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. 2017 Jan 11;12(1):e0169362. doi: 10.1371/journal.pone.0169362

Fig 5. IFNγ-induced expression of IDO by MSCs is necessary to inhibit Vδ2+ cell proliferation.

Fig 5

(A) Lentiviral-mediated shRNA transduction of MSCs was used to knock down IDO (MSC-IDOi). Quantification of IDO mRNA by Real-Time qPCR was done in MSC-pLV, MSC-IFNγRi and MSC-IDOi. Relative expression was normalized to MSC-transduced with the empty vector (B) Representative flow cytometric analysis of activated Vδ2+ cells after five days of co-culture with transduced MSCs. Vδ2+ cells expand more in the presence of MSC-IDOi compared to MSC-pLV. (C) Specific silencing of IDO augments the number of Vδ2+ cells after five days of co-culture. Results show the means ± S.D. of triplicate samples. **P ≤ 0.01. (D) Representative proliferation analysis of CFSE-labeled Vδ2+ cells performed at day five of co-culture with either MSC-IDOi (white) or empty vector (grey). Proliferation index of Vδ2+ cells in co-culture with MSCs silenced with shRNA for IDO is higher compared to MSC transduced with the empty vector (right panel), similar to the results obtained with MSC-IFNγRi. Results show the means ± S.D. of triplicate samples. *P ≤ 0.05. (E) Co-culture of Vδ2+ cells and MSC-IDOi gives rise to more intracellular IFNγ production of Vδ2+ cells compared to a co-culture with MSC-pLV after 12h. Results show the means ± S.D. of triplicate samples. **P ≤ 0.01. Representative flow cytometric analysis of IFNγ production in Vδ2+ cells co-cultured with MSC-IDOi after 12h of activation (right panel).