Nuclear Receptor NR4A2 Orchestrates Th17 Cell-Mediated Autoimmune Inflammation via IL-21 Signalling
Figure 1
Autoimmune induction of NR4A2 in CD4+ T cells is associated with IL-17-secreting T cells.
EAE or EAU was induced in C57BL/6 mice by immunization with MOG35–55 or IRBP1–20 peptide in CFA, respectively. CD4+ T cells were purified from spleen, blood, or target organ (CNS or retina) on the indicated days and RNA was isolated. A and B: NR4A2 expression was quantified by real time PCR relative to GAPDH for T cells from EAE (A) or EAU (B). Timepoints correspond to a minimum of 5 animals and data are representative of 3 independent experiments. CD4+ T cells from mice with EAE were restimulated with PMA/ionomycin for 3 hours and 4 populations of cytokine secreting cells (IL-17+IFN-γ-, IL-17+IFN-γ+, IL-17-IFN-γ+, and IL-17-IFN-γ-) were sorted by flow cytometry using IFN-γ and IL-17 cytokine secretion assay kits. C: NR4A2 expression by populations of cytokine-secreting CD4+ T cells was quantified by real time PCR at day 15 post-EAE induction for lymph nodes (LN) and CNS-infiltrating cells (CNS), and day 25 for blood T cells. D and E: NR4A2 expression by IL-17+IFN-γ− or IL-17+IFN-γ+ CNS-infiltrating T cells (D) or blood T cells (E) was measured by real time PCR at a range of timepoints. Data are representative of 2 independent experiments. F: Th1-mediated diabetes was induced in C57BL/6 mice by 5 daily low dose STZ treatments. Other groups of C57BL/6 mice were immunized with peptides in CFA plus PTX either OVA323–339 (OVA/CFA) or MOG35–55 (MOG/CFA). On day 22, NR4A2 expression was assessed by real time PCR amongst CD4+ T cells from spleen and leukocytes isolated from the relevant target organ (ND, OVA/CFA; CNS, EAE; pancreas, STZ). Timepoints correspond to a minimum of 5 animals and data are representative of 2 independent experiments.