Helper cells in murine thymus for the generation of primary cytolytic T lymphocyte (CTL) responses in vitro.

Thymocytes used as responding cells in a mixed leukocyte culture with gamma-irradiated splenic stimulating cells generated highly significant proliferative and cytolytic responses when responding and stimulating cells differed by the entire H-2 complex or at H-2K plus H-21 regions. On the other hand, when the difference was only an H-2K region, very little, if any, proliferative response was detectable and no cytolytic response was found. Because it has been claimed by others that thymocytes do not include cells needed to provide the help required for the generation of a cytolytic response and yet we have found a highly significant response in thymocytes, we have analyzed, in 2 systems, the possible role of back-response by stimulating spleen cells in the generation of cytolytic activity by responding thymocytes. In the first system, spleen cells were depleted of T lymphocytes by treatment with anti-Thy-1.2 antiserum plus complement. In the second system, F1 hybrid mouse spleen cells were used to stimulate parental responding thymocytes. In both cases, no back-response as measured by cell proliferation in mixed leukocyte culture was detected, while both types of spleen cells still stimulate significant cell proliferation and cytolytic activity from responding thymocytes, as well as from responding spleen cells. Furthermore, it was found that in the presence of stimulating spleen cells (either T lymphocyte-depleted or not) that differ from the responding thymocytes at the entire H-2 complex, thymocytes were capable of generating a significant and specific cytolytic response to H-2K region different stimulating spleen cells (either T lymphocyte-depleted or not). These results are compatible with the suggestion that there are sufficient numbers of helper cells in the unfractionated thymocyte population for the generation of primary cytolytic T lymphocyte responses in vitro.