol. 1988 Dec;117(2):425–429. doi: 10.1016/0008-8749(88)90131-1. [DOI] [PubMed] [Google Scholar]
  9. Hardy R. R., Carmack C. E., Shinton S. A., Kemp J. D., Hayakawa K. Resolution and characterization of pro-B and pre-pro-B cell stages in normal mouse bone marrow. J Exp Med. 1991 May 1;173(5):1213–1225. doi: 10.1084/jem.173.5.1213. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hardy R. R. Variable gene usage, physiology and development of Ly-1+ (CD5+) B cells. Curr Opin Immunol. 1992 Apr;4(2):181–185. doi: 10.1016/0952-7915(92)90010-c. [DOI] [PubMed] [Google Scholar]
  11. Hasbold J., Klaus G. G. Anti-immunoglobulin antibodies induce apoptosis in immature B cell lymphomas. Eur J Immunol. 1990 Aug;20(8):1685–1690. doi: 10.1002/eji.1830200810. [DOI] [PubMed] [Google Scholar]
  12. Holmes K. L., Langdon W. Y., Fredrickson T. N., Coffman R. L., Hoffman P. M., Hartley J. W., Morse H. C., 3rd Analysis of neoplasms induced by Cas-Br-M MuLV tumor extracts. J Immunol. 1986 Jul 15;137(2):679–688. [PubMed] [Google Scholar]
  13. Illera V. A., Perandones C. E., Stunz L. L., Mower D. A., Jr, Ashman R. F. Apoptosis in splenic B lymphocytes. Regulation by protein kinase C and IL-4. J Immunol. 1993 Sep 15;151(6):2965–2973. [PubMed] [Google Scholar]
  14. Jakway J. P., Usinger W. R., Gold M. R., Mishell R. I., DeFranco A. L. Growth regulation of the B lymphoma cell line WEHI-231 by anti-immunoglobulin, lipopolysaccharide, and other bacterial products. J Immunol. 1986 Oct 1;137(7):2225–2231. [PubMed] [Google Scholar]
  15. Kamesaki H., Zwiebel J. A., Reed J. C., Cossman J. Role of bcl-2 and IL-5 in the regulation of anti-IgM-induced growth arrest and apoptosis in immature B cell lines. A cooperative regulation model for B cell clonal deletion. J Immunol. 1994 Apr 1;152(7):3294–3305. [PubMed] [Google Scholar]
  16. Kearney J. F., Cooper M. D., Lawton A. R. B lymphocyte differentiation induced by lipopolysaccharide. III. Suppression of B cell maturation by anti-mouse immunoglobulin antibodies. J Immunol. 1976 Jun;116(6):1664–1668. [PubMed] [Google Scholar]
  17. Kearney J. F., Lawton A. R. B lymphocyte differentiation induced by lipopolysaccharide. I. Generation of cells synthesizing four major immunoglobulin classes. J Immunol. 1975 Sep;115(3):671–676. [PubMed] [Google Scholar]
  18. Kruger M. G., Riley R. L., Riley E. A., Elia J. M. Bone marrow stromal cells modulate both kappa light chain and Ly1 antigen expression on Ly1+ pre-B cell lines in vitro. Blood. 1990 Jul 15;76(2):383–392. [PubMed] [Google Scholar]
  19. Lanier L. L., Gutman G. A., Lewis D. E., Griswold S. T., Warner N. L. Monoclonal antibodies against rat immunoglobulin kappa chains. Hybridoma. 1982;1(2):125–131. doi: 10.1089/hyb.1.1982.1.125. [DOI] [PubMed] [Google Scholar]
  20. Louis J. A., Chiller J. M., Weigle W. O. The ability of bacterial lipopolysaccharide to modulate the induction of unresponsiveness to a state of immunity. Cellular parameters. J Exp Med. 1973 Dec 1;138(6):1481–1495. doi: 10.1084/jem.138.6.1481. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Marshak-Rothstein A., Fink P., Gridley T., Raulet D. H., Bevan M. J., Gefter M. L. Properties and applications of monoclonal antibodies directed against determinants of the Thy-1 locus. J Immunol. 1979 Jun;122(6):2491–2497. [PubMed] [Google Scholar]
  22. Metcalf E. S., Klinman N. R. In vitro tolerance induction of neonatal murine B cells. J Exp Med. 1976 Jun 1;143(6):1327–1340. doi: 10.1084/jem.143.6.1327. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Norvell A., Mandik L., Monroe J. G. Engagement of the antigen-receptor on immature murine B lymphocytes results in death by apoptosis. J Immunol. 1995 May 1;154(9):4404–4413. [PubMed] [Google Scholar]
  24. Nossal G. J. Negative selection of lymphocytes. Cell. 1994 Jan 28;76(2):229–239. doi: 10.1016/0092-8674(94)90331-x. [DOI] [PubMed] [Google Scholar]
