Dataset Information

Autophagy enhances mesenchymal stem cell-mediated CD4⁺ T cell migration and differentiation through CXCL8 and TGF-?1.

ABSTRACT:

Background

Mesenchymal stem cells (MSCs) have been recognized as a promising tool for the treatment of various inflammatory disorders and autoimmune diseases. Stress conditions affect immune-mediated treatment and activate autophagy in MSCs. However, whether autophagy affects the MSC-mediated recruitment and differentiation of CD4⁺ T cells remains elusive.

Methods

MSCs were pretreated with 3-methyladenine (3-MA) and rapamycin to regulate autophagy, and then co-cultured with CD4⁺ T cells. CD4⁺ T cell migration and differentiation were detected by flow cytometry. Further, gene expression levels of well-known chemokines were analyzed by quantitative real-time PCR. Enzyme-linked immunosorbent assays and western blot analysis were performed to detect C-X-C motif chemokine ligand 8 (CXCL8) and transforming growth factor (TGF)-?1 protein levels. An exogenous antibody and short hairpin RNA were used to regulate CXCL8 and TGF-?1 levels, which enabled us to evaluate how autophagy affected MSC-mediated CD4⁺ T cell migration and differentiation.

Results

3-MA inhibited autophagy in MSCs, which was activated by rapamycin. Rapamycin increased the migration of CD4⁺ T cells, whereas 3-MA decreased their migration. Mechanistically, we found that autophagy strengthened CXCL8 secretion, and the addition of exogenous CXCL8 and an anti-CXCL8 antibody eliminated the difference of CD4⁺ T cell migration among groups. Further, the ratio of regulatory T (Treg) cells was increased in rapamycin-pretreated MSCs, but the ratio of T helper 1 (Th1) cells was decreased, while pretreatment of MSCs with 3-MA induced the opposite effect compared with the control group. TGF-?1 overexpression and knockdown using lentiviruses rectified the differences in the ratios of Treg and Th1 cells among the groups.

Conclusion

This study demonstrates that autophagy of mesenchymal stem cells mediates CD4⁺ T cell migration and differentiation through CXCL8 and TGF-?1, respectively. These results provide a potential new strategy for improving MSC-mediated therapy.

SUBMITTER: Cen S

PROVIDER: S-EPMC6708254 | biostudies-literature | 2019 Aug

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

Autophagy enhances mesenchymal stem cell-mediated CD4<sup>+</sup> T cell migration and differentiation through CXCL8 and TGF-β1.

Cen Shuizhong S Wang Peng P Xie Zhongyu Z Yang Rui R Li Jinteng J Liu Zhenhua Z Wang Shan S Wu Xiaohua X Liu Wenjie W Li Ming M Tang Su'an S Shen Huiyong H Wu Yanfeng Y

Stem cell research & therapy 20190823 1

<h4>Background</h4>Mesenchymal stem cells (MSCs) have been recognized as a promising tool for the treatment of various inflammatory disorders and autoimmune diseases. Stress conditions affect immune-mediated treatment and activate autophagy in MSCs. However, whether autophagy affects the MSC-mediated recruitment and differentiation of CD4<sup>+</sup> T cells remains elusive.<h4>Methods</h4>MSCs were pretreated with 3-methyladenine (3-MA) and rapamycin to regulate autophagy, and then co-cultured ...[more]

PMID: 31443687

Similar Datasets

Project description:BackgroundThe biological activity and regenerative medicine of bone marrow mesenchymal stem cells (BMSCs) have been focal topics in the broad fields of diabetic wound repair. However, the molecular mechanisms are still largely elusive for other cellular processes that are regulated during BMSC treatment. Our previous studies have shown that hypoxia is not only a typical pathological phenomenon of wounds but also exerts a vital regulatory effect on cellular bioactivity. In this study, the beneficial effects of hypoxic BMSCs on the cellular behaviors of epidermal cells and diabetic wound healing were investigated.MethodThe viability and secretion ability of hypoxic BMSCs were detected. The autophagy, proliferation and migration of HaCaT cells cultured with hypoxic BMSCs-derived conditioned medium were assessed by estimating the expression of autophagy-related proteins, MTS, EdU proliferation and scratch assays. And the role of the SMAD signaling pathway during hypoxic BMSC-evoked HaCaT cell autophagy was explored through a series of in vitro gain- and loss-of-function experiments. Finally, the therapeutic effects of hypoxic BMSCs were evaluated using full-thickness cutaneous diabetic wound model.ResultsFirst, we demonstrated that hypoxic conditions intensify HIF-1α-mediated TGF-β1 secretion by BMSCs. Then, the further data revealed that BMSC-derived TGF-β1 was responsible for the activation of epidermal cell autophagy, which contributed to the induction of epidermal cell proliferation and migration. Here, the SMAD signaling pathway was identified as downstream of BMSC-derived TGF-β1 to regulate HaCaT cell autophagy. Moreover, the administration of BMSCs to diabetic wounds increased epidermal autophagy and the rate of re-epithelialization, leading to accelerated healing, and these effects were significantly attenuated, accompanied by the downregulation of Smad2 phosphorylation levels due to TGF-β1 interference in BMSCs.ConclusionIn this report, we present evidence that uncovers a previously unidentified role of hypoxic BMSCs in regulating epidermal cell autophagy. The findings demonstrate that BMSC-based treatment by restoring epidermal cell autophagy could be an attractive therapeutic strategy for diabetic wounds and that the process is mediated by the HIF-1α/TGF-β1/SMAD pathway.

Dataset Information

Autophagy enhances mesenchymal stem cell-mediated CD4⁺ T cell migration and differentiation through CXCL8 and TGF-?1.

Background

Methods

Results

Conclusion

Publications

Autophagy enhances mesenchymal stem cell-mediated CD4<sup>+</sup> T cell migration and differentiation through CXCL8 and TGF-β1.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets

Dataset Information

Autophagy enhances mesenchymal stem cell-mediated CD4+ T cell migration and differentiation through CXCL8 and TGF-?1.

Background

Methods

Results

Conclusion

Publications

Autophagy enhances mesenchymal stem cell-mediated CD4<sup>+</sup> T cell migration and differentiation through CXCL8 and TGF-β1.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets

Autophagy enhances mesenchymal stem cell-mediated CD4⁺ T cell migration and differentiation through CXCL8 and TGF-?1.