The role of Toll-like receptor 1 in the immunoregulatory regulation of Th17 cells by mesenchymal stem cells / 中国组织工程研究

ABSTRACT

BACKGROUND:Our previous studies have shown that mesenchymal stem cells play an immunomodulatory role in Th17 cells, but the mechanism of action remains to be elucidated, therefore, to explore the role of Tol-like receptor 1 in which wil provide possible experimental basis for the future potential of celltherapy strategies. OBJECTIVE:To investigate the role of Tol-like receptor 1 in the immunoregulatory function of mesenchymal stem cells on Th17 cells. METHODS:Separation of adherent bone marrow-derived human embryonic sources of mesenchymal stem cells, immunomagnetic separation of normal CD4+T cells. CD4+T cells were cultured alone or in combination with mesenchymal stem cells co-cultured 4d. Real-time quantitative polymerase chain reaction assay interleukin-17, Tol-like receptor 1 expression levels related genes;number of Th17 cells by flow cytometry. Bone marrow mesenchymal stem cells from human embryos were separated using adherence method, and co-cultured with human CD4+T cells from healthy donors using immunomagnetic separation method for 4 days. The expression of interleukin-17 and Tol-like receptor 1 mRNA was detected by real-time PCR, and the number of Th17 cells was observed by flow cytometry. RESULTS AND CONCLUSION:Tol-like receptor 1 mRNA was expressed in both CD4+T cells and mesenchymal stem cells. Level of interleukin-17 mRNA was significantly higher in mesenchymal stem cells co-cultured with CD4+T cells than in CD4+T cells cultured alone (relative value 3.59±0.11 vs. 1.14±0.08, P<0.01). Consistent with the expression of interleukin-17 mRNA, increased level of Tol-like receptor 1 mRNA was detected in mesenchymal stem cells co-cultured with CD4+T cells compared with CD4+T cells cultured alone (relative value 6.07±1.79 vs. 1.53±0.63, P<0.01). Furthermore, Flow cytometry analysis showed that the percentage of Th17 cells in the mesenchymal stem cells co-cultured with CD4+T cells was significantly higher than that in CD4+T cells cultured alone, (4.53±1.27)%vs. (2.39±0.80)%(P<0.01). Tol-like receptor 1 might be involved in the immunoregulatory regulation of Th17 cells by mesenchymal stem cells.