The Pattern And Regulation Mechanism Of L-Selectin Expression Of Activated Human γδT Cells Induced By Stimulating With Mycobacterium Tuberculosis Antigen

Objective (1) To explore dynamic expression of L-selectin(CD62L) in human peripheral γδT cells activated with Mycobacterium tuberculosis antigen(Mtb-Ag) and cultured for proliferation. (2) To investigate effects of different cytokine on the expression of L-selectin of human γδT cells. (3) To investigate the roles of PKC, PI3K,and MAPK pathways in the regulation of L-selectin expression of activated humanγδT cells induced by restimualting with Mtb-Ag.Methods (1) The peripheral blood mononuclear lymphocytes (PBMC) of healthy donors were stimulated with Mtb-Ag and CD3mAb, cultured in IL-2 containing media, to generate Mtb-Ag activated T cells (MtbAT) (γδT cells were predominant) and CD3mAb activated T cells (CD3AT) (αβT cells were predominant) respectively. (2) After stained with multicolor fluorescence mAbs, the percentage of L-selectin among isolation fresh PBMC, MtbAT or CD3AT cultured for 3 to 21 days were measured by flow cytometry (FCM). (3) Human γδT cells activated by primary and secondary stimulation with Mtb-Ag were cultured for 0 days, 3 days, 5 days, and 7days, without or with cytokines IFN-γ, anti-IFN-γ, IL-12 or anti-IL-12,the expression of L-selectin on γδT cells were detected by FCM. (4) MtbAT which were cultured for 6 days to 8 days were restimulated with PMA, Ionomycin(IMN), PMA+IMN respectively for 3 hours, 6 hours, 12 hours, 24 hours, and the expression of L-selectin on...