Figure 4
Effect of anti-TIM-2 mAb treatment on antigen-specific T cell proliferation and cytokine production. (a) DBA/1 mice were immunized with type II collagen (CII)/complete Freund's adjuvant (CFA) on day 0 and CII/incomplete Freund's adjuvant (IFA) on day 21 and treated with RMT2-14, RMT2-25, RMT2-26, or control IgG from day 0 to day 42. Draining lymph node (LN) cells from 19 mice were isolated and pooled in each group at day 45 and cultured with the indicated concentrations of denatured CII (dCII). For estimating proliferation, 0.5 μCi 3H-thymidine was added during the last eight hours of a 96-hour culture. Production of IFN-γ and IL-17 in the culture supernatants at 120 hours was determined by ELISA. (b) DBA/1 mice were immunized with ovalbumin (OVA)/CFA on day 0 and treated with RMT2-14 or control IgG on days 0, 2, and 4. Draining LN cells from five mice were isolated and pooled in each group on day 7 and cultured with the indicated concentrations of OVA. For estimating proliferation, 0.5 μCi 3H-thymidine was added during the last six hours of a 72-hour culture. Production of IFN-γ and IL-17 in the culture supernatants at 72 hours was determined by ELISA. Results are expressed as the mean ± standard deviation of triplicate samples. Similar results were obtained in three independent experiments.