Production of macrophage-activating and migration-inhibition factors in vitro by serologically selected and cloned Listeria monocytogenes-specific T cells of the Lyt 1+2- phenotype

Abstract

Lyt-selected Listeria-immune T lymphocytes from peritoneal exudates and cloned T cells were cocultured with heat-killed listeriae and peritoneal macrophages from nonimmune donors. Supernatants were assayed for: activation of macrophages for tumoristatic and tumoricidal activity via macrophage-activating factors and migration-inhibition factor activity. Peptone-induced peritoneal macrophages were activated by incubation with the supernatants for 24 h. For examination of cytocidal activity, 51Cr-labeled EL4 tumor cells were subsequently added, and 51Cr release was determined. Cytostatic activity was measured by adding unlabeled EL4 tumor cells to the pretreated macrophages and determining [3H]thymidine incorporation 24 h later. Migration-inhibition factor production was examined in an agar microdroplet assay. Only Listeria-specific T cells of the phenotype Lyt 1+2- proved active in these assays, whereas T cells of the phenotype Lyt 1-2+ were not active. When T-cell clones were used, a single clone was capable of inducing macrophage-activating and migration-inhibition factor production at cell concentrations of ca. 10(3)/ml.