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. 1996 Sep 1;184(3):831–838. doi: 10.1084/jem.184.3.831

Altered antigen receptor signaling and impaired Fas-mediated apoptosis of B cells in Lyn-deficient mice

PMCID: PMC2192791  PMID: 9064343

Abstract

Mice deficient in the src related protein tyrosine kinase, Lyn, exhibit splenomegaly and accumulate lymphoblast-like and plasma cells in spleen as they age, resulting in elevated levels of serum IgM (10-20-fold of control) and glomerulonephritis due to the presence of immune complexes containing auto-reactive antibodies. It remains unclear, however, how antibody-producing cells are accumulated in the lymphoid tissues of Lyn- /- mice. To elucidate the role of Lyn in B cell function, we have studied the proliferative responses to various stimuli and Fas-mediated apoptosis in B cells from young Lyn-/- mice which do not yet show apparent abnormality such as splenomegaly. Compared with control B cells, Lyn-/- B cells were hyper responsive to anti-IgM-induced proliferation and defective in Fc gamma RIIB-mediated suppression of B cell antigen receptor (BCR) signaling, indicating that Lyn is involved in the negative regulation of BCR signaling. In addition, the BCR- mediated signal in Lyn-/- B cells, unlike that in control B cells, failed to act in synergy with either CD40- or IL-4 receptor-triggered signal in inducing a strong proliferative response, suggesting that the BCR signaling pathway in Lyn-/- B cells is altered from that in control B cells. Furthermore, Lyn-/- B cells were found to be impaired in the induction of Fas expression after CD40 ligation and exhibited a reduced susceptibility to Fas-mediated apoptosis. Moreover, BCR cross-linking in Lyn-/- B cells suppressed Fas expression induced by costimulation with CD40 ligand and IL-4. Collectively, these results suggest that the accumulation of lymphoblast-like and plasma cells in Lyn-/- mice may be caused in part, by the accelerated activation of B cells in the absence of Lyn, as well as the impaired Fas-mediated apoptosis after the activation.

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Selected References

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  1. Aoki Y., Kim Y. T., Stillwell R., Kim T. J., Pillai S. The SH2 domains of Src family kinases associate with Syk. J Biol Chem. 1995 Jun 30;270(26):15658–15663. doi: 10.1074/jbc.270.26.15658. [DOI] [PubMed] [Google Scholar]
  2. Banchereau J., de Paoli P., Vallé A., Garcia E., Rousset F. Long-term human B cell lines dependent on interleukin-4 and antibody to CD40. Science. 1991 Jan 4;251(4989):70–72. doi: 10.1126/science.1702555. [DOI] [PubMed] [Google Scholar]
  3. Bolen J. B. Protein tyrosine kinases in the initiation of antigen receptor signaling. Curr Opin Immunol. 1995 Jun;7(3):306–311. doi: 10.1016/0952-7915(95)80103-0. [DOI] [PubMed] [Google Scholar]
  4. Brown V. K., Ogle E. W., Burkhardt A. L., Rowley R. B., Bolen J. B., Justement L. B. Multiple components of the B cell antigen receptor complex associate with the protein tyrosine phosphatase, CD45. J Biol Chem. 1994 Jun 24;269(25):17238–17244. [PubMed] [Google Scholar]
  5. Burkhardt A. L., Brunswick M., Bolen J. B., Mond J. J. Anti-immunoglobulin stimulation of B lymphocytes activates src-related protein-tyrosine kinases. Proc Natl Acad Sci U S A. 1991 Aug 15;88(16):7410–7414. doi: 10.1073/pnas.88.16.7410. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Bustelo X. R., Barbacid M. Tyrosine phosphorylation of the vav proto-oncogene product in activated B cells. Science. 1992 May 22;256(5060):1196–1199. doi: 10.1126/science.256.5060.1196. [DOI] [PubMed] [Google Scholar]