  25. Nossal G. J., Pike B. L. Mechanisms of clonal abortion tolerogenesis. I. Response of immature hapten-specific B lymphocytes. J Exp Med. 1978 Nov 1;148(5):1161–1170. doi: 10.1084/jem.148.5.1161. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Ornellas E. P., Sanfilippo F., Scott D. W. Cellular events in tolerance. IV. The effect of a graft-versus-host reaction and endotoxin on hapten- and carrier-specific tolerance. Eur J Immunol. 1974 Sep;4(9):587–591. doi: 10.1002/eji.1830040902. [DOI] [PubMed] [Google Scholar]
  27. Paige C. J., Kincade P. W., Ralph P. Murine B cell leukemia line with inducible surface immunoglobulin expression. J Immunol. 1978 Aug;121(2):641–647. [PubMed] [Google Scholar]
  28. Pike B. L., Boyd A. W., Nossal G. J. Clonal anergy: the universally anergic B lymphocyte. Proc Natl Acad Sci U S A. 1982 Mar;79(6):2013–2017. doi: 10.1073/pnas.79.6.2013. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Raetz C. R. Biochemistry of endotoxins. Annu Rev Biochem. 1990;59:129–170. doi: 10.1146/annurev.bi.59.070190.001021. [DOI] [PubMed] [Google Scholar]
  30. Raff M. C., Owen J. J., Cooper M. D., Lawton A. R., 3rd, Megson M., Gathings W. E. Differences in susceptibility of mature and immature mouse B lymphocytes to anti-immunoglobulin-induced immunoglobulin suppression in vitro. Possible implications for B-cell tolerance to self. J Exp Med. 1975 Nov 1;142(5):1052–1064. doi: 10.1084/jem.142.5.1052. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Sieckmann D. G., Scher I., Asofsky R., Mosier D. E., Paul W. E. Activation of mouse lymphocytes by anti-immunoglobulin. II. A thymus-independent response by a mature subset of B lymphocytes. J Exp Med. 1978 Dec 1;148(6):1628–1643. doi: 10.1084/jem.148.6.1628. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Sitia R., Abbott J., Hämmerling U. The ontogeny of B lymphocytes. V. Lipopolysaccharide-induced changes of IgD expression on murine B lymphocytes. Eur J Immunol. 1979 Nov;9(11):859–864. doi: 10.1002/eji.1830091106. [DOI] [PubMed] [Google Scholar]
  33. Skidmore B. J., Chiller J. M., Morrison D. C., Weigle W. O. Immunologic properties of bacterial lipopolysaccharide (LPS): correlation between the mitogenic, adjuvant, and immunogenic activities. J Immunol. 1975 Feb;114(2 Pt 2):770–775. [PubMed] [Google Scholar]
  34. Stewart B. W. Mechanisms of apoptosis: integration of genetic, biochemical, and cellular indicators. J Natl Cancer Inst. 1994 Sep 7;86(17):1286–1296. doi: 10.1093/jnci/86.17.1286. [DOI] [PubMed] [Google Scholar]
  35. Su S. D., Ward M. M., Apicella M. A., Ward R. E. The primary B cell response to the O/core region of bacterial lipopolysaccharide is restricted to the Ly-1 lineage. J Immunol. 1991 Jan 1;146(1):327–331. [PubMed] [Google Scholar]
  36. Wechsler R. J., Monroe J. G. Immature B lymphocytes are deficient in expression of the src-family kinases p59fyn and p55fgr1. J Immunol. 1995 Feb 15;154(4):1919–1929. [PubMed] [Google Scholar]
  37. Yellen-Shaw A., Monroe J. G. Differential responsiveness of immature- and mature-stage murine B cells to anti-IgM reflects both FcR-dependent and -independent mechanisms. Cell Immunol. 1992 Dec;145(2):339–350. doi: 10.1016/0008-8749(92)90336-n. [DOI] [PubMed] [Google Scholar]
  38. Yellen A. J., Glenn W., Sukhatme V. P., Cao X. M., Monroe J. G. Signaling through surface IgM in tolerance-susceptible immature murine B lymphocytes. Developmentally regulated differences in transmembrane signaling in splenic B cells from adult and neonatal mice. J Immunol. 1991 Mar 1;146(5):1446–1454. [PubMed] [Google Scholar]
  39. Zanetti M. Ontogeny of the immune system and the invisible frontier to immune regulation. Int Rev Immunol. 1992;8(2-3):209–218. doi: 10.3109/08830189209055574. [DOI] [PubMed] [Google Scholar]

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