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Clinical and Experimental Immunology logoLink to Clinical and Experimental Immunology
. 1985 Sep;61(3):648–656.

Analysis of the antigen-induced in vitro differentiation of human peripheral blood B cells: stimulation with ovalbumin induces the transition from the resting state into the excited state.

L A Hart, C J Heijnen, J Zijlstra, R E Ballieux
PMCID: PMC1577281  PMID: 2934190

Abstract

Human peripheral blood B lymphocytes, when cultured for 6 days with ovalbumin in the presence of T cells and monocytes, can differentiate into plaque-forming cells (PFC) secreting small amounts of antigen-specific IgM. Phenotype analysis using Ig-isotype and B-cell specific monoclonal antibodies revealed that the PFC precursor is a sIgM+ IgD+IgG- B lymphocyte. The PFC itself also carries the B-cell specific membrane antigens B1 and BA-1; it expresses only IgM on its surface. When apart from the antigen a Concanavalin A-induced, T-cell derived growth factor preparation was added to the cultures, we observed the disappearance of sIgM from the membrane. Furthermore, it was demonstrated by means of density gradient centrifugation that the differentiation of the PFC-precursor into a PFC is accompanied by a decrease in cellular density. Finally, it was shown using hydroxyurea as an inhibitor of DNA-synthesis, that the PFC-precursor passes several cycles of cell division during the process of differentiation into a PFC.

