Blood -- Regulation of Th17 cell differentiation and EAE induction by MAP3K NIK

Blood, Vol. 113, Issue 26, 6603-6610, June 25, 2009

Regulation of Th17 cell differentiation and EAE induction by MAP3K NIK
Blood Jin et al. 113: 6603

Supplemental materials for: Jin et al

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Figure S1. NIK is essential for EAE induction (JPG, 67.1 KB) -
Age-matched wildtype (NIK^+∕+) and NIK knockout (NIK^−∕−) mice (5^−∕− and 7^+∕+) were immunized with MOG_35–55 peptide on day 0 and day 8 and monitored daily for EAE symptoms, which are presented as percent of incidence.
Figure S2. NIK regulates the strength of TCR and LIGHT costimulatory signals involved in Th17 differentiation (JPG, 549 KB) -
(A) Naïve CD4 T cells isolated from WT (NIK^+∕+) mice were stimulated for 48 h with plate-bound anti-CD3 and anti-CD28 (1 µg/ml) under Th0 or Th17 conditions. Relative expression of the indicated genes was analyzed by real-time RT-PCR and normalized to the expression of actin mRNA. The gene expression level induced under Th17 conditions was presented as fold over that induced under Th0 conditions (set to 1). (B) WT naïve CD4 T cells were stimulated under Th1, Th2, or Th17 conditions for 48 h and subject to real-time PCR analysis as described in A. Relative HVEM mRNA level was presented as fold over the value obtained under Th1 conditions. (C) and (D) Naïve CD4 T cells isolated from NIK^+∕+ and NIK^−∕− mice were stimulated with the indicated amounts of anti-CD3/CD28 under Th17-polarizing conditions, either in the absence (−) or presence (+) of recombinant LIGHT (500 ng/ml). Cells stimulated under Th0 conditions (with 1 µg/ml anti-CD3 and anti-CD28) were included as controls. After 72 hr, the percentage of Th17 cells was analyzed by ICS based on the production of IL-17.