Distribulion of interleukin-22 producing cells in lymphocyte subpopulations from peripheral blood

Objective:To establish a method for detection of interleukin-22 producing cells in lymphocyte subpopulations from peripheral blood by flow cytometry.Methods:Peripheral blood samples were mixed with same volume of RPMI 1640 medium and stimulated with PMA and ionomycin for 6 hours in the presence of monensin at last 4 hours at 37 ℃,5% CO2.The stimulated cells were stained for surface molecules or intracellular cytokines with fluorochrome-conjugated mAbs.The proportions of IL-22 producing cells and IL-17 producing cells among different lymphocyte subsets were detected by flow cytometry.Results:The proportions of IL-22 producing cells among CD3+T cells,CD4+T cells,CD8+T cells,NK cells and B lymphocytes were 1.83%,2.12%,0.67%,1.96% and 1.09%,respectively.The amount of IL-22 producing cells among CD4+T cells was significantly higher than that among CD8+T cells(P0.01).In addition,IL-22 and IL-17 co-expressing cells were also detected in CD3+T cells,CD4+T cells,and CD8+T cells.Conclusions:By using flow cytometry technique,the IL-22 producing cells exist among all CD4+T cells,CD8+T cells,NK cells and B lymphocytes.The proportion of IL-22 producing cells in CD4+T cells is higher than that in CD8+T cells.