Skip to main content
PMC home page
Immunology logoLink to Immunology
. 1989 Jul;67(3):359–364.

Differential responsiveness of human B lymphocytes to phorbol ester and calcium ionophore based on their state of activation.

J Inglés 1, P Engel 1, O De La Calle 1, T Gallart 1
PMCID: PMC1385353  PMID: 2503437

Abstract

For tonsil B cells of a particular high density (below 65% Percoll), both phorbol myristate acetate (PMA) (5 ng/ml) and calcium ionophore A23187 (500 nM) were required to induce RNA synthesis, significant DNA synthesis also occurring in the presence of 12,000 MW B-cell growth factor (BCGF). In contrast, PMA alone, even at 1 ng/ml, was a sufficient stimulus to induce strong DNA synthesis in low-density B cells (45-50% Percoll) and strong proliferative responsiveness to BCGF in intermediate-density B cells (55-65% Percoll). In these latter B-cell populations, A23187 (500 nM), acted synergistically with non-mitogenic PMA doses to induce strong DNA synthesis, the PMA dose required being 5-50 times lower in low-density B cells (0.1-1 ng/ml) than in intermediate-density B cells (5 ng/ml). Preactivation for 30 hr with anti-Ig antibodies plus BCGF, known to drive B cells into late G1, rendered high-density B cells responsive to PMA (1-10 ng/ml) with high, dose-related DNA synthesis. These data indicate that the B-cell mitogenicity of a given nanomolar dose of PMA depends on the more advanced state of activation of B cells. It was also found that the above optimal dose of A23187 (500 nM) paradoxically inhibited the PMA-induced DNA synthesis of low-density B cells and in vitro preactivated high-density B cells. Data obtained with low-density B cells suggest that a calcium influx during the PMA-induced proliferative phase of B cells may provide a negative signal for the DNA synthesis.

