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. 1996 Sep 2;15(17):4682–4690.

Inhibition of NF-kappaB/Rel induces apoptosis of murine B cells.

M Wu 1, H Lee 1, R E Bellas 1, S L Schauer 1, M Arsura 1, D Katz 1, M J FitzGerald 1, T L Rothstein 1, D H Sherr 1, G E Sonenshein 1
PMCID: PMC452200  PMID: 8887559

Abstract

Apoptosis of the WEHI 231 immature B cell lymphoma line following membrane interaction with an antibody against the surface IgM chains (anti-IgM) is preceded by dramatic changes in Nuclear Factor-kappaB (NF-kappaB)/ Rel binding activities. An early transient increase in NF-kappaB/Rel binding is followed by a significant decrease in intensity below basal levels. Here we have explored the role of these changes in Rel-related factors in B cell apoptosis. Treatment of WEH1 231 cells with N-tosyl-L-phenylalanine chloromethyl ketone (TPCK), a protease inhibitor which prevents degradation of the inhibitor of NF-kappaB (IkappaB)-alpha, or with low doses of pyrrolidinedithiocarbamate (PDTC) selectively inhibited NF-kappaB/Rel factor binding and induced apoptosis. Bcl-XL expression protected WEHI 231 cells from apoptosis induced by these agents. Microinjection of WEHI 231 cells with either IkappaB-alpha-GST protein or a c-Rel affinity-purified antibody induced apoptosis. Ectopic c-Rel expression ablated apoptosis induced by TPCK or anti-IgM. Treatment of BALENLM 17 and A20 B lymphoma cells or normal murine splenic B lymphocytes with either TPCK or PDTC also resulted in apoptosis. These findings indicate that the drop in NF-kappaB/Rel binding following anti-IgM treatment activates apoptosis of WEHI 231 cells; furthermore, they implicate the NF-kappaB/Rel family in control of apoptosis of normal and transformed B cells.

