Administration of anatabine inhibits pro-inflammatory cytokines but does not affect the titer of circulating anti-MOG antibodies.

A) Average amount of anti-MOG antibodies measured by ELISA in the serum of control non-immunized mice, EAE placebo and EAE anatabine treated mice at day 16 following the MOG immunization. ANOVA reveals a significant main effect of the MOG immunization on the titer of anti-MOG antibodies (P<0.001) but no significant main effect of anatabine (P = 0.617). B) Anatabine inhibits the production of pro-inflammatory cytokines in the serum of EAE mice. A significant elevation of TNF-α (Mann-Whitney U = 16.5, Z = −2.22, P = 0.026), IL-1β (Mann-Whitney U = 0, Z = −3.506, P<0.001), IL-6 (Mann-Whitney U = 3, Z = −3.271, P<0.001), IFN-γ (Mann-Whitney U = 16.5, Z = −2.219, P = 0.026) and IL-17 (Mann-Whitney U = 7, Z = −2.958, P = 0.003) was observed in the serum of EAE mice (placebo) compared to control non-immunized mice. Anatabine significantly prevented the production of IL-1β (Mann-Whitney U = 44, Z = −2.666, P = 0.008), IL-6 (Mann-Whitney U = 59, Z = −2.008, P = 0.045) and IL-17 (Mann-Whitney U = 47, Z = −2.531, P = 0.011) in the serum of EAE mice. C) Anatabine suppresses pro-inflammatory cytokine production in the spleen of EAE mice. A statistically significant elevation of TNF-α (Mann-Whitney U = 3, Z = −3.270, P = 0.001) and IL-1β (Mann-Whitney U = 0, Z = −3.503, P<0.001) was observed in the spleen of EAE placebo mice compared to control non-immunized mice. A significant decrease in TNF-α (Mann-Whitney U = 22, Z = −3.622, P<0.001), IL-1β (Mann-Whitney U = 9, Z = −4.19, P<0.001) and IL-17 (Mann-Whitney U = 55, Z = −1.976, P<0.05) was observed in the spleen of anatabine treated EAE mice compared to placebo EAE mice. (* P<0.05 for EAE placebo mice vs control non-immunized mice; ¤ P<0.05 for EAE placebo mice vs EAE anatabine treated mice).