Chinese Journal of Tissue Engineering Research ›› 2021, Vol. 25 ›› Issue (31): 4928-4938.doi: 10.12307/2021.132

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Immunomodulatory effects of umbilical cord-mesenchymal stem cells modified by miR-1-5p on T lymphocyte subsets in systemic lupus erythematosus

Hu Mingzhi1, Zhang Jingying2, Yang Guoan3, Pang Chunyan3, Zhang Wei3, Wang Yongfu3, Sun Xiaolin3   

  1. 1Department of Rheumatology, First Affiliated Hospital of Baotou Medical College, Institute of Rheumatology of Baotou Medical College, Baotou 014010, Inner Mongolia Autonomous Region, China; 2Department of Critical Care Medicine, First Affiliated Hospital of Baotou Medical College, Baotou 014010, Inner Mongolia Autonomous Region, China; 3Key Laboratory of Autoimmunology, Baotou 014010, Inner Mongolia Autonomous Region, China 

  • Received:2020-08-13 Revised:2020-08-14 Accepted:2020-09-26 Online:2021-11-08 Published:2021-04-25
  • Contact: Sun Xiaolin, MD, Laboratorian-in-charge, Key Laboratory of Autoimmunology, Baotou 014010, Inner Mongolia Autonomous Region, China
  • About author:Hu Mingzhi, Master, Department of Rheumatology, First Affiliated Hospital of Baotou Medical College, Institute of Rheumatology of Baotou Medical College, Baotou 014010, Inner Mongolia Autonomous Region, China
  • Supported by:
    National Natural Science Foundation of China, No. 81860294 (to SXL), No. 81860295 (to ZW); Natural Science Foundation of Inner Mongolia Autonomous Region, No. 2019MS08055 (to SXL); Science and Technology Project of Inner Mongolia Autonomous Region, No. 201802089 (to SXL), No. 2019GG052 (to WYF)

Abstract: BACKGROUND: In recent years, there were lots of researches on the treatment of diseases by gene modified mesenchymal stem cells, including the treatment of autoimmune diseases. Various miRNAs have been found to be abnormally expressed in autoimmune diseases, among which miR-1 is low-expressed in polymyositis, and miR-1-3p is one of the miRNAs related to systemic lupus eythematosus susceptibility genes.  
OBJECTIVE: To investigate the immunoregulatory effect of miR-1-5p modified umbilical cord-mesenchymal stem cells on T lymphocyte subsets in systemic lupus erythematosus.
METHODS:  The expression level of miR-1-5p in the two groups was detected by real-time fluorescence quantitative PCR in eight patients aged 30-60 years with active systemic lupus erythematosus and eight healthy subjects. Umbilical cord-mesenchymal stem cells were isolated and cultured from umbilical cord of newborns delivered at term or cesarean section under aseptic conditions. Using the liposome method, the umbilical cord-mesenchymal stem cells were transfected with Red Fluorescent Control with a concentration gradient of 5, 7, 10, and 15 nmol/L for 24, 48, and 72 hours. Transfection efficiency of miR-1-5p was determined by flow cytometry at different concentration gradients and time gradients. Peripheral blood mononuclear cells in systemic lupus erythematosus patients were isolated by density gradient centrifugation and co-cultured with umbilical cord-mesenchymal stem cells which were transfected with miR-1-5p, and divided into four groups: umbilical cord-mesenchymal stem cells transfected with miR-1 and co-cultured with peripheral blood mononuclear cells of systemic lupus erythematosus patients, umbilical cord-mesenchymal stem cells transfected with miRNA negative control and co-cultured with peripheral blood mononuclear cells of systemic lupus erythematosus patients, umbilical cord-mesenchymal stem cells co-cultured with peripheral blood mononuclear cells of systemic lupus erythematosus patients, and peripheral blood mononuclear cells of systemic lupus erythematosus patients without treatment group. The co-culture was conducted for 48 hours. The relative expression levels of IL-17A, Foxp3, IFN-γ and IL-4 genes in each group were detected by real-time fluorescence quantitative PCR. Flow cytometry was used to detect the differentiation of Th17, Treg, Th1 and Th2 cells in peripheral blood mononuclear cells of each group. 
RESULTS AND CONCLUSION: (1) miR-1-5p was significantly down-regulated in peripheral blood mononuclear cells of systemic lupus erythematosus patients compared with healthy controls (P < 0.05). (2) The transfection efficiency was highest when the concentration of miR-1-5p was 10 nmol/L and the transfection time was 48 hours. (3) Compared with the peripheral blood mononuclear cells of systemic lupus erythematosus patients without treatment group, the expression of Foxp3 in umbilical cord-mesenchymal stem cells transfected with miR-1-5p and co-cultured with peripheral blood mononuclear cells of systemic lupus erythematosus patients group was increased; the proportion of Treg cells was significantly increased; and the differences were statistically significant (P < 0.05). Compared with the peripheral blood mononuclear cells of systemic lupus erythematosus patients without treatment group, Th17/Treg ratio decreased in the other three co-culture groups, and the difference was statistically significant (P < 0.01). Th17/Treg cell ratio detected by flow cytometry in each group was not significantly different (P > 0.05). Compared with the peripheral blood mononuclear cells of systemic lupus erythematosus patients without treatment group, the Th1/Th2 ratio in the umbilical cord-mesenchymal stem cells transfected with miR-1-5p and co-cultured with PBMC of systemic lupus erythematosus patients group decreased, and the difference was statistically significant (P < 0.05). However, the Th1/Th2 cell ratio of each group detected by flow cytometry was not significantly different (P > 0.05). (4) The results showed that the umbilical cord-mesenchymal stem cells modified by miR-1-5p had an immunoregulatory effect on Treg cells of systemic lupus erythematosus, and overexpression of miR-1-5p could promote the differentiation and proliferation of Treg cells of systemic lupus erythematosus patients. miR-1-5p regulates the immune imbalance of Th17/Treg and Th1/Th2 by up-regulating the expression of Treg cells. 


Key words: stem cells, umbilical cord-mesenchymal stem cells, miR-1, systemic lupus erythematosus, T lymphocytes, Treg cells, Th1, Th2, immunoregulation

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