Fig. 5.

Immunomodulatory effects of human bone marrow MSCs on aberrant T subsets and cytokines profile. a, b NKT-PBMCs were co-cultured with human MSCs or human fibroblasts at a 20:1 ratio of NKT-PBMCs to human MSCs or NHLF prior to cytokines test in the supernatants (a) and flow cytometric analysis (b) for each group. Triplicate wells were prepared for each group. a TNF-α, IFN-γ, TGF-β1, and IP-10 levels in the supernatants of NKT-PBMCs, MSCs, and NKT-PBMCs co-cultured with human bone marrow MSCs or NHLF. ** Significantly different from the NKT-PBMCs group, P < 0.01. † P < 0.05, †† P < 0.01, compared to MSCs or NKT-PBMCs co-cultured with NHLF. Data represent the means ± SD from three independent experiments. b Flow cytometric analysis of CD3⁺ CD56⁺ cells, CD3⁺ CD8⁺ cells, CD3⁺ CD4⁺ cells gating on CD45⁺ cells, and CD25⁺ CD127^(Low/-) Treg cells gating on CD4⁺ cells, of either NKT-PBMCs (NKT-PBMCs) or NKT- PBMCs co-cultured with human bone MSCs (NKT-PBMCs/MSC, or co-cultured with NHLF (NKT-PBMCs/NHLF). *P < 0.05 for comparisons between NKT-PBMCs/MSC and NKT-PBMCs/NHLF or NKT-PBMCs. Data represent the means ± SD from three independent experiments. c CD25⁺ FOXP3⁺ Treg cells gating on CD4⁺ cells in the PBMCs of healthy controls and IPF patients (n = 12) before and after being co-cultured with MSCs or human fibroblasts. Data represent the means ± SD. *P < 0.05. MSCs mesenchymal stem cells, NKT natural killer T cells, PBMCs peripheral blood mononuclear cells, NHLF normal human lung fibroblasts, TNF-α tumor necrosis factor-α, IFN-γ interferon γ, TGF-β transforming growth factor-β, IP-10 interferon γ-induced protein 10