Full text

PDF
359

Selected References

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

  1. Bijsterbosch M. K., Meade C. J., Turner G. A., Klaus G. G. B lymphocyte receptors and polyphosphoinositide degradation. Cell. 1985 Jul;41(3):999–1006. doi: 10.1016/s0092-8674(85)80080-5. [DOI] [PubMed] [Google Scholar]
  2. Boyd A. W., Metcalf D. Activation of human and murine B cells by anti-immunoglobulin antibody: dependence of the response on the preexisting level of B-cell activation. Cell Immunol. 1984 Nov;89(1):75–83. doi: 10.1016/0008-8749(84)90199-0. [DOI] [PubMed] [Google Scholar]
  3. Chen Z. Z., Coggeshall K. M., Cambier J. C. Translocation of protein kinase C during membrane immunoglobulin-mediated transmembrane signaling in B lymphocytes. J Immunol. 1986 Mar 15;136(6):2300–2304. [PubMed] [Google Scholar]
  4. Clark E. A., Shu G. Activation of human B cell proliferation through surface Bp35 (CD20) polypeptides or immunoglobulin receptors. J Immunol. 1987 Feb 1;138(3):720–725. [PubMed] [Google Scholar]
  5. Defranco A. L., Raveche E. S., Asofsky R., Paul W. E. Frequency of B lymphocytes responsive to anti-immunoglobulin. J Exp Med. 1982 May 1;155(5):1523–1536. doi: 10.1084/jem.155.5.1523. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Guy G. R., Bunce C. M., Gordon J., Michell R. H., Brown G. A combination of calcium ionophore and 12-O-tetradecanoyl-phorbol-13-acetate (TPA) stimulates the growth of purified resting B cells. Scand J Immunol. 1985 Nov;22(5):591–596. doi: 10.1111/j.1365-3083.1985.tb01919.x. [DOI] [PubMed] [Google Scholar]
  7. Hayakawa K., Hardy R. R., Stall A. M., Herzenberg L. A., Herzenberg L. A. Immunoglobulin-bearing B cells reconstitute and maintain the murine Ly-1 B cell lineage. Eur J Immunol. 1986 Oct;16(10):1313–1316. doi: 10.1002/eji.1830161021. [DOI] [PubMed] [Google Scholar]
  8. Kehrl J. H., Muraguchi A., Fauci A. S. Human B cell activation and cell cycle progression: stimulation with anti-mu and Staphylococcus aureus Cowan strain I. Eur J Immunol. 1984 Feb;14(2):115–121. doi: 10.1002/eji.1830140203. [DOI] [PubMed] [Google Scholar]
  9. Klaus G. G., O'Garra A., Bijsterbosch M. K., Holman M. Activation and proliferation signals in mouse B cells. VIII. Induction of DNA synthesis in B cells by a combination of calcium ionophores and phorbol myristate acetate. Eur J Immunol. 1986 Jan;16(1):92–97. doi: 10.1002/eji.1830160118. [DOI] [PubMed] [Google Scholar]
  10. Ledbetter J. A., Rabinovitch P. S., June C. H., Song C. W., Clark E. A., Uckun F. M. Antigen-independent regulation of cytoplasmic calcium in B cells with a 12-kDa B-cell growth factor and anti-CD19. Proc Natl Acad Sci U S A. 1988 Mar;85(6):1897–1901. doi: 10.1073/pnas.85.6.1897. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Maizel A., Sahasrabuddhe C., Mehta S., Morgan J., Lachman L., Ford R. Biochemical separation of a human B cell mitogenic factor. Proc Natl Acad Sci U S A. 1982 Oct;79(19):5998–6002. doi: 10.1073/pnas.79.19.5998. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. May W. S., Jr, Sahyoun N., Wolf M., Cuatrecasas P. Role of intracellular calcium mobilization in the regulation of protein kinase C-mediated membrane processes. Nature. 1985 Oct 10;317(6037):549–551. doi: 10.1038/317549a0. [DOI] [PubMed] [Google Scholar]
  13. Mehta S. R., Conrad D., Sandler R., Morgan J., Montagna R., Maizel A. L. Purification of human B cell growth factor. J Immunol. 1985 Nov;135(5):3298–3302. [PubMed] [Google Scholar]
  14. Mehta S. R., Grant S. R., Maizel A. L. Characterization of the cell surface receptors for human B cell growth factor of 12,000 molecular weight. J Immunol. 1986 Oct 1;137(7):2210–2214. [PubMed] [Google Scholar]
  15. Mold C., Cooper N. R., Nemerow G. R. Incorporation of the purified Epstein Barr virus/C3d receptor (CR2) into liposomes and demonstration of its dual ligand binding functions. J Immunol. 1986 Jun 1;136(11):4140–4145. [PubMed] [Google Scholar]
  16. Monroe J. G., Cambier J. C. Cell cycle dependence for expression of membrane associated IgD, IgM and Ia antigen on mitogen-stimulated murine B-lymphocytes. Ann N Y Acad Sci. 1982;399:238–254. doi: 10.1111/j.1749-6632.1982.tb25677.x. [DOI] [PubMed] [Google Scholar]
  17. Monroe J. G., Kass M. J. Molecular events in B cell activation. I. Signals required to stimulate G0 to G1 transition of resting B lymphocytes. J Immunol. 1985 Sep;135(3):1674–1682. [PubMed] [Google Scholar]
  18. Muraguchi A., Butler J. L., Kehrl J. H., Fauci A. S. Differential sensitivity of human B cell subsets to activation signals delivered by anti-mu antibody and proliferative signals delivered by a monoclonal B cell growth factor. J Exp Med. 1983 Feb 1;157(2):530–546. doi: 10.1084/jem.157.2.530. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Roifman C. M., Benedict S. H., Cheung R. K., Gelfand E. W. Induction of human B cell proliferation and differentiation by the combination of phorbol ester and ionomycin. Eur J Immunol. 1987 May;17(5):701–706. doi: 10.1002/eji.1830170519. [DOI] [PubMed] [Google Scholar]
  20. Sharma S., Mehta S., Morgan J., Maizel A. Molecular cloning and expression of a human B-cell growth factor gene in Escherichia coli. Science. 1987 Mar 20;235(4795):1489–1492. doi: 10.1126/science.3547651. [DOI] [PubMed] [Google Scholar]
  21. Thompson C. B., Scher I., Schaefer M. E., Lindsten T., Finkelman F. D., Mond J. J. Size-dependent B lymphocyte subpopulations: relationship of cell volume to surface phenotype, cell cycle, proliferative response, and requirements for antibody production to TNP-Ficoll and TNP-BA. J Immunol. 1984 Nov;133(5):2333–2342. [PubMed] [Google Scholar]
  22. Walker L., Guy G., Brown G., Rowe M., Milner A. E., Gordon J. Control of human B-lymphocyte replication. I. Characterization of novel activation states that precede the entry of G0 B cells into cycle. Immunology. 1986 Aug;58(4):583–589. [PMC free article] [PubMed] [Google Scholar]
  23. Wilson H. A., Greenblatt D., Taylor C. W., Putney J. W., Tsien R. Y., Finkelman F. D., Chused T. M. The B lymphocyte calcium response to anti-Ig is diminished by membrane immunoglobulin cross-linkage to the Fc gamma receptor. J Immunol. 1987 Mar 15;138(6):1712–1718. [PubMed] [Google Scholar]
  24. Wolf M., LeVine H., 3rd, May W. S., Jr, Cuatrecasas P., Sahyoun N. A model for intracellular translocation of protein kinase C involving synergism between Ca2+ and phorbol esters. Nature. 1985 Oct 10;317(6037):546–549. doi: 10.1038/317546a0. [DOI] [PubMed] [Google Scholar]
  25. Wyllie A. H. Glucocorticoid-induced thymocyte apoptosis is associated with endogenous endonuclease activation. Nature. 1980 Apr 10;284(5756):555–556. doi: 10.1038/284555a0. [DOI] [PubMed] [Google Scholar]

Articles from Immunology are provided here courtesy of British Society for Immunology

RESOURCES