Abstract
The present study demonstrates the minimal, optimal, and synergistic signals involved in the activation of normal human peripheral blood and tonsillar B cells to proliferation. Initial activation signals were delivered to B cells by low concentrations of anti-mu antibody which did not induce proliferation by themselves. However, marked synergy was seen when anti-mu antibody was added to cultures in the presence of monoclonal B cell growth factor (BCGF) obtained from a human T-T cell hybrid such that the B cells underwent substantial proliferation. This latter proliferation was seen without maturation into Ig-secreting cells, which indicates that the BCGF is not a differentiation signal but a signal that drives the cell up to but not beyond the proliferative phase. Of note was the fact that B cells reflected differential sensitivity on the basis of size to either the activation signal delivered by anti-mu antibody or the proliferative signal delivered by BCGF. BCGF directly stimulated the larger B cells in the normal tonsillar B cell repertoire to proliferate without the requirement for an in vitro activation signal, which indicates that the cells had already received some form of activation signal in vivo. Indeed, these cells expressed the 4F2 antigen found on activated but not resting lymphocytes. In contrast, the smaller tonsillar B lymphocytes did not express the 4F2 activation antigen and required activation by anti-mu antibody, which did not of itself induce proliferation, but which acted in synergy with BCGF for substantial proliferation of the B cells. These studies thus provide a useful model of human B cell activation, proliferation, and differentiation and allow a more precise delineation of each phase in this cascade.
Full Text
The Full Text of this article is available as a PDF (1.1 MB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Adinolfi M., Gardner B., Giannelli F., McGuire M. Studies on human lymphocytes stimulated in vitro with anti-gamma and anti-mu antibodies. Experientia. 1967 Apr 15;23(4):271–272. doi: 10.1007/BF02135679. [DOI] [PubMed] [Google Scholar]
- 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]
- Baker P. E., Gillis S., Smith K. A. Monoclonal cytolytic T-cell lines. J Exp Med. 1979 Jan 1;149(1):273–278. doi: 10.1084/jem.149.1.273. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bloch-Shtacher N., Hirschhorn K., Uhr J. W. The response of lymphocytes from non-immunized humans to antigen-antibody complexes. Clin Exp Immunol. 1968 Nov;3(9):889–899. [PMC free article] [PubMed] [Google Scholar]
- Butler J. L., Muraguchi A., Lane H. C., Fauci A. S. Development of a human T-T cell hybridoma secreting B cell growth factor. J Exp Med. 1983 Jan 1;157(1):60–68. doi: 10.1084/jem.157.1.60. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Chiorazzi N., Fu S. M., Kunkel H. G. Stimulation of human B lymphocytes by antibodies to IgM and IgG: functional evidence for the expression of IgG on B-lymphocyte surface membranes. Clin Immunol Immunopathol. 1980 Mar;15(3):301–313. doi: 10.1016/0090-1229(80)90042-2. [DOI] [PubMed] [Google Scholar]
- Contreras T. J., Jemionek J. F., Stevenson H. C., Hartwig V. M., Fauci A. S. An improved technique for the negative selection of large numbers of human lymphocytes and monocytes by counterflow centrifugation--elutriation. Cell Immunol. 1980 Aug 15;54(1):215–229. doi: 10.1016/0008-8749(80)90203-8. [DOI] [PubMed] [Google Scholar]
- 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]
- 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]
- Dutton R. W. Separate signals for the initiation of proliferation and differentiation in the b cell response to antigen. Transplant Rev. 1975;23:66–77. doi: 10.1111/j.1600-065x.1975.tb00149.x. [DOI] [PubMed] [Google Scholar]
- Eisenbarth G. S., Haynes B. F., Schroer J. A., Fauci A. S. Production of monoclonal antibodies reacting with peripheral blood mononuclear cell surface differentiation antigens. J Immunol. 1980 Mar;124(3):1237–1244. [PubMed] [Google Scholar]
- Falkoff R. J., Zhu L. P., Fauci A. S. Separate signals for human B cell proliferation and differentiation in response to Staphylococcus aureus: evidence for a two-signal model of B cell activation. J Immunol. 1982 Jul;129(1):97–102. [PubMed] [Google Scholar]