  7. Bustelo X. R., Ledbetter J. A., Barbacid M. Product of vav proto-oncogene defines a new class of tyrosine protein kinase substrates. Nature. 1992 Mar 5;356(6364):68–71. doi: 10.1038/356068a0. [DOI] [PubMed] [Google Scholar]
  8. Campbell M. A., Sefton B. M. Association between B-lymphocyte membrane immunoglobulin and multiple members of the Src family of protein tyrosine kinases. Mol Cell Biol. 1992 May;12(5):2315–2321. doi: 10.1128/mcb.12.5.2315. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Campbell M. A., Sefton B. M. Protein tyrosine phosphorylation is induced in murine B lymphocytes in response to stimulation with anti-immunoglobulin. EMBO J. 1990 Jul;9(7):2125–2131. doi: 10.1002/j.1460-2075.1990.tb07381.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Coggeshall K. M., McHugh J. C., Altman A. Predominant expression and activation-induced tyrosine phosphorylation of phospholipase C-gamma 2 in B lymphocytes. Proc Natl Acad Sci U S A. 1992 Jun 15;89(12):5660–5664. doi: 10.1073/pnas.89.12.5660. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Cyster J. G., Goodnow C. C. Protein tyrosine phosphatase 1C negatively regulates antigen receptor signaling in B lymphocytes and determines thresholds for negative selection. Immunity. 1995 Jan;2(1):13–24. doi: 10.1016/1074-7613(95)90075-6. [DOI] [PubMed] [Google Scholar]
  12. D'Ambrosio D., Hippen K. L., Minskoff S. A., Mellman I., Pani G., Siminovitch K. A., Cambier J. C. Recruitment and activation of PTP1C in negative regulation of antigen receptor signaling by Fc gamma RIIB1. Science. 1995 Apr 14;268(5208):293–297. doi: 10.1126/science.7716523. [DOI] [PubMed] [Google Scholar]
  13. Davidson W. F., Morse H. C., 3rd, Sharrow S. O., Chused T. M. Phenotypic and functional effects of the motheaten gene on murine B and T lymphocytes. J Immunol. 1979 Mar;122(3):884–891. [PubMed] [Google Scholar]
  14. Faris M., Gaskin F., Parsons J. T., Fu S. M. CD40 signaling pathway: anti-CD40 monoclonal antibody induces rapid dephosphorylation and phosphorylation of tyrosine-phosphorylated proteins including protein tyrosine kinase Lyn, Fyn, and Syk and the appearance of a 28-kD tyrosine phosphorylated protein. J Exp Med. 1994 Jun 1;179(6):1923–1931. doi: 10.1084/jem.179.6.1923. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Fridman W. H., Bonnerot C., Daeron M., Amigorena S., Teillaud J. L., Sautes C. Structural bases of Fc gamma receptor functions. Immunol Rev. 1992 Feb;125:49–76. doi: 10.1111/j.1600-065x.1992.tb00625.x. [DOI] [PubMed] [Google Scholar]
  16. Garrone P., Neidhardt E. M., Garcia E., Galibert L., van Kooten C., Banchereau J. Fas ligation induces apoptosis of CD40-activated human B lymphocytes. J Exp Med. 1995 Nov 1;182(5):1265–1273. doi: 10.1084/jem.182.5.1265. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Gold M. R., Chan V. W., Turck C. W., DeFranco A. L. Membrane Ig cross-linking regulates phosphatidylinositol 3-kinase in B lymphocytes. J Immunol. 1992 Apr 1;148(7):2012–2022. [PubMed] [Google Scholar]
  18. Gold M. R., Crowley M. T., Martin G. A., McCormick F., DeFranco A. L. Targets of B lymphocyte antigen receptor signal transduction include the p21ras GTPase-activating protein (GAP) and two GAP-associated proteins. J Immunol. 1993 Jan 15;150(2):377–386. [PubMed] [Google Scholar]
  19. Gold M. R., Law D. A., DeFranco A. L. Stimulation of protein tyrosine phosphorylation by the B-lymphocyte antigen receptor. Nature. 1990 Jun 28;345(6278):810–813. doi: 10.1038/345810a0. [DOI] [PubMed] [Google Scholar]