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Selected References

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

  1. Abbadie C., Kabrun N., Bouali F., Smardova J., Stéhelin D., Vandenbunder B., Enrietto P. J. High levels of c-rel expression are associated with programmed cell death in the developing avian embryo and in bone marrow cells in vitro. Cell. 1993 Dec 3;75(5):899–912. doi: 10.1016/0092-8674(93)90534-w. [DOI] [PubMed] [Google Scholar]
  2. Arsura M., Deshpande A., Hann S. R., Sonenshein G. E. Variant Max protein, derived by alternative splicing, associates with c-Myc in vivo and inhibits transactivation. Mol Cell Biol. 1995 Dec;15(12):6702–6709. doi: 10.1128/mcb.15.12.6702. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Baeuerle P. A., Baltimore D. I kappa B: a specific inhibitor of the NF-kappa B transcription factor. Science. 1988 Oct 28;242(4878):540–546. doi: 10.1126/science.3140380. [DOI] [PubMed] [Google Scholar]
  4. Baeuerle P. A. The inducible transcription activator NF-kappa B: regulation by distinct protein subunits. Biochim Biophys Acta. 1991 Apr 16;1072(1):63–80. doi: 10.1016/0304-419x(91)90007-8. [DOI] [PubMed] [Google Scholar]
  5. Beg A. A., Sha W. C., Bronson R. T., Ghosh S., Baltimore D. Embryonic lethality and liver degeneration in mice lacking the RelA component of NF-kappa B. Nature. 1995 Jul 13;376(6536):167–170. doi: 10.1038/376167a0. [DOI] [PubMed] [Google Scholar]
  6. 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]
  7. Bessho R., Matsubara K., Kubota M., Kuwakado K., Hirota H., Wakazono Y., Lin Y. W., Okuda A., Kawai M., Nishikomori R. Pyrrolidine dithiocarbamate, a potent inhibitor of nuclear factor kappa B (NF-kappa B) activation, prevents apoptosis in human promyelocytic leukemia HL-60 cells and thymocytes. Biochem Pharmacol. 1994 Nov 16;48(10):1883–1889. doi: 10.1016/0006-2952(94)90586-x. [DOI] [PubMed] [Google Scholar]
  8. Boyd A. W., Schrader J. W. The regulation of growth and differentiation of a murine B cell lymphoma. II. The inhibition of WEHI 231 by anti-immunoglobulin antibodies. J Immunol. 1981 Jun;126(6):2466–2469. [PubMed] [Google Scholar]
  9. Brown K., Gerstberger S., Carlson L., Franzoso G., Siebenlist U. Control of I kappa B-alpha proteolysis by site-specific, signal-induced phosphorylation. Science. 1995 Mar 10;267(5203):1485–1488. doi: 10.1126/science.7878466. [DOI] [PubMed] [Google Scholar]
  10. Duyao M. P., Buckler A. J., Sonenshein G. E. Interaction of an NF-kappa B-like factor with a site upstream of the c-myc promoter. Proc Natl Acad Sci U S A. 1990 Jun;87(12):4727–4731. doi: 10.1073/pnas.87.12.4727. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Duyao M. P., Kessler D. J., Spicer D. B., Bartholomew C., Cleveland J. L., Siekevitz M., Sonenshein G. E. Transactivation of the c-myc promoter by human T cell leukemia virus type 1 tax is mediated by NF kappa B. J Biol Chem. 1992 Aug 15;267(23):16288–16291. [PubMed] [Google Scholar]
  12. Finco T. S., Beg A. A., Baldwin A. S., Jr Inducible phosphorylation of I kappa B alpha is not sufficient for its dissociation from NF-kappa B and is inhibited by protease inhibitors. Proc Natl Acad Sci U S A. 1994 Dec 6;91(25):11884–11888. doi: 10.1073/pnas.91.25.11884. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Fischer G., Kent S. C., Joseph L., Green D. R., Scott D. W. Lymphoma models for B cell activation and tolerance. X. Anti-mu-mediated growth arrest and apoptosis of murine B cell lymphomas is prevented by the stabilization of myc. J Exp Med. 1994 Jan 1;179(1):221–228. doi: 10.1084/jem.179.1.221. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Gottschalk A. R., Boise L. H., Thompson C. B., Quintáns J. Identification of immunosuppressant-induced apoptosis in a murine B-cell line and its prevention by bcl-x but not bcl-2. Proc Natl Acad Sci U S A. 1994 Jul 19;91(15):7350–7354. doi: 10.1073/pnas.91.15.7350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Grilli M., Chiu J. J., Lenardo M. J. NF-kappa B and Rel: participants in a multiform transcriptional regulatory system. Int Rev Cytol. 1993;143:1–62. doi: 10.1016/s0074-7696(08)61873-2. [DOI] [PubMed] [Google Scholar]
  16. 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]
