Regulation of B Cell Apoptosis

  • Robert F. Ashman
  • David Peckham
  • Laura L. Stunz
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 406)

Abstract

Only 1% of thymocytes have hypodiploid nuclei, an index of apoptosis. Yet there is evidence that 99% of thymocytes die by apoptosis, and are destroyed so quickly by phagocytes they are hardly apparent histologically. So when we see 1% of spleen B (or T) cells have hypodiploid nuclei, it tells us little about the apoptotic turnover of these cells. Despite evidence that germinal center B cells enter apoptosis when activated through their antigen receptor in the absence of costimulation1, and that terminal differentiation also leads to apoptosis, life span estimates for the majority of mature B cells run to several months2,3. So we were surprised to find that small dense B cells from the spleens of specific pathogen-free mice spontaneously undergo apoptosis in vitro 4 so fast that about 75% of small dense B cells have hypodiploid nuclei by 64h. The rate is linear with no lag period to indicate an induction process. Varying fetal calf serum concentration from 1 to 10%, varying the B cell (or T cell) concentration from 0.1 to 10 × 106/ml (to rule out autocrine mechanisms), and purifying the cells (adherence to BSA-coated plastic, exposure to anti-thy 1.2 plus complement, centrifugation through Percoll) had no major effect on the spontaneous apoptosis rate5(and our unpublished data). Neither did inclusion of 10% normal mouse serum, or coculture with other spleen cell types5. Cycloheximide (CHX) and other protein synthesis inhibitors accelerated apoptosis in B and T cells4,5 while blocking apoptosis in thymocytes5,6. We appeared to be observing an apoptosis program “hard-wired” into the B cell, very different from that of the thymocyte, requiring no stimulation and no “death protein” synthesis.

Keywords

Spleen Cell Acridine Orange Apoptosis Rate Dextran Sulfate None None 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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References

