Fig 1: Increased Reactivity of Memory CD4+ T Cells against HLA-DR-SPs in MS Patients(A) Responses of CD45RA- PBMCs of HLA-DR15+ HDs (n = 8) and MS patients (RRMS, n = 4; RRMS_NAT, n = 10) against the five most common HLA-DR-SPs alone or as pool. 10–15 replicate wells were tested for proliferation, and responses are depicted as the stimulatory index (SI). Individual wells are represented by dots for HDs (left) or MS patients (right). The purple dotted line indicates SI = 2, and SI = 2 was considered positive. Responses to control CEF II peptides are shown at the bottom.(B) Proliferations of CD45RA- PBMCs from HDs and MS patients after stimulation with pooled HLA-DR-SPs in the presence of a blocking anti-HLA-DR Ab.(C) CD45RA- PBMCs from HLA-DR15+ MS patients were labeled with carboxyfluorescein succinimidyl ester (CFSE) and stimulated with pooled HLA-DR-SPs. After 7 days, cells were analyzed, and proportions of memory CD4+ T cells in the proliferating (CFSEdim) and highly proliferating (CFSElow) compartments are shown in the pie charts.(D) Blood CD4+ T cells and monocytes were purified from PBMCs of HLA-DR15+ MS patients and co-cultured with pooled HLA-DR-SPs for 7 days. Proliferation of naive CD4+ T cells (CD45RA+) and memory CD4+ T cells (CD45RA-) was detected.(E) Th1/Th2/Th17-related cytokines in supernatants of CD45RA- PBMCs after stimulation with pooled HLA-DR-SPs.(F) IFN-?, IL-17A, and IL-17F in supernatants of co-cultured blood CD4+ T cells and monocytes after stimulation with pooled HLA-DR-SPs.(G) Expression levels of CCR6 and CXCR3 on non-proliferating (CFSEhigh) and proliferating (CFSEdim) memory CD4+ T cells after stimulation with pooled HLA-DR-SPs for 7 days.Data are expressed as mean ± SEM, and p values were determined by unpaired t test. See also Figure S4 and Tables S1 and S5.