  17. Henkel T., Machleidt T., Alkalay I., Krönke M., Ben-Neriah Y., Baeuerle P. A. Rapid proteolysis of I kappa B-alpha is necessary for activation of transcription factor NF-kappa B. Nature. 1993 Sep 9;365(6442):182–185. doi: 10.1038/365182a0. [DOI] [PubMed] [Google Scholar]
  18. Hsu H., Shu H. B., Pan M. G., Goeddel D. V. TRADD-TRAF2 and TRADD-FADD interactions define two distinct TNF receptor 1 signal transduction pathways. Cell. 1996 Jan 26;84(2):299–308. doi: 10.1016/s0092-8674(00)80984-8. [DOI] [PubMed] [Google Scholar]
  19. Hsu H., Xiong J., Goeddel D. V. The TNF receptor 1-associated protein TRADD signals cell death and NF-kappa B activation. Cell. 1995 May 19;81(4):495–504. doi: 10.1016/0092-8674(95)90070-5. [DOI] [PubMed] [Google Scholar]
  20. Kenter A. L., Wuerffel R., Sen R., Jamieson C. E., Merkulov G. V. Switch recombination breakpoints occur at nonrandom positions in the S gamma tandem repeat. J Immunol. 1993 Nov 1;151(9):4718–4731. [PubMed] [Google Scholar]
  21. Kessler D. J., Duyao M. P., Spicer D. B., Sonenshein G. E. NF-kappa B-like factors mediate interleukin 1 induction of c-myc gene transcription in fibroblasts. J Exp Med. 1992 Sep 1;176(3):787–792. doi: 10.1084/jem.176.3.787. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Kim K. J., Kanellopoulos-Langevin C., Merwin R. M., Sachs D. H., Asofsky R. Establishment and characterization of BALB/c lymphoma lines with B cell properties. J Immunol. 1979 Feb;122(2):549–554. [PubMed] [Google Scholar]
  23. La Rosa F. A., Pierce J. W., Sonenshein G. E. Differential regulation of the c-myc oncogene promoter by the NF-kappa B rel family of transcription factors. Mol Cell Biol. 1994 Feb;14(2):1039–1044. doi: 10.1128/mcb.14.2.1039. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Lee H., Arsura M., Wu M., Duyao M., Buckler A. J., Sonenshein G. E. Role of Rel-related factors in control of c-myc gene transcription in receptor-mediated apoptosis of the murine B cell WEHI 231 line. J Exp Med. 1995 Mar 1;181(3):1169–1177. doi: 10.1084/jem.181.3.1169. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Lee H., Wu M., La Rosa F. A., Duyao M. P., Buckler A. J., Sonenshein G. E. Role of the Rel-family of transcription factors in the regulation of c-myc gene transcription and apoptosis of WEHI 231 murine B-cells. Curr Top Microbiol Immunol. 1995;194:247–255. doi: 10.1007/978-3-642-79275-5_29. [DOI] [PubMed] [Google Scholar]
  26. Liou H. C., Sha W. C., Scott M. L., Baltimore D. Sequential induction of NF-kappa B/Rel family proteins during B-cell terminal differentiation. Mol Cell Biol. 1994 Aug;14(8):5349–5359. doi: 10.1128/mcb.14.8.5349. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Liu J. L., Chiles T. C., Sen R. J., Rothstein T. L. Inducible nuclear expression of NF-kappa B in primary B cells stimulated through the surface Ig receptor. J Immunol. 1991 Mar 1;146(5):1685–1691. [PubMed] [Google Scholar]
  28. McCormack J. E., Pepe V. H., Kent R. B., Dean M., Marshak-Rothstein A., Sonenshein G. E. Specific regulation of c-myc oncogene expression in a murine B-cell lymphoma. Proc Natl Acad Sci U S A. 1984 Sep;81(17):5546–5550. doi: 10.1073/pnas.81.17.5546. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Merino R., Grillot D. A., Simonian P. L., Muthukkumar S., Fanslow W. C., Bondada S., Núez G. Modulation of anti-IgM-induced B cell apoptosis by Bcl-xL and CD40 in WEHI-231 cells. Dissociation from cell cycle arrest and dependence on the avidity of the antibody-IgM receptor interaction. J Immunol. 1995 Oct 15;155(8):3830–3838. [PubMed] [Google Scholar]
  30. Miyamoto S., Chiao P. J., Verma I. M. Enhanced I kappa B alpha degradation is responsible for constitutive NF-kappa B activity in mature murine B-cell lines. Mol Cell Biol. 1994 May;14(5):3276–3282. doi: 10.1128/mcb.14.5.3276. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Monroe J. G., Seyfert V. L. Studies of surface immunoglobulin-dependent B cell activation. Immunol Res. 1988;7(2):136–151. doi: 10.1007/BF02918097. [DOI] [PubMed] [Google Scholar]