- Farrar J. J., Benjamin W. R., Hilfiker M. L., Howard M., Farrar W. L., Fuller-Farrar J. The biochemistry, biology, and role of interleukin 2 in the induction of cytotoxic T cell and antibody-forming B cell responses. Immunol Rev. 1982;63:129–166. doi: 10.1111/j.1600-065x.1982.tb00414.x. [DOI] [PubMed] [Google Scholar]
- Farrar J. J., Fuller-Farrar J., Simon P. L., Hilfiker M. L., Stadler B. M., Farrar W. L. Thymoma production of T cell growth factor (Interleukin 2). J Immunol. 1980 Dec;125(6):2555–2558. [PubMed] [Google Scholar]
- Fauci A. S., Whalen G., Burch C. Activation of human B lymphocytes XVI. Cellular requirements, interactions, and immunoregulation of pokeweed mitogen-induced total-immunoglobulin producing plaque-forming cells in peripheral blood. Cell Immunol. 1980 Aug 15;54(1):230–240. doi: 10.1016/0008-8749(80)90204-x. [DOI] [PubMed] [Google Scholar]
- Gausset P., Delespesse G., Hubert C., Kennes B., Govaerts A. In vitro response of subpopulations of human lymphocytes. II. DNA synthesis induced by anti-immunoglobulin antibodies. J Immunol. 1976 Feb;116(2):446–453. [PubMed] [Google Scholar]
- Gerrard T. L., Fauci A. S. Activation and immunoregulation of antigen-specific human b lymphocyte responses: multifaceted role of the monocyte. J Immunol. 1982 May;128(5):2367–2372. [PubMed] [Google Scholar]
- Gillis S., Mizel S. B. T-Cell lymphoma model for the analysis of interleukin 1-mediated T-cell activation. Proc Natl Acad Sci U S A. 1981 Feb;78(2):1133–1137. doi: 10.1073/pnas.78.2.1133. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gillis S., Smith K. A. Long term culture of tumour-specific cytotoxic T cells. Nature. 1977 Jul 14;268(5616):154–156. doi: 10.1038/268154a0. [DOI] [PubMed] [Google Scholar]
- Haynes B. F., Hemler M. E., Mann D. L., Eisenbarth G. S., Shelhamer J., Mostowski H. S., Thomas C. A., Strominger J. L., Fauci A. S. Characterization of a monoclonal antibody (4F2) that binds to human monocytes and to a subset of activated lymphocytes. J Immunol. 1981 Apr;126(4):1409–1414. [PubMed] [Google Scholar]
- Howard M., Farrar J., Hilfiker M., Johnson B., Takatsu K., Hamaoka T., Paul W. E. Identification of a T cell-derived b cell growth factor distinct from interleukin 2. J Exp Med. 1982 Mar 1;155(3):914–923. doi: 10.1084/jem.155.3.914. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Howard M., Kessler S., Chused T., Paul W. E. Long-term culture of normal mouse B lymphocytes. Proc Natl Acad Sci U S A. 1981 Sep;78(9):5788–5792. doi: 10.1073/pnas.78.9.5788. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kishimoto T., Ishizaka K. Regulation of antibody response in vitro. IX. Induction of secondary anti-hapten IgG antibody response by anti-immunoglobulin and enhancing soluble factor. J Immunol. 1975 Feb;114(2 Pt 1):585–591. [PubMed] [Google Scholar]
- Kishimoto T., Miyake T., Nishizawa Y., Watanabe T., Yamamura Y. Triggering mechanism of B lymphocytes. I. Effect of anti-immunoglobulin and enhancing soluble factor on differentiation and proliferation of B cells. J Immunol. 1975 Nov;115(5):1179–1184. [PubMed] [Google Scholar]
- Lane H. C., Volkman D. J., Whalen G., Fauci A. S. In vitro antigen-induced, antigen-specific antibody production in man. Specific and polyclonal components, kinetics, and cellular requirements. J Exp Med. 1981 Oct 1;154(4):1043–1057. doi: 10.1084/jem.154.4.1043. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Muraguchi A., Fauci A. S. Proliferative responses of normal human B lymphocytes. Development of an assay system for human B cell growth factor (BCGF). J Immunol. 1982 Sep;129(3):1104–1108. [PubMed] [Google Scholar]
- Muraguchi A., Kasahara T., Oppenheim J. J., Fauci A. S. B cell growth factor and T cell growth factor produced by mitogen-stimulated normal human peripheral blood T lymphocytes are distinct molecules. J Immunol. 1982 Dec;129(6):2486–2489. [PubMed] [Google Scholar]
- Muraguchi A., Kishimoto T., Kuritani T., Watanabe T., Yamamura Y. In vitro immune response of human peripheral lymphocytes. V. PHA- and protein A-induced human B colony formation and analysis of the subpopulations of B cells. J Immunol. 1980 Aug;125(2):564–569. [PubMed] [Google Scholar]
