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CD45 links the B cell receptor with cell survival and is required for the persistence of germinal centers

Nature Immunology volume 7, pages 190–198 (2006)Cite this article

Abstract

To segregate the many contributions that B cell receptor (BCR)–mediated signals make to immune responses, we have analyzed here B cells deficient in the 'pan-leukocyte' marker CD45. BCR ligation of Cd45−/− B cells failed to activate phosphatidylinositol-3-OH kinase, NF-κB, Erk1 or Erk2 kinases or to upregulate cell survival proteins and instead induced apoptosis. Immunization of Cd45−/− B cell chimeras induced germinal centers and antigen-specific immunoglobulin G1 antibody-forming cells early, but both cellular compartments decreased by day 14. Proliferation of Cd45−/− B cells induced by CD40 ligand in vitro was impaired as a result of abrogation by BCR ligation of the upregulation of prosurvival proteins. In contrast, enforced expression of the antiapoptotic factor Bcl-xL prevented the collapse of Cd45−/− B cell germinal centers. These results show mechanistic differences in B cell survival during germinal center initiation and propagation; CD40 signaling is sufficient for the former, whereas the latter requires signaling from the BCR.

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Figure 1: Cellular responses after BCR ligation of Cd45−/− B cells.
Figure 2: CD45-null B cells respond normally to growth factors, have normal frequency of NP-reactive BCRs and efficiently present antigen.
Figure 3: B cell chimeras.
Figure 4: Reduced NP-IgG1+ AFC generation in the absence of CD45.
Figure 5: Premature dissolution of germinal centers and NP+IgG1+ cells in Cd45−/− B cell chimeras.
Figure 6: BCR ligation of Cd45−/− B cells inhibits CD40L-mediated proliferation and results in enhanced apoptosis in vitro.
Figure 7: Enforced expression of Bcl-xL permits Cd45−/− B cells to maintain T cell–dependent responses.

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References

  1. Cyster, J.G. Chemokines, sphingosine-1-phosphate, and cell migration in secondary lymphoid organs. Annu. Rev. Immunol. 23, 127–159 (2005).

    Article  CAS  Google Scholar 

  2. van Kooten, C. & Banchereau, J. CD40–CD40 ligand. J. Leukoc. Biol. 67, 2–17 (2000).

    Article  CAS  Google Scholar 

  3. Tarlinton, D. Germinal centers: form and function. Curr. Opin. Immunol. 10, 245–251 (1998).

    Article  CAS  Google Scholar 

  4. Wolniak, K.L., Shinall, S.M. & Waldschmidt, T.J. The germinal center response. Crit. Rev. Immunol. 24, 39–65 (2004).

    Article  CAS  Google Scholar 

  5. Liu, Y.J. et al. Mechanism of antigen-driven selection in germinal centres. Nature 342, 929–931 (1989).

    Article  CAS  Google Scholar 

  6. Lane, P. et al. Soluble CD40 ligand can replace the normal T cell–derived CD40 ligand signal to B cells in T cell–dependent activation. J. Exp. Med. 177, 1209–1213 (1993).

    Article  CAS  Google Scholar 

  7. Grillot, D.A. et al. Bcl-x exhibits regulated expression during B cell development and activation and modulates lymphocyte survival in transgenic mice. J. Exp. Med. 183, 381–391 (1996).

    Article  CAS  Google Scholar 

  8. Lanzavecchia, A. & Sallusto, F. Progressive differentiation and selection of the fittest in the immune response. Nat. Rev. Immunol. 2, 982–987 (2002).

    Article  CAS  Google Scholar 

  9. Thomas, M.L. The regulation of B- and T-lymphocyte activation by the transmembrane protein tyrosine phosphatase CD45. Curr. Opin. Cell Biol. 6, 247–252 (1994).

    Article  CAS  Google Scholar 

  10. Huntington, N.D. & Tarlinton, D.M. CD45: direct and indirect government of immune regulation. Immunol. Lett. 94, 167–174 (2004).

    Article  CAS  Google Scholar 

  11. Pani, G., Siminovitch, K.A. & Paige, C.J. The motheaten mutation rescues B cell signaling and development in CD45-deficient mice. J. Exp. Med. 186, 581–588 (1997).

