Generation of T Helper Cells in Vitro

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Abstract
In the thymocyte-macrophage (T-Mø) culture system for the in vitro induction of murine T helper cells, cortisone-resistant, nylon wool-purified thymocytes (CRNPT) are cultured with macrophages from antigen-primed donors. The particulate antigens, sheep erythrocytes (SRBC) or vaccine prepared from cell walls of Group A Streptococcus (SAV), were used to prime T-Mø cultures. The native CRNPT population contained Ly1, Ly23, and Ly123 cells in the proportions 45:3:46, but after 4 days of culture with antigen-pulsed macrophages the proportions were 88:2:2. The Ly1 T cell populations selected from such cultures by treatment with anti-Ly2 serum plus complement exhibited antigen-specific helper or suppressor functions in vitro, depending on the mode of the PFC assay: when assayed for their effect on T celldepleted spleen populations, helper function was manifest; when assayed with intact spleen cell populations, suppression was manifest. Our data do not indicate whether these two functions are mediated by the same Ly1 cells or by two subpopulations of Ly1 cells. The generation of Ly1 helper cells from CRNP thymocytes in 4-day T-Mø culture required the initial presence of the Ly123 members of this population. Manifestation of suppression by the same Ly1 population required the presence of Ly123 cells in the splenic populations used for PFC assay.

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