- Muraguchi A., Kishimoto T., Miki Y., Kuritani T., Kaieda T., Yoshizaki K., Yamamura Y. T cell-replacing factor- (TRF) induced IgG secretion in a human B blastoid cell line and demonstration of acceptors for TRF. J Immunol. 1981 Aug;127(2):412–416. [PubMed] [Google Scholar]
- Oppenheim J. J., Rogentine G. N., Terry W. D. The transformation of human lymphocytes by monkey antisera to human immunoglobulins. Immunology. 1969 Jan;16(1):123–138. [PMC free article] [PubMed] [Google Scholar]
- Oppenheim J. J., Stadler B. M., Siraganian R. P., Mage M., Mathieson B. Lymphokines: their role in lymphocyte responses. Properties of interleukin 1. Fed Proc. 1982 Feb;41(2):257–262. [PubMed] [Google Scholar]
- Parker D. C., Fothergill J. J., Wadsworth D. C. B lymphocyte activation by insoluble anti-immunoglobulin: induction of immunoglobulin secretion by a T cell-dependent soluble factor. J Immunol. 1979 Aug;123(2):931–941. [PubMed] [Google Scholar]
- Romagnani S., Giudizi M. G., Almerigogna F., Ricci M. Interaction of staphylococcal protein A with membrane components of IgM- and/or IgD-bearing lymphocytes from human tonsil. J Immunol. 1980 Apr;124(4):1620–1626. [PubMed] [Google Scholar]
- SELL S., GELL P. G. STUDIES ON RABBIT LYMPHOCYTES IN VITRO. I. STIMULATION OF BLAST TRANSFORMATION WITH AN ANTIALLOTYPE SERUM. J Exp Med. 1965 Aug 1;122:423–440. doi: 10.1084/jem.122.2.423. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Schrier R. D., Skidmore B. J., Kurnick J. T., Goldstine S. N., Chiller J. M. Propagation of antigen-specific T cell helper function in vitro. J Immunol. 1979 Dec;123(6):2525–2531. [PubMed] [Google Scholar]
- Sieckmann D. G., Asofsky R., Mosier D. E., Zitron I. M., Paul W. E. Activation of mouse lymphocytes by anti-immunoglobulin. I. Parameters of the proliferative response. J Exp Med. 1978 Mar 1;147(3):814–829. doi: 10.1084/jem.147.3.814. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sieckmann D. G., Scher I., Asofsky R., Mosier D. E., Paul W. E. Activation of mouse lymphocytes by anti-immunoglobulin. II. A thymus-independent response by a mature subset of B lymphocytes. J Exp Med. 1978 Dec 1;148(6):1628–1643. doi: 10.1084/jem.148.6.1628. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sredni B., Sieckmann D. G., Kumagai S., House S., Green I., Paul W. E. Long-term culture and cloning of nontransformed human B lymphocytes. J Exp Med. 1981 Nov 1;154(5):1500–1516. doi: 10.1084/jem.154.5.1500. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sredni B., Tse H. Y., Schwartz R. H. Direct cloning and extended culture of antigen-specific MHC-restricted, proliferating T lymphocytes. Nature. 1980 Feb 7;283(5747):581–583. doi: 10.1038/283581a0. [DOI] [PubMed] [Google Scholar]
- Sredni B., Volkman D., Schwartz R. H., Fauci A. S. Antigen-specific human T-cell clones: development of clones requiring HLA-DR-compatible presenting cells for stimulation in presence of antigen. Proc Natl Acad Sci U S A. 1981 Mar;78(3):1858–1862. doi: 10.1073/pnas.78.3.1858. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Teranishi T., Hirano T., Arima N., Onoue K. Human helper T cell factor(s) (ThF). II. Induction of IgG production in B lymphoblastoid cell lines and identification of T cell-replacing factor- (TRF) like factor(s). J Immunol. 1982 Apr;128(4):1903–1908. [PubMed] [Google Scholar]
- Watanabe T., Yoshizaki K., Yagura T., Yamamura Y. In vitro antibody formation by human tonsil lymphocytes. J Immunol. 1974 Aug;113(2):608–616. [PubMed] [Google Scholar]
- Watson J. Continuous proliferation of murine antigen-specific helper T lymphocytes in culture. J Exp Med. 1979 Dec 1;150(6):1510–1519. doi: 10.1084/jem.150.6.1510. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Whitlock C. A., Witte O. N. Long-term culture of B lymphocytes and their precursors from murine bone marrow. Proc Natl Acad Sci U S A. 1982 Jun;79(11):3608–3612. doi: 10.1073/pnas.79.11.3608. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Yoshizaki K., Nakagawa T., Kaieda T., Muraguchi A., Yamamura Y., Kishimoto T. Induction of proliferation and Ig production in human B leukemic cells by anti-immunoglobulins and T cell factors. J Immunol. 1982 Mar;128(3):1296–1301. [PubMed] [Google Scholar]