Figure 3

SHED interplay with T-lymphocytes. (A, B) Under the anti-CD3 and CD28 antibody along with TGFβ1 and IL-2 stimulation, SHED showed a significant effect in reducing Th17 cell levels as seen in BMMSCs (A), however, SHED exhibited a significant capacity of inhibiting IL17 levels than BMMSCs (B) (n = 3, *P < 0.05,***P < 0.001). (C) PBMNCs activated by anti-CD3 antibody (@CD3Ab, 1 μg/ml) were capable of inducing significant SHED and BMMSC death as shown by toulidin blue staining. When cells were cultured in an indirect co-culture system using Transwell, activated slpenocytes they failed to induce SHED and BMMSC death. Neutralization with anti-FasL antibody (@FasLAb, 1 μg/ml) blocked PBMNC-induced SHED and BMMSC death. Representative of n = 3. (D) SHED express a higher level of Fas in comparison to that in BMMSCs by immunoblotting. Three independent experiments showed similar results. Representative of n = 3. (E) SHED death caused by active PBMNCs is through an apoptotic pathway according to the TUNEL staining. The SHED death rate was similar to BMMSCs. The percentage of TUNEL-positive (TUNEL+) nuclei was indicated to the total number of MSCs and averaged from five replicated cultures (***P < 0.005).