Mesenchymal Stem Cell-Secreted TGF-β1 Restores Treg/Th17 Skewing Induced by Lipopolysaccharide and Hypoxia Challenge via miR-155 Suppression. (12th March 2022)

• Record Type: Journal Article
• Authors: Xue, Ming; Zhang, Xiwen; Chen, Jianxiao; Liu, Feng; Xu, Jingyuan; Xie, Jianfeng; Yang, Yi; Yu, Weiping; Qiu, Haibo
• Other Names: Najimi Mustapha Academic Editor.
• Is Part Of: Stem cells international. Volume 2022(2022)
• Publication Date: 2022-03-12
• Subjects: Stem Cells -- Periodicals; Stem Cells -- Therapeutic use -- Periodicals; Stem Cells -- Transplantation -- Periodicals; 616.0277405
• DOI: 10.1155/2022/5522828
• Languages: English
• ISSNs: 1687-966X

Abstract

Abstract : Background . Regulatory T cell (Treg)/T helper (Th) 17 skewing is important in the development of acute respiratory distress syndrome (ARDS). Immunomodulatory effects of mesenchymal stem cell- (MSC-) secreted transforming growth factor- (TGF-) β 1 on CD4 + T cells are environment-sensitive and lack discussion in hypoxic and inflammatory conditions. Methods . Mouse splenic CD4 + T cells were precoated with anti-CD3 (5 μ g/ml) and anti-CD28 (2 μ g/ml) overnight. RAW264.7 cells were added as antigen-presenting cells (APCs). T cells with and without RAW264.7 cells were treated with various LPS concentrations of 0, 10, 100, and 1000 ng/ml or/and at hypoxia condition of 5% O2 . Based on LPS (100 ng/ml) and hypoxia conditions (5% O2 ) as stimuli, MSCs were set as direct coculture or indirect coculture by transwell system. Anti-TGF- β 1 neutralization antibody was added to explore the role of TGF- β 1 among the soluble factors secreted by MSCs; miR-155 overexpression of CD4 + T cells was performed by transfection, and then, cells were added to the MSC-CD4 + T cell coculture system in hypoxic- and LPS-stimulated condition. After 48 hours, cells or supernatants were collected for detection of frequency of Treg and Th17 subsets, CD4 + T cell apoptosis and proliferation capacity assay by flow cytometry, secretion of INF- γ, IL-17A, IL-21, TGF- β 1, and IL-10 by ELISA, and levels of miR-155, Rorc, Foxp3, and Ptpn2 mRNA expression of CD4 + T cells by RT-PCR. Results . MSCs could restore skewed Treg/Th17 induced by LPS and hypoxia compared to groups without MSCs with increased secretion of TGF- β 1, IL-10, and IL-17A (P < 0.05 ) and attenuate the increased expression of miR-155 in CD4 + T cells via cell-to-cell contact mechanism while TGF- β 1 neutralization significantly inhibited the effects of MSCs restoring skewed Treg/Th17 and abolished its effect on miR-155 expression in CD4 + T cells. Conclusions . These findings suggested miR-155 suppression of CD4 + T cells mediated MSC-secreted TGF- β 1 modulating skewed Treg/Th17 induced by LPS-hypoxia challenge, providing evidence when proposing future T lymphocyte-targeted cell therapy in a specific condition.