Adipose-derived stem cells mediate immunosuppression of Th17 in multiple sclerosis |
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| Authors: | LIN Xiao-bin CHEN Ying YANY De-hao XIE Yong-lin BI Yong KE Jian-ming CHEN Zhi-bo SU Zhong-qian LI Xiang ZHANG Xu |
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| Institution: | 1. Department of Ultrasound, The First Affiliated Hospital, Wenzhou Medical University, Wenzhou 325000, China;
2. Department of Neurology, The First Affiliated Hospital, Wenzhou Medical University, Wenzhou 325000, China;
3. Department of Rehabilitation, Zhejiang Provincial Hospital of Chinese Armed Police Force, Hangzhou 310000, China |
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| Abstract: | AIM: To investigate how human adipose-derived stem cells (hASCs) regulates the differentiation of Th17 cells in multiple sclerosis. METHODS: hASCs were isolated from the adipose tissues. Magnetic-activated cell sorting (MACS) kit was used to isolate CD4+ T cells from peripheral blood mononuclear cells (PBMCs) which were isolated by density gradient centrifugation. The percentage of CD4+ T cells was detected by flow cytometry. The activated CD4+ T cells were co-cultured with hASCs for about 4 d at different ratios of hASCs to CD4+ T cells (1:4 and 1:10) in a Th17 polarised condition. Another group adding anti-leukemia inhibitory factor (LIF) antibody was set up. Th17 cell proportion of the CD4+ T cells was determined by flow cytometry. The level of LIF in the supernatant of co-cultured system was measured by ELISA. The mRNA expression of retinoid-related orphan receptor γt (RORγt), interleukin-6 receptor (IL-6R), interleukin-23 receptor (IL-23R), LIF and leukemia inhibitory factor receptor (LIFR) was detected by real-time PCR. RESULTS: The result of flow cytometry suggested there were mainly hASCs, and the percentage of CD4+ T cells in the PBMCs were above 90% after MACS. The Th17 cell proportion decreased in 1:4 and 1:10 co-cultured groups in a dose-dependent manner. The mRNA expression of IL-6R, IL-23R and RORγt was downregulated and the expression of LIFR and LIF was up-regulated. When the anti-LIF was added into the co-cultured system, the ratio of Th17 cells increased and reached to the control level. The protein level of LIF obviously increased after co-cultured. After anti-LIF added, the mRNA expression of RORγt and IL-6R was up-regulated. CONCLUSION: hASCs inhibits the differentiation of Th17 cells from multiple sclerosis patients through the competitive inhibition of LIF/IL-6 by secreting LIF. |
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| Keywords: | Multiple sclerosis Adipose-derived stem cells Th17 cells Leukemia inhibitory factor Interleukin-6 |
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