Fig. 7. Nod2 controls Th17 immunity in naïve mice and Blau syndrome.

a Nod2 expression in IRBP_1–20–stimulated CD4⁺ T cells of naïve, adjuvant-only, or immunized WT mice relative to unstimulated CD4⁺ T cells from naïve WT mice was measured by qPCR. Controls include IFNγ-stimulated WT BMDM and Nod2^−/− BMDM cells. Data are representative of 2 independent experiments, each with n = 5 spleens pooled/genotype. b–c Rag2^−/− hosts reconstituted with CD4⁺ T cells from naïve R161M or Nod2^−/−R161M mice were assessed for uveitis 7 days post-transfer. Data are combined from 3 independent experiments with n = 12 mice/group. b Clinical uveitis was scored, *p = 0.0500. c Representative fundus and histopathology photographs. d–g T cell responses in naïve WT and Nod2^−/− (C57BL/6 J) mice. d Naïve CD4⁺ T cells (CD62L^hiCD44^-) from WT and Nod2^−/− mice were differentiated under Th17-polarizing conditions and frequency of IL-17-producing CD4⁺ T cells (CD4⁺CD8^-Thy1.2⁺IL-17⁺) was determined by flow cytometry. Data are combined from 2 independent experiments for n = 8 mice/genotype. e Frequency of memory (CD62L^-CD44⁺) CD4⁺ T cells from naïve WT or Nod2^−/− mice expressing indicated factors was determined by flow cytometry 5 days post-stimulation with anti-CD3/anti-CD28 mAb. Data is representative of 3 independently performed experiments, each with n = 4 mice/genotype; *p = 0.0286 for all indicated comparisons. f–g Memory CD4⁺ T cells from naive WT or Nod2^−/− mice stimulated 16 h with anti-CD3/anti-CD28 mAb were analyzed by multiplex-qPCR. Transcripts are relative to unstimulated control memory CD4⁺ T cells of WT mice (horizontal line), with n = 3 mice/condition. h–i PBMCs of two healthy controls or two Blau syndrome patients were stimulated with anti-CD3/anti-CD28 mAb for 5 days then with PMA/ionomycin for 4 h. Experiment was repeated twice. h Frequency of IL-17-producing CD4⁺ T cells (CD3⁺CD4⁺CD19^-IL-17⁺)(*p = 0.0055) and representative contour plot. i Representative histogram showing MFI of CCR7 by CD4⁺ T cells (CCR7⁺CD3⁺CD4⁺CD19^-). Data are box-whisker plots showing median with 25–75th percentile and min–max range (b, d), box-whisker plots showing median with 10–90th percentile (e), or floating bars showing median and min–max range (f, g). Dots represent individual values (b, h). Statistical differences were calculated using two-tailed Mann–Whitney U test (d, e) or unpaired, two-tailed Student’s t-test (b, f–h). Source data are provided as a Source Data file.⁺.