    Article  CAS  Google Scholar 

  12. Kishihara, K. et al. Normal B lymphocyte development but impaired T cell maturation in CD45-exon6 protein tyrosine phosphatase-deficient mice. Cell 74, 143–156 (1993).

    Article  CAS  Google Scholar 

  13. Benatar, T. et al. Immunoglobulin-mediated signal transduction in B cells from CD45-deficient mice. J. Exp. Med. 183, 329–334 (1996).

    Article  CAS  Google Scholar 

  14. Cyster, J.G. et al. Regulation of B-lymphocyte negative and positive selection by tyrosine phosphatase CD45. Nature 381, 325–328 (1996).

    Article  CAS  Google Scholar 

  15. Byth, K.F. et al. CD45-null transgenic mice reveal a positive regulatory role for CD45 in early thymocyte development, in the selection of CD4+CD8+ thymocytes, and B cell maturation. J. Exp. Med. 183, 1707–1718 (1996).

    Article  CAS  Google Scholar 

  16. Kong, Y.Y. et al. Differential requirements of CD45 for lymphocyte development and function. Eur. J. Immunol. 25, 3431–3436 (1995).

    Article  CAS  Google Scholar 

  17. Mee, P.J. et al. Greatly reduced efficiency of both positive and negative selection of thymocytes in CD45 tyrosine phosphatase-deficient mice. Eur. J. Immunol. 29, 2923–2933 (1999).

    Article  CAS  Google Scholar 

  18. Niiro, H. & Clark, E.A. Regulation of B-cell fate by antigen-receptor signals. Nat. Rev. Immunol. 2, 945–956 (2002).

    Article  CAS  Google Scholar 

  19. Baeuerle, P.A. & Baltimore, D. NF-κB: ten years after. Cell 87, 13–20 (1996).

    Article  CAS  Google Scholar 

  20. Grumont, R.J., Rourke, I.J. & Gerondakis, S. Rel-dependent induction of A1 transcription is required to protect B cells from antigen receptor ligation-induced apoptosis. Genes Dev. 13, 400–411 (1999).

    Article  CAS  Google Scholar 

  21. Allen, D. et al. Timing, genetic requirements and functional consequences of somatic hypermutation during B-cell development. Immunol. Rev. 96, 5–22 (1987).

    Article  CAS  Google Scholar 

  22. Rahman, Z.S., Rao, S.P., Kalled, S.L. & Manser, T. Normal induction but attenuated progression of germinal center responses in BAFF and BAFF-R signaling-deficient mice. J. Exp. Med. 198, 1157–1169 (2003).

    Article  CAS  Google Scholar 

  23. Kitamura, D., Roes, J., Kuhn, R. & Rajewsky, K.A. B cell-deficient mouse by targeted disruption of the membrane exon of the immunoglobulin μ chain gene. Nature 350, 423–426 (1991).

    Article  CAS  Google Scholar 

  24. MacLennan, I.C. Germinal centers. Annu. Rev. Immunol. 12, 117–139 (1994).

    Article  CAS  Google Scholar 

  25. Van Parijs, L., Refaeli, Y., Abbas, A.K. & Baltimore, D. Autoimmunity as a consequence of retrovirus-mediated expression of C-FLIP in lymphocytes. Immunity 11, 763–770 (1999).

    Article  CAS  Google Scholar 

  26. Rickert, R.C., Rajewsky, K. & Roes, J. Impairment of T cell–dependent B cell responses and B-1 cell development in CD19-deficient mice. Nature 376, 352–355 (1995).

    Article  CAS  Google Scholar 

  27. Ridderstad, A. & Tarlinton, D.M. Kinetics of establishing the memory B cell population as revealed by CD38 expression. J. Immunol. 160, 4688–4695 (1998).

    CAS  PubMed  Google Scholar 

  28. Xu, Y., Beavitt, S.J., Harder, K.W., Hibbs, M.L. & Tarlinton, D.M. The activation and subsequent regulatory roles of Lyn and CD19 after B cell receptor ligation are independent. J. Immunol. 169, 6910–6918 (2002).

