Figure 7. MSC-induced CD11b^midLy6C^midLy6G^lo cells protect against EAU development in mice.

(A) The CD11b^loLy6C^loLy6G^lo, CD11b^midLy6C^midLy6G^lo, and CD11b^hiLy6C^hiLy6G^lo cells were sorted as in Figure 5A and stimulated with LPS for 18 hours. Each cell population or the vehicle (Hanks balanced salt solution [BSS]) was injected i.v. into mice immediately after EAU induction (day 0). Twenty-one days later (day 21), the mice were sacrificed and assayed. (B–D) Representative microphotographs of H&E staining, CD3 immunostaining of the retinal cross-sections, and disease score assigned by histological findings. The retinal structure, especially outer nuclear layer, including photoreceptor nuclei (arrowheads), was disorganized and infiltrated with inflammatory cells and CD3⁺ cells in the CD11b^hiLy6C^hiLy6G^lo cell–treated EAU mice. In contrast, the retinal structure was preserved and few inflammatory cells were observed in mice treated with CD11b^midLy6C^midLy6G^lo cells. Scale bar: 100 μm. (E and F) Representative flow cytometry cytograms and quantitative results for IFN-γ⁺CD4⁺ cells and IL-17⁺CD4⁺ cells in draining cervical lymph nodes (CLN). The numbers presented in cytograms (E) represent the percentage of IFN-γ⁺ or IL-17⁺ population of CD4⁺ cells, and the data shown in quantitative graphs (F) are the percentage of IFN-γ⁺CD4⁺ cells or IL-17⁺CD4⁺ cells of total CLN cells. (G) Real-time RT-PCR analysis for the proinflammatory cytokines in the eye. Shown are the values of mRNA levels relative to those in normal eyes without EAU. A dot indicates data from 1 individual animal (mean ± SD). Each biological sample was assayed in 3 technical replicates for RT-PCR. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001 by 1-way ANOVA and Tukey’s multiple-comparison test.