(A) WT and IL-10-/- CD4+ T cells isolated using MACS were stimulated at day 0 as before. After 24 hours, cells were washed to remove soluble agonists, and then allowed to incubate for an additional 3 days. Conditioned media supernatants, which were analyzed for IL-10 by ELISA (B), were then transferred to fresh recipient CD4+ T cells stimulated by αCD3 on day 4. On day 7, the IFNγ response of the recipient cells was quantified. Normalized to the response without transferred supernatant (100%; dotted line) and subtracting IFNγ concentrations in the donor supernatants, we found that supernatants from TCR/TLR2 co-stimulated WT T cells inhibited the recipient IFNγ response by 50%, whereas supernatants from TLR2-only stimulated T cells augmented recipient IFNγ production by over 2 fold (C). In contrast, supernatants generated from all stimulated IL-10-/- donor T cells promoted IFNγ by the recipients by nearly 2 fold (D). All data are n≥3, unless otherwise noted; *p<0.05; #p>0.05.