    Article  CAS  Google Scholar 

  29. Smith, K.G., Tarlinton, D.M., Doody, G.M., Hibbs, M.L. & Fearon, D.T. Inhibition of the B cell by CD22: a requirement for Lyn. J. Exp. Med. 187, 807–811 (1998).

    Article  CAS  Google Scholar 

  30. Cornall, R.J. et al. Polygenic autoimmune traits: Lyn, CD22, and SHP-1 are limiting elements of a biochemical pathway regulating BCR signaling and selection. Immunity 8, 497–508 (1998).

    Article  CAS  Google Scholar 

  31. Trowbridge, I.S. & Thomas, M.L. CD45: an emerging role as a protein tyrosine phosphatase required for lymphocyte activation and development. Annu. Rev. Immunol. 12, 85–116 (1994).

    Article  CAS  Google Scholar 

  32. Shrivastava, P., Katagiri, T., Ogimoto, M., Mizuno, K. & Yakura, H. Dynamic regulation of Src-family kinases by CD45 in B cells. Blood 103, 1425–1432 (2004).

    Article  CAS  Google Scholar 

  33. Odom, S. et al. Negative regulation of immunoglobulin E-dependent allergic responses by Lyn kinase. J. Exp. Med. 199, 1491–1502 (2004).

    Article  CAS  Google Scholar 

  34. Datta, S.R. et al. Akt phosphorylation of BAD couples survival signals to the cell-intrinsic death machinery. Cell 91, 231–241 (1997).

    Article  CAS  Google Scholar 

  35. Han, S. et al. Cellular interaction in germinal centers. Roles of CD40 ligand and B7-2 in established germinal centers. J. Immunol. 155, 556–567 (1995).

    CAS  PubMed  Google Scholar 

  36. Ghia, P. et al. Unbalanced expression of bcl-2 family proteins in follicular lymphoma: contribution of CD40 signaling in promoting survival. Blood 91, 244–251 (1998).

    CAS  PubMed  Google Scholar 

  37. Tuscano, J.M. et al. Bcl-x rather than Bcl-2 mediates CD40-dependent centrocyte survival in the germinal center. Blood 88, 1359–1364 (1996).

    CAS  PubMed  Google Scholar 

  38. Basso, K. et al. Tracking CD40 signaling during germinal center development. Blood 104, 4088–4096 (2004).

    Article  CAS  Google Scholar 

  39. Foy, T.M. et al. gp39–CD40 interactions are essential for germinal center formation and the development of B cell memory. J. Exp. Med. 180, 157–163 (1994).

    Article  CAS  Google Scholar 

  40. Ridderstad, A., Nossal, G.J. & Tarlinton, D.M. The xid mutation diminishes memory B cell generation but does not affect somatic hypermutation and selection. J. Immunol. 157, 3357–3365 (1996).

    CAS  PubMed  Google Scholar 

  41. Kim, N., Martin, T.E., Simon, M.C. & Storb, U. The transcription factor Spi-B is not required for somatic hypermutation. Mol. Immunol. 39, 577–583 (2003).

    Article  CAS  Google Scholar 

  42. Wang, J.H. et al. Aiolos regulates B cell activation and maturation to effector state. Immunity 9, 543–553 (1998).

    Article  CAS  Google Scholar 

  43. Cortes, M. & Georgopoulos, K. Aiolos is required for the generation of high-affinity bone marrow plasma cells responsible for long-term immunity. J. Exp. Med. 199, 209–219 (2004).

    Article  CAS  Google Scholar 

  44. Smith, K.G., Light, A., Nossal, G.J. & Tarlinton, D.M. The extent of affinity maturation differs between the memory and antibody-forming cell compartments in the primary immune response. EMBO J. 16, 2996–3006 (1997).

    Article  CAS  Google Scholar 

  45. Ng, L.G. et al. B cell-activating factor belonging to the TNF family (BAFF)-R is the principal BAFF receptor facilitating BAFF costimulation of circulating T and B cells. J. Immunol. 173, 807–817 (2004).

    Article  CAS  Google Scholar 

  46. McHeyzer-Williams, M.G., Nossal, G.J. & Lalor, P.A. Molecular characterization of single memory B cells. Nature 350, 502–505 (1991).

