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TH17 differentiation from naïve CD4 T cells
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Lymphocytes from spleen and dLN were pooled and sorted using a Reflection (i-Cyt). Sorted naïve CD4+ T cells (CD4+CD25−CD62L+CD44−) were activated in vitro for 4–5 days with 2 μg/ml anti-CD3 (2C11; Bio X Cell), 2 μg/ml anti-CD28 (37.51; Bio X Cell), human TGF-β1 (2 ng/ml, R&D) and mouse IL-6 (20 ng/ml, BD Biosciences). Cytokine expression was assessed after PMA/ionomycin stimulation as described above.
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