In vitro co-induction of Th17 cells by using MCMV and PMA/Ionomycin

To investigate the co-induction of Th17 cells in vitro by using MCMV and PMA/Ionomycin,the splenocytes of BALB/c mice were stimulated with MCMV or PMA/Ionomycin or both of MCMV and PMA/Ionomycin.The proportion of CD4+IL-17+ cells in splenocytes was analyzed by flow cytometry and mean fluorescent intensity was also investigated.Meanwhile,the level of IL-17 in culture supernatant was detected by ELISA.Our results showed that usage of MCMV alone or PMA/Ionomycin to stimulate spenocytes could not induce Th17 cell differentiation efficiently and very lower proportion of CD4+IL-17+ cells was found.However,the use of MCMV and PMA/Ionomycin could obviously induce Th17 cell differentiation and the proportion of CD4+IL-17+ cells reached up to 2.6%,while the use of MCMV and PMA/Ionomycin could also significantly increase the mean fluorescent intensity of CD4+IL-17+ cells.Furthermore,the use of both MCMV and PMA/Ionomycin could remarkably increase the level of IL-17 in culture supernatants.Our results can establish the experimental foundation for the further study on the relationship between Th17 cells and the CMV infection.