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. 2014 Dec 8;12(6):708–718. doi: 10.1038/cmi.2014.118

Figure 2.

Figure 2

The proliferation and activation of naive CD4+ T cells is markedly inhibited by the CD8+CD28− T cells educated by MSCs. The T-cell proliferative response was evaluated by the CFSE dilution of the CFSE-labeled CD4+ T cells in the presence of anti-CD3 (200 ng/ml) and anti-CD28 (1 μg/ml). The CD8+CD28− T cells and MSCs-pre-treated CD8+CD28− T cells were added respectively at the ratio of 1∶1, and their inhibitory effects on T-cell proliferation were analyzed (a, b). The CD8+CD28− T cells and MSCs-pre-treated CD8+CD28− T cells were cocultured with naive CD4+ T cells at a ratio of 1∶1 in the presence of a stimulus for 24 h, and their effects on T-cell activation were determined by the expression of CD69 (c, d) and CD25 (e, f) on CD4+ T cells. The gray histograms correspond to the isotype controls. The bar graphs indicate the means±s.d. Statistically significant differences are indicated as follows: *P<0.05 and **P<0.01, n=3. CFSE, 5-(and-6)-carboxyfluorescein diacetate succinimidyl ester; MSC, mesenchymal stem cell.