  20. Gold M. R., Matsuuchi L., Kelly R. B., DeFranco A. L. Tyrosine phosphorylation of components of the B-cell antigen receptors following receptor crosslinking. Proc Natl Acad Sci U S A. 1991 Apr 15;88(8):3436–3440. doi: 10.1073/pnas.88.8.3436. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Hibbs M. L., Tarlinton D. M., Armes J., Grail D., Hodgson G., Maglitto R., Stacker S. A., Dunn A. R. Multiple defects in the immune system of Lyn-deficient mice, culminating in autoimmune disease. Cell. 1995 Oct 20;83(2):301–311. doi: 10.1016/0092-8674(95)90171-x. [DOI] [PubMed] [Google Scholar]
  22. Hutchcroft J. E., Harrison M. L., Geahlen R. L. Association of the 72-kDa protein-tyrosine kinase PTK72 with the B cell antigen receptor. J Biol Chem. 1992 Apr 25;267(12):8613–8619. [PubMed] [Google Scholar]
  23. Justement L. B., Campbell K. S., Chien N. C., Cambier J. C. Regulation of B cell antigen receptor signal transduction and phosphorylation by CD45. Science. 1991 Jun 28;252(5014):1839–1842. doi: 10.1126/science.1648262. [DOI] [PubMed] [Google Scholar]
  24. Kehry M. R. CD40-mediated signaling in B cells. Balancing cell survival, growth, and death. J Immunol. 1996 Apr 1;156(7):2345–2348. [PubMed] [Google Scholar]
  25. Kishihara K., Penninger J., Wallace V. A., Kündig T. M., Kawai K., Wakeham A., Timms E., Pfeffer K., Ohashi P. S., Thomas M. L. Normal B lymphocyte development but impaired T cell maturation in CD45-exon6 protein tyrosine phosphatase-deficient mice. Cell. 1993 Jul 16;74(1):143–156. doi: 10.1016/0092-8674(93)90302-7. [DOI] [PubMed] [Google Scholar]
  26. Kozlowski M., Mlinaric-Rascan I., Feng G. S., Shen R., Pawson T., Siminovitch K. A. Expression and catalytic activity of the tyrosine phosphatase PTP1C is severely impaired in motheaten and viable motheaten mice. J Exp Med. 1993 Dec 1;178(6):2157–2163. doi: 10.1084/jem.178.6.2157. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Lane P., Brocker T., Hubele S., Padovan E., Lanzavecchia A., McConnell F. Soluble CD40 ligand can replace the normal T cell-derived CD40 ligand signal to B cells in T cell-dependent activation. J Exp Med. 1993 Apr 1;177(4):1209–1213. doi: 10.1084/jem.177.4.1209. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Li Z. H., Mahajan S., Prendergast M. M., Fargnoli J., Zhu X., Klages S., Adam D., Schieven G. L., Blake J., Bolen J. B. Cross-linking of surface immunoglobulin activates src-related tyrosine kinases in WEHI 231 cells. Biochem Biophys Res Commun. 1992 Sep 30;187(3):1536–1544. doi: 10.1016/0006-291x(92)90477-3. [DOI] [PubMed] [Google Scholar]
  29. Liu Y. J., Joshua D. E., Williams G. T., Smith C. A., Gordon J., MacLennan I. C. Mechanism of antigen-driven selection in germinal centres. Nature. 1989 Dec 21;342(6252):929–931. doi: 10.1038/342929a0. [DOI] [PubMed] [Google Scholar]
  30. Margolis B., Hu P., Katzav S., Li W., Oliver J. M., Ullrich A., Weiss A., Schlessinger J. Tyrosine phosphorylation of vav proto-oncogene product containing SH2 domain and transcription factor motifs. Nature. 1992 Mar 5;356(6364):71–74. doi: 10.1038/356071a0. [DOI] [PubMed] [Google Scholar]
  31. Nagata S., Suda T. Fas and Fas ligand: lpr and gld mutations. Immunol Today. 1995 Jan;16(1):39–43. doi: 10.1016/0167-5699(95)80069-7. [DOI] [PubMed] [Google Scholar]
  32. Nishizumi H., Taniuchi I., Yamanashi Y., Kitamura D., Ilic D., Mori S., Watanabe T., Yamamoto T. Impaired proliferation of peripheral B cells and indication of autoimmune disease in lyn-deficient mice. Immunity. 1995 Nov;3(5):549–560. doi: 10.1016/1074-7613(95)90126-4. [DOI] [PubMed] [Google Scholar]
  33. Okumura M., Thomas M. L. Regulation of immune function by protein tyrosine phosphatases. Curr Opin Immunol. 1995 Jun;7(3):312–319. doi: 10.1016/0952-7915(95)80104-9. [DOI] [PubMed] [Google Scholar]