    Article  CAS  Google Scholar 

  47. Janas, M.L., Hodgkin, P., Hibbs, M. & Tarlinton, D. Genetic evidence for Lyn as a negative regulator of IL-4 signaling. J. Immunol. 163, 4192–4198 (1999).

    CAS  PubMed  Google Scholar 

  48. Hasbold, J. et al. Quantitative analysis of lymphocyte differentiation and proliferation in vitro using carboxyfluorescein diacetate succinimidyl ester. Immunol. Cell Biol. 77, 516–522 (1999).

    Article  CAS  Google Scholar 

  49. Lalor, P.A., Nossal, G.J., Sanderson, R.D. & McHeyzer-Williams, M.G. Functional and molecular characterization of single, (4-hydroxy-3-nitrophenyl)acetyl (NP)-specific, IgG1+ B cells from antibody-secreting and memory B cell pathways in the C57BL/6 immune response to NP. Eur. J. Immunol. 22, 3001–3011 (1992).

    Article  CAS  Google Scholar 

  50. Pulendran, B., Karvelas, M. & Nossal, G.J. A form of immunologic tolerance through impairment of germinal center development. Proc. Natl. Acad. Sci. USA 91, 2639–2643 (1994).

    Article  CAS  Google Scholar 

  51. Nossal, G.J., Karvelas, M. & Pulendran, B. Soluble antigen profoundly reduces memory B-cell numbers even when given after challenge immunization. Proc. Natl. Acad. Sci. USA 90, 3088–3092 (1993).

    Article  CAS  Google Scholar 

  52. Huang, D.C., Cory, S. & Strasser, A. Bcl-2, Bcl-xL and adenovirus protein E1B19kD are functionally equivalent in their ability to inhibit cell death. Oncogene 14, 405–414 (1997).

    Article  CAS  Google Scholar 

  53. Kinsella, T.M. & Nolan, G.P. Episomal vectors rapidly and stably produce high-titer recombinant retrovirus. Hum. Gene Ther. 7, 1405–1413 (1996).

    Article  CAS  Google Scholar 

  54. Chen, L. et al. Differential targeting of prosurvival Bcl-2 proteins by their BH3-only ligands allows complementary apoptotic function. Mol. Cell 17, 393–403 (2005).

    Article  CAS  Google Scholar 

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Acknowledgements

We thank V. Tybulewicz (National Institute for Medical Research, Mill Hill, UK) for Cd45−/− mice17; K. Rajewsky (Center for Blood Research, Harvard Medical School, Cambridge, Massachusetts) for Cd19−/− mice26 and μMt mice23; D. Huang, G. Belz, J. Silke, L. O'Reilly (The Walter and Eliza Hall Institute of Medical Research, Melbourne, Australia) and F. Mackay (Garvan Institute of Medical Research, Sydney, Australia) for reagents and advice; and B. Mckenzie and S. Holroyd for technical assistance. Supported by the National Health and Medical; Research Council Australia and the Cancer Council of Victoria.

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  1. Yuekang Xu

    Present address: Peter MacCallum Cancer Centre, East Melbourne, Victoria, 3002, Australia

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  1. The Walter and Eliza Hall Institute of Medical Research, Parkville, 3050, Victoria, Australia

    Nicholas D Huntington, Yuekang Xu, Hamsa Puthalakath, Amanda Light, Simon N Willis, Andreas Strasser & David M Tarlinton

  2. Department of Medical Biology, The University of Melbourne, Parkville, 3010, Victoria, Australia

    Nicholas D Huntington

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Supplementary information

Supplementary Fig. 1

General characteristics of exon 12 Cd45−/− B cells. (PDF 171 kb)

Supplementary Fig. 2

CD45-null BCR signaling. (PDF 107 kb)

Supplementary Fig. 3

Enforced expression of Bcl-xL does not alter CD45-null B cell anti-NP response at day 7. (PDF 113 kb)

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Huntington, N., Xu, Y., Puthalakath, H. et al. CD45 links the B cell receptor with cell survival and is required for the persistence of germinal centers. Nat Immunol 7, 190–198 (2006). https://doi.org/10.1038/ni1292

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