Fig. 6. Targeting AHR or IL-22RA1 had an impact on thymus regeneration, T-cell reconstitution and pathogenesis of murine cGVHD following allo-HCT.
In the cGVHD models, BALB/c mice were used as donors. Wild type C57BL/6 (a), B6-Il22ra1fl/fl and B6-Il22ra1fl/fl;Foxn1-cre mice (b) were recipients. Wild type C57BL/6 recipients were treated by vehicle or FICZ thrice weekly for 2 weeks. a, b Survival (n = 15 in each group) and cGVHD score (Day 60) of recipient mice were monitored. c Histological analyses (Day 90): H&E staining, Masson staining and collagen immunofluorescence staining (n = 4). Ashcroft score was determined by inflammation and fibrosis. Area of collagenous and fluorescence intensity was analyzed using ImageJ. Scale bar: 50 μm. d, e At day 90, thymus cells were counted as described (n = 7 or 5). f At day 90, blood samples were collected and flow cytometry was used to analyze T-cell subpopulations (n = 7 or 5). g At day 90, T cells were isolated from spleen cells of recipients. Lymphocytes were isolated from spleen cells of normal C57BL/6 mice. The isolated recipients’ T cells were cultured alone (control) or co-cultured (MLR) with normal lymphocytes (1:1) for 72 h. Cell viability was detected by CCK-8 assay (n = 7 or 5). Survival is compared using log-rank test. Data are mean ± SD, compared using one-way ANOVA test or Student’s t test. *, p < 0.05; **, p < 0.01; n.s., not significant.