  34. Pani G., Kozlowski M., Cambier J. C., Mills G. B., Siminovitch K. A. Identification of the tyrosine phosphatase PTP1C as a B cell antigen receptor-associated protein involved in the regulation of B cell signaling. J Exp Med. 1995 Jun 1;181(6):2077–2084. doi: 10.1084/jem.181.6.2077. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Pesando J. M., Bouchard L. S., McMaster B. E. CD19 is functionally and physically associated with surface immunoglobulin. J Exp Med. 1989 Dec 1;170(6):2159–2164. doi: 10.1084/jem.170.6.2159. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Pezzutto A., Dörken B., Rabinovitch P. S., Ledbetter J. A., Moldenhauer G., Clark E. A. CD19 monoclonal antibody HD37 inhibits anti-immunoglobulin-induced B cell activation and proliferation. J Immunol. 1987 May 1;138(9):2793–2799. [PubMed] [Google Scholar]
  37. Phillips N. E., Parker D. C. Fc-dependent inhibition of mouse B cell activation by whole anti-mu antibodies. J Immunol. 1983 Feb;130(2):602–606. [PubMed] [Google Scholar]
  38. Ren C. L., Morio T., Fu S. M., Geha R. S. Signal transduction via CD40 involves activation of lyn kinase and phosphatidylinositol-3-kinase, and phosphorylation of phospholipase C gamma 2. J Exp Med. 1994 Feb 1;179(2):673–680. doi: 10.1084/jem.179.2.673. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Rothstein T. L., Wang J. K., Panka D. J., Foote L. C., Wang Z., Stanger B., Cui H., Ju S. T., Marshak-Rothstein A. Protection against Fas-dependent Th1-mediated apoptosis by antigen receptor engagement in B cells. Nature. 1995 Mar 9;374(6518):163–165. doi: 10.1038/374163a0. [DOI] [PubMed] [Google Scholar]
  40. Saxton T. M., van Oostveen I., Bowtell D., Aebersold R., Gold M. R. B cell antigen receptor cross-linking induces phosphorylation of the p21ras oncoprotein activators SHC and mSOS1 as well as assembly of complexes containing SHC, GRB-2, mSOS1, and a 145-kDa tyrosine-phosphorylated protein. J Immunol. 1994 Jul 15;153(2):623–636. [PubMed] [Google Scholar]
  41. Schattner E. J., Elkon K. B., Yoo D. H., Tumang J., Krammer P. H., Crow M. K., Friedman S. M. CD40 ligation induces Apo-1/Fas expression on human B lymphocytes and facilitates apoptosis through the Apo-1/Fas pathway. J Exp Med. 1995 Nov 1;182(5):1557–1565. doi: 10.1084/jem.182.5.1557. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Scheuermann R. H., Racila E., Tucker T., Yefenof E., Street N. E., Vitetta E. S., Picker L. J., Uhr J. W. Lyn tyrosine kinase signals cell cycle arrest but not apoptosis in B-lineage lymphoma cells. Proc Natl Acad Sci U S A. 1994 Apr 26;91(9):4048–4052. doi: 10.1073/pnas.91.9.4048. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Shultz L. D., Schweitzer P. A., Rajan T. V., Yi T., Ihle J. N., Matthews R. J., Thomas M. L., Beier D. R. Mutations at the murine motheaten locus are within the hematopoietic cell protein-tyrosine phosphatase (Hcph) gene. Cell. 1993 Jul 2;73(7):1445–1454. doi: 10.1016/0092-8674(93)90369-2. [DOI] [PubMed] [Google Scholar]
  44. Sidman C. L., Shultz L. D., Hardy R. R., Hayakawa K., Herzenberg L. A. Production of immunoglobulin isotypes by Ly-1+ B cells in viable motheaten and normal mice. Science. 1986 Jun 13;232(4756):1423–1425. doi: 10.1126/science.3487115. [DOI] [PubMed] [Google Scholar]
  45. Sidorenko S. P., Law C. L., Chandran K. A., Clark E. A. Human spleen tyrosine kinase p72Syk associates with the Src-family kinase p53/56Lyn and a 120-kDa phosphoprotein. Proc Natl Acad Sci U S A. 1995 Jan 17;92(2):359–363. doi: 10.1073/pnas.92.2.359. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Takai T., Ono M., Hikida M., Ohmori H., Ravetch J. V. Augmented humoral and anaphylactic responses in Fc gamma RII-deficient mice. Nature. 1996 Jan 25;379(6563):346–349. doi: 10.1038/379346a0. [DOI] [PubMed] [Google Scholar]