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Selected References

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  1. Abramson C. S., Kersey J. H., LeBien T. W. A monoclonal antibody (BA-1) reactive with cells of human B lymphocyte lineage. J Immunol. 1981 Jan;126(1):83–88. [PubMed] [Google Scholar]
  2. Anderson J., Melchers F. T cell-dependent activation of resting B cells: requirement for both nonspecific unrestricted and antigen-specific Ia-restricted soluble factors. Proc Natl Acad Sci U S A. 1981 Apr;78(4):2497–2501. doi: 10.1073/pnas.78.4.2497. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Andersson J., Coutinho A., Lernhardt W., Melchers F. Clonal growth and maturation to immunoglobulin secretion in vitro of every growth-inducible B lymphocyte. Cell. 1977 Jan;10(1):27–34. doi: 10.1016/0092-8674(77)90136-2. [DOI] [PubMed] [Google Scholar]
  4. Ballieux R. E., Heijnen C. J. Immunoregulatory T cell subpopulations in man: dissection by monoclonal antibodies and Fc-receptors. Immunol Rev. 1983;74:5–28. doi: 10.1111/j.1600-065x.1983.tb01082.x. [DOI] [PubMed] [Google Scholar]
  5. Ballieux R. E., Heijnen C. J., Uytdehaag F., Zegers B. J. Regulation of B cell activity in man: role of T cells. Immunol Rev. 1979;45:3–39. doi: 10.1111/j.1600-065x.1979.tb00271.x. [DOI] [PubMed] [Google Scholar]
  6. DeFranco A. L., Kung J. T., Paul W. E. Regulation of growth and proliferation in B cell subpopulations. Immunol Rev. 1982;64:161–182. doi: 10.1111/j.1600-065x.1982.tb00423.x. [DOI] [PubMed] [Google Scholar]
  7. Dosch H-M, Gelfand E. W. Generation of human plaque-forming cells in culture: tissue distribution, antigenic and cellular requirements. J Immunol. 1977 Jan;118(1):302–308. [PubMed] [Google Scholar]
  8. Dutton R. W., Mishell R. I. Cell populations and cell proliferation in the in vitro response of normal mouse spleen to heterologous erythrocytes. Analysis by the hot pulse technique. J Exp Med. 1967 Sep 1;126(3):443–454. doi: 10.1084/jem.126.3.443. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Gobbi M., Caligaris-Cappio F., Janossy G. Normal equivalent cells of B cell malignancies: analysis with monoclonal antibodies. Br J Haematol. 1983 Jul;54(3):393–403. doi: 10.1111/j.1365-2141.1983.tb02114.x. [DOI] [PubMed] [Google Scholar]
  10. Goding J. W. The chromic chloride method of coupling antigens to erythrocytes: definition of some important parameters. J Immunol Methods. 1976;10(1):61–66. doi: 10.1016/0022-1759(76)90007-7. [DOI] [PubMed] [Google Scholar]
  11. Heijnen C. J., Pot K. H., Ballieux R. E. Characterization of human T suppressor-inducer, -precursor and -effector lymphocytes in the antigen-specific plaque-forming cell response. Eur J Immunol. 1982 Oct;12(10):860–866. doi: 10.1002/eji.1830121012. [DOI] [PubMed] [Google Scholar]
  12. Heijnen C. J., UytdeHaag F., Ballieux R. E. In vitro antibody response of human lymphocytes. Springer Semin Immunopathol. 1980 May;3(1):63–92. doi: 10.1007/BF00199926. [DOI] [PubMed] [Google Scholar]
  13. Heijnen C. J., UytdeHaag F., Gmelig-Meyling F. H., Ballieux R. E. Localization of human antigen-specific helper and suppressor function in distinct T-cell subpopulations. Cell Immunol. 1979 Mar 15;43(2):282–292. doi: 10.1016/0008-8749(79)90173-4. [DOI] [PubMed] [Google Scholar]
  14. Howard M., Paul W. E. Regulation of B-cell growth and differentiation by soluble factors. Annu Rev Immunol. 1983;1:307–333. doi: 10.1146/annurev.iy.01.040183.001515. [DOI] [PubMed] [Google Scholar]
  15. Jerne N. K., Henry C., Nordin A. A., Fuji H., Koros A. M., Lefkovits I. Plaque forming cells: methodology and theory. Transplant Rev. 1974;18:130–191. doi: 10.1111/j.1600-065x.1974.tb01588.x. [DOI] [PubMed] [Google Scholar]
  16. Melchers F., Andersson J., Corbel C., Leptin M., Lernhardt W., Gerhard W., Zeuthen J. Regulation of B lymphocyte replication and maturation. J Cell Biochem. 1982;19(4):315–332. doi: 10.1002/jcb.240190403. [DOI] [PubMed] [Google Scholar]
  17. Michlmayr G., Huber H. Receptor sites for complement on certain human peripheral blood lymphocytes. J Immunol. 1970 Sep;105(3):670–676. [PubMed] [Google Scholar]
  18. Misiti J., Waldmann T. A. In vitro generation of antigen-specific hemolytic plaque-forming cells from human peripheral blood mononuclear cells. J Exp Med. 1981 Oct 1;154(4):1069–1084. doi: 10.1084/jem.154.4.1069. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Monroe J. G., Havran W. L., Cambier J. C. B lymphocyte activation: entry into cell cycle is accompanied by decreased expression of IgD but not IgM. Eur J Immunol. 1983 Mar;13(3):208–213. doi: 10.1002/eji.1830130306. [DOI] [PubMed] [Google Scholar]
  20. Muchmore A. V., Koski I., Dooley N., Blaese R. M. Artifactual plaque formation in vitro and in vivo to passive transfer of specific antibody. J Immunol. 1976 Apr;116(4):1016–1019. [PubMed] [Google Scholar]
  21. Stashenko P., Nadler L. M., Hardy R., Schlossman S. F. Characterization of a human B lymphocyte-specific antigen. J Immunol. 1980 Oct;125(4):1678–1685. [PubMed] [Google Scholar]
  22. Wrigley D. M., Choi Y. S. Characterization of the subpopulations of human peripheral blood B lymphocytes which react to staphylococcus aureus Cowan I and T-cell help. Cell Immunol. 1983 May;78(1):130–143. doi: 10.1016/0008-8749(83)90266-6. [DOI] [PubMed] [Google Scholar]
  23. de Gast G. C., Platts-Mills T. A. Functional studies on lymphocytes in adult human bone marrow. II. Isolated surface IgM-positive cells. J Immunol. 1979 Jan;122(1):285–290. [PubMed] [Google Scholar]

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