Skip to main content
PMC home page
Immunology logoLink to Immunology
. 1979 Sep;38(1):181–189.

Pokeweed mitogen induced differentiation of human B cells: evaluation by a protein A haemolytic plaque assay.

L Hammarström, A G Bird, S Britton, C I Smith
PMCID: PMC1457881  PMID: 116958

Abstract

Using the protein A plaque assay, the number of human cells secreting immunoglobulin of various classes after pokeweed mitogen stimulation was determined. At optimal response (on day 5-7) a mean of 58,354 IgM PFC/10(6), 34,207 IgG PFC/10(6) and 10,525 IgA PFC/10(6) cells was found when using peripheral blood lymphocytes. In spleen cells, peak values which were slightly higher than in blood, were obtained at day 4-6. The proportions of cells secreting light chains of either type were found to be comparable to those of unstimulated cells thus supporting the notion of the polyclonality of the response. Pokeweed mitogen stimulation of peripheral blood lymphocytes was found to be totally T-cell dependent whereas the response of spleen cells was not. When assayed for antigen-specific precursor cells in cultures stimulated by mitogen, the frequency of SRBC-specific IgM producing cells was found to be 1.3/1000 cells. This frequency was regularly found to be independent of medium supplement.

Full text

PDF
181

Selected References

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

  1. Andersson J., Coutinho A., Melchers F. Frequencies of mitogen-reactive B cells in the mouse. II. Frequencies of B cells producing antibodies which lyse sheep or horse erythrocytes, and trinitrophenylated or nitroiodophenylated sheep erythrocytes. J Exp Med. 1977 Jun 1;145(6):1520–1530. doi: 10.1084/jem.145.6.1520. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bird A. G., Britton S. A live human B-cell activator operating in isolation of other cellular influences. Scand J Immunol. 1979;9(6):507–510. doi: 10.1111/j.1365-3083.1979.tb03278.x. [DOI] [PubMed] [Google Scholar]
  3. Dosch H. M., Percy M. E., Gelfand E. W. Functional differentiation of B lymphocytes in congenital agammaglobulinemia. I. Generation of hemolytic plaque-forming cells. J Immunol. 1977 Dec;119(6):1959–1964. [PubMed] [Google Scholar]
  4. Fauci A. S., Pratt K. R. Activation of human B lymphocytes. I. Direct plaque-forming cell assay for the measurement of polyclonal activation and antigenic stimulation of human B lymphocytes. J Exp Med. 1976 Sep 1;144(3):674–684. doi: 10.1084/jem.144.3.674. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Fauci A. S., Pratt K. R. Polyclonal activation of bone-marrow-derived lymphocytes from human peripheral blood measured by a direct plaque-forming cell assay. Proc Natl Acad Sci U S A. 1976 Oct;73(10):3676–3679. doi: 10.1073/pnas.73.10.3676. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Finkelman F. D., Lipsky P. E. Immunoglobulin secretion by human splenic lymphocytes in vitro: the effects of antibodies to IgM and IgD. J Immunol. 1978 May;120(5):1465–1472. [PubMed] [Google Scholar]
  7. Ginsburg W. W., Finkelman F. D., Lipsky P. E. Circulating and mitogen-induced immunoglobulin-secreting cells in human peripheral blood: evaluation by a modified reverse hemolytic plaque assay. J Immunol. 1978 Jan;120(1):33–39. [PubMed] [Google Scholar]
  8. Greaves M., Janossy G., Doenhoff M. Selective triggering of human T and B lymphocytes in vitro by polyclonal mitogens. J Exp Med. 1974 Jul 1;140(1):1–18. doi: 10.1084/jem.140.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Gronowicz E., Coutinho A., Melchers F. A plaque assay for all cells secreting Ig of a given type or class. Eur J Immunol. 1976 Aug;6(8):588–590. doi: 10.1002/eji.1830060812. [DOI] [PubMed] [Google Scholar]
  10. Ivanyi L., Lehner T. Stimulation of human lymphocytes by B-cell mitogens. Clin Exp Immunol. 1974 Nov;18(3):347–356. [PMC free article] [PubMed] [Google Scholar]
  11. Janossy G., Gomez de la Concha E., Luquetti A., Snajdr M. J., Waxdal M. J., Platts-Mills T. A. T-cell regulation of immunoglobulin synthesis and proliferation in pokeweed (Pa-1)-stimulated human lymphocyte cultures. Scand J Immunol. 1977;6(1-2):109–123. doi: 10.1111/j.1365-3083.1977.tb00326.x. [DOI] [PubMed] [Google Scholar]
  12. Janossy G., Greaves M. Functional analysis of murine and human B lymphocyte subsets. Transplant Rev. 1975;24:177–236. doi: 10.1111/j.1600-065x.1975.tb00169.x. [DOI] [PubMed] [Google Scholar]
  13. Jerne N. K., Nordin A. A. Plaque Formation in Agar by Single Antibody-Producing Cells. Science. 1963 Apr 26;140(3565):405–405. doi: 10.1126/science.140.3565.405. [DOI] [PubMed] [Google Scholar]
  14. Julius M. H., Simpson E., Herzenberg L. A. A rapid method for the isolation of functional thymus-derived murine lymphocytes. Eur J Immunol. 1973 Oct;3(10):645–649. doi: 10.1002/eji.1830031011. [DOI] [PubMed] [Google Scholar]
  15. Keightley R. G., Cooper M. D., Lawton A. R. The T cell dependence of B cell differentiation induced by pokeweed mitogen. J Immunol. 1976 Nov;117(5 Pt 1):1538–1544. [PubMed] [Google Scholar]
  16. OPPENHEIM J. J., PERRY S. EFFECTS OF ENDOTOXINS ON CULTURED LEUKOCYTES. Proc Soc Exp Biol Med. 1965 Apr;118:1014–1019. doi: 10.3181/00379727-118-30033. [DOI] [PubMed] [Google Scholar]
  17. Persson U., Bick P. H., Hammarströme L., Möller E., Smith C. I. Different requirements for T cells responding to various doses of concanavalin A. Scand J Immunol. 1978;8(4):291–301. doi: 10.1111/j.1365-3083.1978.tb00522.x. [DOI] [PubMed] [Google Scholar]
  18. Ringdén O., Rynnel-Dagö B., Kunori T., Freijd A., Möller E. Antibody production and DNA synthesis of human lymphocyte subpopulations induced by PPD tuberculin. Clin Exp Immunol. 1979 Jun;36(3):528–535. [PMC free article] [PubMed] [Google Scholar]
  19. Ringdén O., Rynnel-Dagö B., Waterfield E. M., Möller E., Möller G. Polyclonal antibody secretion in human lymphocytes induced by killed staphylococcal bacteria and by lipopolysaccharide. Scand J Immunol. 1977;6(11):1159–1169. doi: 10.1111/j.1365-3083.1977.tb00355.x. [DOI] [PubMed] [Google Scholar]
  20. Smith C. I., Hammarström L. Regulation of lymphocyte activation by serum factors. Int Arch Allergy Appl Immunol. 1979;58(2):219–226. doi: 10.1159/000232195. [DOI] [PubMed] [Google Scholar]
  21. Waxdal M. J., Basham T. Y. B and T-cell stimulatory activities of multiple mitogens from pokeweed. Nature. 1974 Sep 13;251(5471):163–164. doi: 10.1038/251163a0. [DOI] [PubMed] [Google Scholar]

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

RESOURCES