  32. Neiman P. E., Thomas S. J., Loring G. Induction of apoptosis during normal and neoplastic B-cell development in the bursa of Fabricius. Proc Natl Acad Sci U S A. 1991 Jul 1;88(13):5857–5861. doi: 10.1073/pnas.88.13.5857. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Perez P., Lira S. A., Bravo R. Overexpression of RelA in transgenic mouse thymocytes: specific increase in levels of the inhibitor protein I kappa B alpha. Mol Cell Biol. 1995 Jul;15(7):3523–3530. doi: 10.1128/mcb.15.7.3523. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Rice N. R., Ernst M. K. In vivo control of NF-kappa B activation by I kappa B alpha. EMBO J. 1993 Dec;12(12):4685–4695. doi: 10.1002/j.1460-2075.1993.tb06157.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. 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]
  36. Schauer S. L., Wang Z., Sonenshein G. E., Rothstein T. L. Maintenance of nuclear factor-kappa B/Rel and c-myc expression during CD40 ligand rescue of WEHI 231 early B cells from receptor-mediated apoptosis through modulation of I kappa B proteins. J Immunol. 1996 Jul 1;157(1):81–86. [PubMed] [Google Scholar]
  37. Schenk H., Klein M., Erdbrügger W., Dröge W., Schulze-Osthoff K. Distinct effects of thioredoxin and antioxidants on the activation of transcription factors NF-kappa B and AP-1. Proc Natl Acad Sci U S A. 1994 Mar 1;91(5):1672–1676. doi: 10.1073/pnas.91.5.1672. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Schreck R., Meier B., Männel D. N., Dröge W., Baeuerle P. A. Dithiocarbamates as potent inhibitors of nuclear factor kappa B activation in intact cells. J Exp Med. 1992 May 1;175(5):1181–1194. doi: 10.1084/jem.175.5.1181. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Scott D. W., Livnat D., Pennell C. A., Keng P. Lymphoma models for B cell activation and tolerance. III. Cell cycle dependence for negative signalling of WEHI-231 B lymphoma cells by anti-mu. J Exp Med. 1986 Jul 1;164(1):156–164. doi: 10.1084/jem.164.1.156. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Sen R., Baltimore D. Inducibility of kappa immunoglobulin enhancer-binding protein Nf-kappa B by a posttranslational mechanism. Cell. 1986 Dec 26;47(6):921–928. doi: 10.1016/0092-8674(86)90807-x. [DOI] [PubMed] [Google Scholar]
  41. Sen R., Baltimore D. Multiple nuclear factors interact with the immunoglobulin enhancer sequences. Cell. 1986 Aug 29;46(5):705–716. doi: 10.1016/0092-8674(86)90346-6. [DOI] [PubMed] [Google Scholar]
  42. Smith C. A., Williams G. T., Kingston R., Jenkinson E. J., Owen J. J. Antibodies to CD3/T-cell receptor complex induce death by apoptosis in immature T cells in thymic cultures. Nature. 1989 Jan 12;337(6203):181–184. doi: 10.1038/337181a0. [DOI] [PubMed] [Google Scholar]
  43. Strauss F., Varshavsky A. A protein binds to a satellite DNA repeat at three specific sites that would be brought into mutual proximity by DNA folding in the nucleosome. Cell. 1984 Jul;37(3):889–901. doi: 10.1016/0092-8674(84)90424-0. [DOI] [PubMed] [Google Scholar]
  44. Sun S. C., Ganchi P. A., Ballard D. W., Greene W. C. NF-kappa B controls expression of inhibitor I kappa B alpha: evidence for an inducible autoregulatory pathway. Science. 1993 Mar 26;259(5103):1912–1915. doi: 10.1126/science.8096091. [DOI] [PubMed] [Google Scholar]
  45. Sun X. M., Snowden R. T., Skilleter D. N., Dinsdale D., Ormerod M. G., Cohen G. M. A flow-cytometric method for the separation and quantitation of normal and apoptotic thymocytes. Anal Biochem. 1992 Aug 1;204(2):351–356. doi: 10.1016/0003-2697(92)90251-2. [DOI] [PubMed] [Google Scholar]
  46. Tisch R., Roifman C. M., Hozumi N. Functional differences between immunoglobulins M and D expressed on the surface of an immature B-cell line. Proc Natl Acad Sci U S A. 1988 Sep;85(18):6914–6918. doi: 10.1073/pnas.85.18.6914. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Tsubata T., Wu J., Honjo T. B-cell apoptosis induced by antigen receptor crosslinking is blocked by a T-cell signal through CD40. Nature. 1993 Aug 12;364(6438):645–648. doi: 10.1038/364645a0. [DOI] [PubMed] [Google Scholar]
  48. White D. W., Roy A., Gilmore T. D. The v-Rel oncoprotein blocks apoptosis and proteolysis of I kappa B-alpha in transformed chicken spleen cells. Oncogene. 1995 Mar 2;10(5):857–868. [PubMed] [Google Scholar]

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