Figure 3.

PD-L1- and CD86-functionalized MSCs upregulate PD-1 and CTLA-4 pathways in myelin-specific T cells, downregulate T cell activation and promote the development of induced regulatory T cells in vitro. a,b) PD-1 (a), and CTLA-4 (b) expressions of myelin-specific 2D2 T cells after incubated with different types of PD-L1 FcIg- and/or CD86 FcIg-functionalized MSCs for 48 h, as determined by FACS assay. Cells were initially gated at CD3⁺ cells. (n = 4) c,d) ELISA analysis of INF-gamma (c) and IL-17A (d) secreted from 2D2 CD4⁺ T cells after incubated with different functionalized MSCs. Supernatants were collected 48 h post-incubation for the ELISA analysis. (n = 4) e) Quantification of IL-10⁺ and FoxP3⁺ population in 2D2 CD4⁺ T cells after incubated with different functionalized MSCs for 48 h via FACS. Cells were initially gated at CD3⁺ cells. (n = 3) Statistical significance between two control/experimental groups were accessed by two-way ANOVA with Tukey correction. Data represent mean ± s.e.m.