Fig. 4
MICCop therapy does not suppress the immune response to foreign antigens. a MICCop- (n = 8) or PBS-treated (n = 8) EAE mice were immunized with ovalbumin (OVA) on day 55 and 74 after disease induction. Additional controls comprised healthy animals with OVA (naïve + OVA, n = 4) and w/o OVA immunization (naïve w/o OVA, n = 2). Animals were sacrificed on days 107 and 111. Serum was obtained and peripheral mononuclear cells from lymph nodes (LNCs) were isolated. b Anti-OVA antibodies were detected by ELISA in sera of single animals diluted 2500 to 312,000-fold. The graph shows mean values ± standard deviation (SD). For comparison of the different groups with the naïve w/o OVA control the unpaired Student’s t test was used (***p < 0.001). c and d OVA- and PLP-specific T-cell proliferation of LNCs harvested from single animals was assessed by in vitro restimulation with OVA protein (c) and PLP139–151 (d). Proliferation was determined after 48, 72 and 96 h by [3H]-thymidine incorporation and is indicated as x-fold increase in relation to unstimulated cells (ordinate). Shown is the mean ± standard error of the mean (SEM) of every group. The differences of T-cell responses towards OVA among the groups naïve + OVA, EAE + OVA and EAE + MICcop + OVA were statistically not significant ( c) whereas the proliferative response of MICCop-treated mice upon PLP stimulation was still suppressed after OVA immunization ( d; EAE + MICCOP + OVA vs. EAE + OVA after 48 h: p < 0.01; 72 h: p < 0.05; 96 h: p < 0.01). As control, LNC proliferation against OVA of naïve mice immunized with OVA was significantly stronger than that of naïve mice without OVA treatment (96 h: p < 0.01). Two-way-ANOVA test with Bonferroni correction was used