The inductive requirements for the primary in vitro generation of delayed-type hypersensitivity response to influenza virus in mice

Effector T cells (Td) which mediate delayed-type hypersensitivity reactions to influenza A virus can be generated in tissue culture using normal mouse spleen cells as the responder population. Addition of helper T cells enhances but is not essential for the production of Td cells. Both Ly 1 positive, I region restricted and Ly 2,3 positive, K,D region restricted effector cells are generated. Treating the responder cell population with anti-Ly 1 or anti-Ly 2,3 antibodies and complement prevented the generation of both classes of effector T cell, suggesting that the precursor Td cells are Ly 1,2,3 positive. Effector cells which are specific for the homologous virus or cross-reactive within the A strains of influenza virus are produced, as has been found previously in in vivo experiments. Depleting the cell population of phagocytic and plastic adherent cells resulted in a failure to produce Td cells, which showed a requirement for macrophage-like cells as accessory cells in the primary in vitro generation of Td cells. A variety of cells, such as peritoneal exudate cells, mitogen stimulated blasts or L929 fibroblast cells could serve as stimulator cells. Only Ly 2,3 positive, K,D region-restricted Td cells were produced when L929 cells were used as they lack I region-coded surface antigens. The I region-restricted DTH response was mapped to the IA sub-region of the H-2 gene complex.