Mice undergoing short-term treatment with BIO 5192 initiated at the peak of acute disease display enhanced proliferation and IFN-γ secretion in response to the priming PLP139-151 epitope and augmented epitope spreading. Spleen cells from 5 mice per group were harvested after 22 treatments (day 39 after priming) from the mice in Figure 2. (A) Viable spleen cells (5 × 105/well) were cultured with indicated concentrations of PLP139-151 for 4 days and proliferation was assessed by incorporation of 3H-TdR. Results are expressed as ΔCPM (media only backgrounds subtracted). Stimulation indices are shown above the relevant bars. (B) Supernatants from the above culture in panel A were harvested at 48 and 72 hours and analyzed for IFN-γ secretion in response to PLP139-151 by ELISA as described in “Materials and methods.” (C) Proliferative responses to the relapse-associated PLP178-191 epitope were also assessed from splenic cultures. (D) Supernatants from the above culture in panel C were harvested at 72 hours and analyzed for IFN-γ secretion in response to PLP178-191 and MBP 84-104 by ELISA as described in “Materials and methods.” Error bars indicate SEM.