  1. 1.
    MacLennan ICM, Liu Y-J, Johnson G. 1992. Maturation and dispersal of B cell clones during T-dependent antibody responses. Immunol Rev 126: 143–161.PubMedCrossRefGoogle Scholar
  2. 2.
    Grabstein KH, Waldschmidt TJ, Finkelman FD, Hess BW, Alpert AR, Boiani NE, Namen AE, Morrissey PJ. 1993. Inhibition of murine B and T lymphopoiesis in vivo by an anti-interleukin 7 monoclonal antibody. J. of Exper. Med. 178: 257–264.CrossRefGoogle Scholar
  3. 3.
    Forster I, and Rajewsky K. 1990. The bulk of the peripheral B-cell pool in mice is stable and not rapidly renewed from the bone marrow. Proc. Natl. Acad. Sci. USA 87 (12): 4781–4789.PubMedCrossRefGoogle Scholar
  4. 4.
    Illera VA, Perandones CE, Stunz LL, Mower DA, Jr., and Ashman RF. 1993. Apoptosis in splenic B lymphocytes: Regulation by protein kinase C and IL-4. J. of Immunol., 151: 2965.Google Scholar
  5. 5.
    Perandones CD, Illera VA, Peckham D, Stunz LL, and Ashman RF. 1993. Regulation of apoptosis in vitro in mature murine spleen T cells. J. of Immunol., 151: 3521.Google Scholar
  6. 6.
    Cohen JJ, and Duke RC. 1984. Clucocorticoid activation of a calcium-dependent endonuclease in thymocyte nuclei leads to cell death. J. Immunol., 132: 38.PubMedGoogle Scholar
  7. 7.
    Fadok VA, Voelker DR, Campbell PA, Cohen JJ, Bratton DL, and Henson PM. 1992. Exposure of phosphatidylserine on the surface of apoptotic lymphocytes triggers specific recognition and removal by macrophages. J. Immunol. 148: 2207–2216.PubMedGoogle Scholar
  8. 8.
    Wetzel GD, Swain SL, Dutton RW, and Kettman JR. 1984. Evidence for two distinct activation states available to B lymphocytes. J. Immunol. 133: 2327.PubMedGoogle Scholar
  9. 9.
    Minchin SA, Leitenberg D, Stunz LL, and Feldbush TL. 1990. Polyclonal activation of rat B cells. J. of Immunol. 145: 2427.Google Scholar
  10. 10.
    Bradbury MG, and Parish CR. 1991. Characterization of lymphocyte receptors for glycosaminoglycans. Immunol. 72: 231.Google Scholar
  11. 11.
    Mower DA, Jr., Peckham DW, Illera VA, Fishbaugh JK, Stunz LL, and Ashman RF. 1994. Decreased membrane phospholipid packing and decreased cell size precede DNA cleavage in mature moust B cell apoptosis. J. Immunol. 152: 4832.PubMedGoogle Scholar
  12. 12.
    Bell DA, and Morrison B. 1991. The spontaneous apoptotic cell death of normal human lymphocytes in vitro: the release of, and immunoproliferative response to, nucleosomes in vitro. Clin. Immunol. Immunopath 60: 13.CrossRefGoogle Scholar
  13. 13.
    Green DR, Mahboubi A, Nishioka W, Oja S, Echeverri F, Shi Y, Glynn J, Yang Y, Ashwell J, and Bissonnette R. 1994. Promotion and inhibition of activation-induced apoptosis in T-cell hybridomas by oncogenes and related signals. Immunol. Rev., 142: 321–342.Google Scholar
  14. 14.
    Shi Y, Bissonnette RP, Glynn JM, Guilbert LJ, Cotter TG, and Green DR. 1992. Role for c-myc in activation-induced apoptotic cell death in T cell hybridomas. Science 257: 212.PubMedCrossRefGoogle Scholar
  15. 15.
    Finkelman FD, Scher I, Mond JJ, Kung JT, and Metcalf ES. 1982. Polyclonal activation of the murine immune system by an antibody to IgD. I. Increase in cell size and DNA synthesis. J. Immunol. 129: 629–637.Google Scholar
  16. 16.
    Finkelman FD, Holmes JM, Dulchanina OL, and Morris SC. 1996. Crosslinking of membrane IgD, in the absence of T cell help, kills mature B cells in vivo. (ms submitted).Google Scholar
  17. 17.
    Phillips NE, and Parker DC. 1984. Cross-linking of B lymphocyte Fcy receptors and membrane immunoglobulin inhibits anti-immunoglobulin-induced blastogenesis. J. Immunol. 6: 1777.Google Scholar
  18. 18.
    O’Garra A, Rigley KP, Holman M, McGlaughlan JB, and Klaus GGB. 1987. B cell stimulatory factor 1 reverses Fc receptor mediated inhibition of B lymphocyte activation. Proc. Natl. Acad. Sci. USA, 84: 6254.Google Scholar
  19. 19.
    D’Ambrosio D, Hippen KL, Minskoff SA, Mellman I, Pani G, Siminovitch KA, and Cambier JC. 1995. Recruitment and activation of PTP IC in negative regulation of antigen receptor signaling by FcyRIII31. Science 268: 293.PubMedCrossRefGoogle Scholar
  20. 20.
    Su X, Zhou T, Wang Z, Yang PA, Jope RS, and Mountz JD. 1995. Defective expression of hematopoietic cell protein tyrosine phosphatase ( HCP) in lymphoid cells blocks fas-mediated apoptosis. Immunity 2: 353–362.PubMedCrossRefGoogle Scholar
  21. 21.
    McDonnell TJ, Nunez G, Platt FM, Hockenbery D, London L, McKearn JP, and Korsmeyer SJ. 1990. Deregulated bcl-2-immunoglobulin transgene expands a resting but responsive immunoglobulin M and D-expressing B-cell population. Molecular and Cell. Biol. 10: 1901–1907.Google Scholar

Copyright information

© Springer Science+Business Media New York 1996

Authors and Affiliations

  • Robert F. Ashman
    • 1
    • 2
  • David Peckham
    • 1
  • Laura L. Stunz
    • 1
  1. 1.Iowa City DVA Medical CenterUniversity of Iowa College of MedicineIowa CityUSA
  2. 2.Department of Medicine and Department of MicrobiologyUniversity of Iowa College of MedicineIowa CityUSA

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