Analysis of thymic stromal cell subpopulations grown in vitro on extracellular matrix in defined medium - V. Proliferation regulating activities in supernatants of human thymic epithelial cell cultures

In previous reports in this series, we described the growth conditions, morphology and supernatant activities of human thymic epithelial cell cultures. The human thymic epithelial cell supernatant (HTES) contained IL-6, G-CSF and M-CSF activities and exhibited a strong enhancing effect on thymocyte proliferative response to mitogens, which was identified as IL-6 related. The responding thymocyte population was apparently identified as PNA^-, mature T cells. In order to simplify further analysis of HTES activities, we selected to use a well-defined mature murine T cell clone which has a Th2 phenotype (8-5 clone). HTES induced 8-5 cell proliferation without the presence of antigen, antigen presenting cells (APC) or mitogens. This enhancing effect of HTES was completely blocked with anti hIL-6 antibody but could not be reproduced by rhIL-6 alone. Hence, IL-6 is a necessary but insufficient factor in mediating this effect. HTES induced proliferation was accompanied by endogenous IL-4 secretion from 8-5 cells. Furthermore, the proliferation was blocked by anti mIL-4 antibody, implicating IL-4 as an autocrine growth factor in this system. HTES increased also the expression of IL-2 receptor. In addition, rhIL-2 and rmIL-4 each had a synergistic effect on the proliferative response of 8-5 cells to HTES. A similar synergistic activity was demonstrated when rhIL-6 was used instead of HTES, suggesting that IL-6 regulates some of HTES activities. Our findings indicate that HTES activities, of which IL-6 is only part, are mediated via the induction of autocrine growth factors and by the regulation of T cell growth factor receptor expression.