International Journal of Hematology

, Volume 76, Issue 1, pp 44–49 | Cite as

Role of Ca2+ in the Intracellular Signaling Pathway of Anti-IgM—Induced Apoptosis in the Human B-Cell Line, MBC-1, Established from Burkitt Lymphoma

  • Masamune Higashigawa
  • Yoshihiro Komada
Review Article


The role of Ca2+ in the intracellular signal transduction process that causes antibody-induced apoptotic cell death in B-cells is not completely understood. We previously established a B-cell line (MBC-1) from a patient with Burkitt lymphoma at the leukemic stage that demonstrated the typical morphology and internucleosomal DNA fragmentation of apoptosis when treated with anti-immunoglobulin (Ig)M antibody. This antibody-induced cell death was partially inhibited by pretreatment with ethyleneglycol-bis-tetraacetic acid (EGTA) and actinomycin-D. FK506, an immunosupressive agent and calcineurin inhibitor, also partially rescued the anti-IgM antibody—induced death of MBC-1 cells. These results show that the calcium signaling pathway, which leads to a change in gene expression, plays an important role in anti-IgM—induced apoptosis in MBC-1 cells. Flow cytometric measurement of the cytosolic free Ca2+ concentration ([Ca2+]i) showed that nontoxic concentrations of 4-bromo-calcium ionophore A23187 (Ca2+ IP) increased [Ca2+]i more than did the anti-IgM antibody. A brief Ca2+ spike was observed on anti-IgM antibody treatment, but a gradual increase and decrease were observed when the cells were treated with Ca2+ IP at a nontoxic concentration of 1 μg/mL. These findings suggest that interpretations differ for the 2 patterns of calcium signaling and that the brief spiked elevation of Ca2+ produces distinct biological and cellular responses compared to the gradual increase and decrease of [Ca2+]i. Our results support the hypothesis that Ca2+ plays a significant role as a multifunctional second messenger providing specific information to the nucleus in anti-IgM antibody—induced apoptosis in MBC-1 cells.

