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. 2020 Oct 26;11:5406. doi: 10.1038/s41467-020-18961-0

Fig. 2. Nod2 influences retinal antigen-specific cellular responses.

Fig. 2

a The proportion of ocular CD45+ cells of WT and Nod2−/− mice 20 days post-immunization was determined by flow cytometry. Dashed line denotes baseline level for adjuvant control eyes. b Frequency of macrophages (CD45+CD11b+F4/80+), neutrophils (CD45+CD11b+GR-1+), dendritic cells (CD45+CD11c+), and T cells (CD45+CD3+CD4+) are summarized as stacked bars. c Live T cell populations of gated CD45+ cells. Data (a–c) are representative of 4 independent experiments each with n = 4 mice/group; *p = 0.0286 (a, c). d Uveitis was evaluated in immunized WT or Nod2−/− mice that were depleted of CD4+ T cells with α-CD4 mAb or Isotype control Ab (IC Ab). Data are representative of 2 independent experiments with n = 12 mice/group; ***p < 0.0001 (WT IC vs. WT Ab, Nod2−/− IC vs. Nod2−/− Ab, WT IC vs. Nod2−/− IC), and *p = 0.0373 (WT Ab vs. Nod2−/− Ab). e–g Pinealitis was assessed in WT and Nod2−/− mice 14 days post-immunization. e Pinealitis scores from histopathological evaluation, and (f) representative H&E-stained sections (circle denotes dense lymphocytic infiltration). Scores in e were combined from 2 independent experiments (shown left to right with n = 4, 6, 4, 5); *p = 0.0433 (WT IRBP vs. Nod2−/− IRBP), *p = 0.0159 (Nod2−/− adjuvant vs. Nod2−/− IRBP). g Frequency of CD4+ and CD8+ T cells in pineal glands was determined by flow cytometry. Data are combined from 3 independent experiments, each having 5 pooled pineal glands/genotype/treatment. For CD4+ cells: *p = 0.0111 (WT IRBP vs. Nod2−/− IRBP) and *p = 0.0081 (WT adjuvant vs. Nod2−/− adjuvant). For CD8+ cells: *p = 0.0122 (WT adjuvant vs. Nod2−/− adjuvant). h BM chimeric mice were assessed for EAU by histopathology 20 days post-immunization. Data are combined from 3 independent experiments (shown left to right with n = 31,31,30,28); **p = 0.0030 and ***p < 0.0001. Data are box-whisker plots showing median, 25th–75th percentile and min–max range (a, c–e, h), floating bars showing median with min–max range (g), or mean (b). Data were analyzed with two-tailed Mann–Whitney U test (a–e, h) or unpaired, two-tailed Student’s t-test (g). Source data are provided as a Source Data file.