  47. Takata M., Sabe H., Hata A., Inazu T., Homma Y., Nukada T., Yamamura H., Kurosaki T. Tyrosine kinases Lyn and Syk regulate B cell receptor-coupled Ca2+ mobilization through distinct pathways. EMBO J. 1994 Mar 15;13(6):1341–1349. doi: 10.1002/j.1460-2075.1994.tb06387.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Tezuka T., Umemori H., Fusaki N., Yagi T., Takata M., Kurosaki T., Yamamoto T. Physical and functional association of the cbl protooncogen product with an src-family protein tyrosine kinase, p53/56lyn, in the B cell antigen receptor-mediated signaling. J Exp Med. 1996 Feb 1;183(2):675–680. doi: 10.1084/jem.183.2.675. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Tsui H. W., Siminovitch K. A., de Souza L., Tsui F. W. Motheaten and viable motheaten mice have mutations in the haematopoietic cell phosphatase gene. Nat Genet. 1993 Jun;4(2):124–129. doi: 10.1038/ng0693-124. [DOI] [PubMed] [Google Scholar]
  50. Umemori H., Wanaka A., Kato H., Takeuchi M., Tohyama M., Yamamoto T. Specific expressions of Fyn and Lyn, lymphocyte antigen receptor-associated tyrosine kinases, in the central nervous system. Brain Res Mol Brain Res. 1992 Dec;16(3-4):303–310. doi: 10.1016/0169-328x(92)90239-8. [DOI] [PubMed] [Google Scholar]
  51. Wang J., Taniuchi I., Maekawa Y., Howard M., Cooper M. D., Watanabe T. Expression and function of Fas antigen on activated murine B cells. Eur J Immunol. 1996 Jan;26(1):92–96. doi: 10.1002/eji.1830260114. [DOI] [PubMed] [Google Scholar]
  52. Yamada T., Taniguchi T., Yang C., Yasue S., Saito H., Yamamura H. Association with B-cell-antigen receptor with protein-tyrosine kinase p72syk and activation by engagement of membrane IgM. Eur J Biochem. 1993 Apr 1;213(1):455–459. doi: 10.1111/j.1432-1033.1993.tb17781.x. [DOI] [PubMed] [Google Scholar]
  53. Yamanashi Y., Fukui Y., Wongsasant B., Kinoshita Y., Ichimori Y., Toyoshima K., Yamamoto T. Activation of Src-like protein-tyrosine kinase Lyn and its association with phosphatidylinositol 3-kinase upon B-cell antigen receptor-mediated signaling. Proc Natl Acad Sci U S A. 1992 Feb 1;89(3):1118–1122. doi: 10.1073/pnas.89.3.1118. [DOI] [PMC free article] [PubMed] [Google Scholar]
  54. Yamanashi Y., Kakiuchi T., Mizuguchi J., Yamamoto T., Toyoshima K. Association of B cell antigen receptor with protein tyrosine kinase Lyn. Science. 1991 Jan 11;251(4990):192–194. doi: 10.1126/science.1702903. [DOI] [PubMed] [Google Scholar]
  55. Yamanashi Y., Mori S., Yoshida M., Kishimoto T., Inoue K., Yamamoto T., Toyoshima K. Selective expression of a protein-tyrosine kinase, p56lyn, in hematopoietic cells and association with production of human T-cell lymphotropic virus type I. Proc Natl Acad Sci U S A. 1989 Sep;86(17):6538–6542. doi: 10.1073/pnas.86.17.6538. [DOI] [PMC free article] [PubMed] [Google Scholar]
  56. Yamanashi Y., Okada M., Semba T., Yamori T., Umemori H., Tsunasawa S., Toyoshima K., Kitamura D., Watanabe T., Yamamoto T. Identification of HS1 protein as a major substrate of protein-tyrosine kinase(s) upon B-cell antigen receptor-mediated signaling. Proc Natl Acad Sci U S A. 1993 Apr 15;90(8):3631–3635. doi: 10.1073/pnas.90.8.3631. [DOI] [PMC free article] [PubMed] [Google Scholar]
  57. van Noesel C. J., Lankester A. C., van Schijndel G. M., van Lier R. A. The CR2/CD19 complex on human B cells contains the src-family kinase Lyn. Int Immunol. 1993 Jul;5(7):699–705. doi: 10.1093/intimm/5.7.699. [DOI] [PubMed] [Google Scholar]

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