Key words

B-cell Apoptosis Anti-IgM antibody Ca2+ signaling Calcineurin FK506 


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  1. 1.
    Pratt RB, Greene RM. Inhibition of palatal epithelial cell death by altered protein synthesis.Devel Biol. 1976;54:135–145.CrossRefGoogle Scholar
  2. 2.
    Gunji H, Kharbanda S, Kufe D. Induction of internucleosomal DNA fragmentation in human myeloid leukemia cells by 1-β-D- arabinofuranosylcytosine.Cancer Res. 1991;51:741–743.PubMedGoogle Scholar
  3. 3.
    Shimizu T, Kubota M, Tanizawa A, et al. Inhibition of both etoposide-induced DNA fragmentation and activation of poly(ADP-ribose) synthesis by zinc ion.Biochem Biophy Res Commun. 1990; 169:1172–1177.CrossRefGoogle Scholar
  4. 4.
    Smith CA, Williams GT, Kingston R, Jenkinson EJ, Owen JJT. Antibodies to CD3/T-cell receptor complex induce death by apoptosis in immature T cells in thymic cultures.Nature. 1989;337: 181–184.CrossRefPubMedGoogle Scholar
  5. 5.
    Ucker DS. Cytotoxic T. lymphocytes and glucocorticoids activate an endogenous suicide process in target cells.Nature. 1987;327: 62–64.CrossRefGoogle Scholar
  6. 6.
    MacDonald HR, Lees RK. Programmed death of autoreactive thymocytes.Nature. 1990;343:642–644.CrossRefPubMedGoogle Scholar
  7. 7.
    McConkey DJ, Hartzell P, Amador-Perez JD, Orrenius S, Jondal M. Calcium-dependent killing of immature thymocytes by stimulation via the CD3/T cell receptor complex.J Immunol. 1989;143: 1801–1806.PubMedGoogle Scholar
  8. 8.
    Fruman DA, Mather PE, Burakoff SJ, Bierer BE. Correlation of calcineurin phosphatase activity and programmed cell death in murine T cell hybridomas.Eur J Immunol. 1992;22:2513–2517.CrossRefPubMedGoogle Scholar
  9. 9.
    Okada C, Kizaki H, Tadakuma T. T cell receptor-mediated DNA fragmentation and cell death in T cell hybridomas.J Immunol. 1990;144:2096–2101.Google Scholar
  10. 10.
    Flanagan WM, Corthesy B, Bram RJ, Crabtree GR. Nuclear association of a T-cell transcription factor blocked by FK-506 and cyclosporin A.Nature. 1991;352:803–807.CrossRefPubMedGoogle Scholar
  11. 11.
    Timmerman LA, Clipstone NA, Ho SN, Northrop JP, Crabtree GR. Rapid shuttling of NF-AT in discrimination of Ca2+ signals and immunosuppression.Nature. 1996;383:837–840.CrossRefPubMedGoogle Scholar
  12. 12.
    Reth M. Antigen receptors on B lymphocytes.Annu Rev Immunol. 1992;10:97–121.CrossRefPubMedGoogle Scholar
  13. 13.
    Saouaf SJ, Mahajan S, Rowley RB, et al. Temporal differences in the activation of three classes of non-transmembrane protein tyrosine kinases following B-cell antigen receptor surface engagement.Proc Natl Acad Sci U S A. 1994;91:9524–9528.CrossRefPubMedPubMedCentralGoogle Scholar
  14. 14.
    Rawlings DJ. Bruton’s tyrosine kinase controls a sustained calcium signal essential for B lineage development and function.Clin Immunol. 1999;91:243–253.CrossRefPubMedGoogle Scholar
  15. 15.
    Kurosaki T, Maeda A, Ishiai M, Hashimoto A, Inabe K,Takata M. Regulation of the phospholipase C-γ2 pathway in B cells.Immunol Res. 2000;176:19–29.Google Scholar
  16. 16.
    Dolmetsch RE, Lewis RS, Goodnow CC, Healy JI. Differential activation of transcription factors induced by Ca2+ response amplitude and duration.Nature. 1997;386:855–858.CrossRefPubMedGoogle Scholar
  17. 17.
    Komada Y, Zhang XL, Zhou YW, et al. Apoptotic cell death induced by anti-IgM antibody and phorbol esters is inhibited by interleukin-4 in human B lymphoma cell line MBC-1.Cellular Immunol. 1994;159:280–293.CrossRefGoogle Scholar
  18. 18.
    Pieters R, Leonen AH, Huismans DR, et al.In vitro drug sensitivity of cells from children with leukemia using the MTT assay with improved culture conditions.Blood. 1990;76:2327–2336.PubMedGoogle Scholar
  19. 19.
    Higashigawa M, Komada Y, Shimono Y, et al. FK506 inhibits anti- IgM antibody-induced apoptosis and 17 kD endonuclease activity in the human B-cell line, MBC-1, established from Burkitt’s lymphoma.Br J Haematol. 1997;99:908–913.CrossRefPubMedGoogle Scholar
  20. 20.
    Liu J, Farmer JD Jr, Lane WS, Friedman J, Weissman I, Schreiber SL. Calcineurin is a common target of cyclophilin-cyclosporin A and FKBP-FK506 complexes.Cell. 1991;66:807–815.CrossRefPubMedGoogle Scholar
  21. 21.
    Downward J, Graves J, Cantrell D. The regulation and function of p21ras in T cells.Immunol Today. 1992;13:89–92.CrossRefPubMedGoogle Scholar
  22. 22.
    Healy JI, Dolmetsch RE, Timmerman LA, et al. Different nuclear signals are activated by the B cell receptor during positive versus negative signaling.Immunity. 1997;6:419–428.CrossRefPubMedGoogle Scholar

Copyright information

© The Japanese Society of Hematology 2002

Authors and Affiliations

  1. 1.Department of PediatricsYamada Red Cross HospitalWatarai-gunJapan
  2. 2.Department of PediatricsMie University School of MedicineMieJapan

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