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Activated myelin specific T cells are thought to mediate inflammatory tissue damage in multiple sclerosis (MS). Applying a large panel of myelin antigens, we demonstrate the direct ex vivo detection of viable IFN γ/TNF αproducing CD4 +/CD69+T cells 6 hours after antigenic challenge, by intracellular flow cytome try in 3/33 MS patients and 2/26 healthy controls with calculated frequencies of (mean ±.SEM): 0.031%±0.002%versus 0.037%±0.029%. By comparison, the rece ntly developed IL 7 modified proliferation assay revealed i) a higher number of individuals showing myelin reactivity (17/37 MS patients and 12/24 healthy indi viduals) and ii) a significant difference in the response to myelin basic protei n (MBP) between the two groups in a longitudinal analysis, indicating a higher a ctivity of myelin specific T cells in MS patients. Our data provide new perspec tives in detecting pathogenetically relevant T cells, but clearly demonstrate th e different conclusions which must be drawn from various approaches concerning t he quantification of autoreactive T cells.
Applying a large panel of myelin antigens, we demonstrate the direct ex vivo detection of viable IFNγ / TNFαproducing CD4 + / CD69 + T cells 6 hours after antigenic challenge, by intracellular flow cytome try in 3/33 MS patients and 2/26 healthy controls with calculated frequencies of (mean ± .SEM): 0.031% ± 0.002% vs. 0.037% ± 0.029%. By comparison, the recently developed IL 7 modified proliferation assay i) a higher number of individuals showing myelin reactivity (17/37 MS patients and 12/24 healthy indiiduals) and ii) a significant difference in the response to myelin basic protei n (MBP) between the two groups in a longitudinal analysis, indicating a higher a ctivity of myelin specific T cells in MS patients. Our data provide new perspec tives in detecting pathogenetically relevant T cells, but clearly demonstrated th e different conclusions which must be drawn f rom various features concerning t he quantification of autoreactive T cells.