Figure S2.

IFN-γ Secretion Is Strongly Increased during AP in RRMS (REM) but Is Not the Cause of Increased AP, Related to Figures 1 and 2 and Table S1

(A) A second cohort of HD (n = 14; for PHA n = 10) and REM (n = 14; for PHA n = 12) was used to assess AP (unstimulated) and reactivities to conventional B- (α-IgM) or T cell stimulation with polyclonal/broad (MLR, PHA) or antigen-specific (TTx) activation using CFSE-labeling and subsequent co-culture for 7 days in serum-free medium. Frequency of all proliferating (CFSE^dim) cells for each HD and REM under the different conditions is depicted (mean ± SEM; Mann-Whitney U test).

(B) Secretion of Th1 (IL-2, TNF, IFN-γ), Th2 (IL-4, IL-5, IL-13) and Th17 (IL-17A, IL-21, IL-22) cytokines in supernatants collected after 7 days of AP (no stimulus), conventional B- (α-IgM) or T cell stimulation with polyclonal (MLR) or antigen-specific (TTx) activation from HD (n = 14) and REM (n = 14; for TTx n = 13) (mean ± SEM; Mann-Whitney U test).

(C) Levels of IFN-γ and GM-CSF secretion in HLA-DR15^-and DR15⁺ HD (DR15- n = 20 and DR15+ n = 12) and REM (DR15^- n = 15 and DR15⁺ n = 17; whiskers: min - max). Cytokine measurement was performed with bead-based immunoassay and GM-CSF by ELISA.

(D) Frequency of CFSE^dim cell population (gray bars) in the presence of blocking IFN-γ, blocking GM-CSF or appropriate isotype control antibodies (mean ± SEM; n = 5 REM). Cytokine neutralization (blue line) in supernatant is depicted exemplarily for one of the donors.