Effect of soluble mediators on the suppressor activity of anergic T cells.
Soluble mediators were found to be not essential for the suppressor activity of anergic T cells. Alloantigen-specific anergic CD4+ T cells (1 × 105) were cocultured with an equal number of activated control T cells (1 × 105) (CTRL) and stimulated with mature DCs (1 × 104) from the same donor used during induction of anergy. A coculture of control syngeneic CD4+ T cells (1 × 105) (CTRL) with DCs served as controls. Blocking antibodies to IL-10, TGF-β, and IL-2 (100 U/mL) were added to coculture experiments. In some experiments, supernatants of anergic T cells were harvested and added to coculture of syngeneic control T cells. T-cell proliferation was measured after 3 days of incubation and an additional 16-hour pulse with3H-Tdr (37 kBq per well) by means of a liquid scintillation counter.
