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Molecular and Quantitative Analysis of Helper T Cell-Replacing Factors on the Induction of Antigen-Sensitive B and T Lymphocytes

From the Department of Microbiology, University of California, Irvine College of Medicine, Irvine, California 92717, the Basel Institute for Immunology, Grenzacherstrasse 487, Postfach, 4005 Basel 5, Switzerland, and the Department of Biochemistry, University of Alberta, Edmonton, Alberta, T6G 2H7 Canada

Abstract

Murine spleen cells activated by the T cell mitogen concanavalin A (Con A) secrete factors that exhibit biologic activity on both bone-marrow (B) and thymus-derived (T) lymphocytes. In the presence of heterologous erythrocyte antigens, these factors stimulate antibody synthesis in T cell-depleted spleen cultures. In the presence of Con A, these factors stimulate mitogenic responses in thymocyte cultures where the cell density is too low to support mitogenic responses to Con A alone. Both culture systems have been developed as quantitative assays for the activities of the Con A factors. Purification of the biologic activities by salt precipitation, gel filtration, ion-exchange chromatography, and isoelectric focusing (IEF) establish that the molecules responsible for effects on B and T lymphocytes in these assay systems are identical. The quantitative assays performed reveal that these factors are active in each culture system at concentrations below 10^-9 M. The IEF-purified factors also stimulate the specific generation of cytotoxic lymphocytes to allogeneic tumor cells in culture. These experiments reveal a requirement for helper T cells in the induction of antibody synthesis in B lymphocytes, mitogenic responses to Con A in T lymphocytes, and cytotoxic T cell precursors to effector cells. Each assay system may be a measure of the induction of antigen-sensitive lymphocytes, suggesting that B and T lymphocytes respond to similar, if not identical, helper T cell activity.

Footnotes

¹ J. W. is supported by a Research Career Development Award (AI-00182) and a Grant (AI-13383) from the National Institute of Allergy and Infectious Diseases, and Grant 1-469 from the National Foundation.

² J. S. and V. P. are supported by the National Cancer Institute of Canada.