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. 2020 Sep 4;11:2071. doi: 10.3389/fimmu.2020.02071

FIGURE 3.

FIGURE 3

Adoptive transfer of HEL protein-activated 3A9 CD4+ T cells but not DN T cells induces severe EAU in IRBP:HEL sTg mice. (A) Antigen-specific CD4+ T cells and DN T cells were sorted from P50-70 3A9 TCR mice, labeled with CFSE proliferation dye and incubated with HEL protein (1 μM) for 72 h. Fundus images of sTg mice were taken, and retinal appearance clinically scored before adoptive transfer (Tx) at baseline (0; P20), and then at d5 (P25), d8 (P28) and on d10 (P30) after Tx (n = 6/time point). 2 × 106 CD4+ cells induced EAU within 8 days while 5 × 106 CD4+ cells induced severe EAU within 5 days of Tx. Adoptive transfer of DN CD3+ cells did not induce EAU despite being activated by HEL antigen even with doses of 10 × 106 cells (data not shown). Data were analyzed for each time point using one-way ANOVA and Tukey’s Multiple Comparison post hoc Test with *p < 0.05, ***p < 0.001 on a 95% level of confidence. (B) Representative images of heamatoxylin and eosin (H&E) stained sections from adult (P45-P60) sTg IRBP:HEL mouse eyes, 8 days following Tx of HEL-activated CD4+ T cells from 3A9 TCR mice of the same age. Adoptive transfer induced a severe, multifocal panophthalmitis (17) (a) Cyclitis with “plastic” infiltrate in the vitreous and cellular precipitates on posterior lens surface (arrowhead). (b) Central retinal vasculitis and optic nerve destruction with granuloma plus some surrounding intact retinal tissue remaining (boxes). (c) Tertiary lymphoid organ (TLO; arrowhead) surrounded by edematous and necrotic retinal tissue (pale staining region; asterisk) (d) Inflammation of the ciliary body (bracket) (cyclitis) and posterior chamber (arrowheads) with plastic exudate in the anterior chamber. (e) Plastic deposit in the anterior chamber with keratic precipitates adherent to the corneal endothelium (bracket). (f) Massive cell infiltration in iris (iritis) with two granulomas (arrowheads). R, retina; CH, choroid; S, sclera; C, cornea; ON, optic nerve; AC, anterior chamber; L, lens; I, iris; PS, posterior segment; CB, ciliary body; Co, conjunctiva.