蛋白激酶Cα对T细胞生物学功能影响的研究

Roles of PKCα on the biological functions of T cells

摘要:

目的探讨蛋白激酶Cα(PKCα)对T细胞增殖、凋亡、分化、细胞因子的生成、诱导性调节性T细胞(iTreg)生成的影响。方法分离野生型(PKCα+/+组)或PKCα基因敲除(PKCα-/-组)小鼠T细胞,体外培养,在T细胞表面受体(TCR)信号刺激下,利用3H胸腺嘧啶掺入法、CSFE/Annexin V染色结合流式细胞技术检测T细胞增殖及凋亡情况;收集细胞培养上清,用ELISA方法检测细胞因子IL-2、IL-4、IFN-γ和IL-17的生成。分离PKCα+/+组或PKCα-/-组小鼠CD4+T细胞,在TCR信号刺激下,按Th17细胞、iTreg分化条件进行体外诱导,用流式细胞技术检测细胞分化结果。结果与PKCα+/+组相比,PKCα缺失时,在TCR信号刺激下,T细胞的增殖降低,IL-2生成增多,IL-4和IL-17生成减少,Th17细胞分化减少,iTreg生成增加(均P<0.05)。结论 PKCα具有促炎作用。

Abstract:

Objective To study the roles of PKCα on the proliferation, apoptosis, differentiation, cytokine production and inducible regulatory T cell (iTreg) induction of T cells. Methods T cells from WT (PKCα+/+) or PKCα knockout (PKCα-/-) mice were isolated and cultured in vitro. T cell proliferation and apoptosis were determined using 3H thymidine incorporation and CSFE/Annexin V staining. Cytokines production (IL-2, IL-4, IFN-γ and IL-17) was detected using ELISA. CD4+T cells were isolated and cultured in vitro via Th17 or iTreg biased condition. Flow cytometry was used to detect the cell differentiation. Results The production of IL-2 upon TCR stimulation increased, while the contents of IL-4 and IL-17 decreased in the PKCα-/- group compared with the PKCα+/+ group. The differentiation rate of Th17 cells decreased, while the iTreg production increased in the PKCα-/- group compared with the PKCα+/+ group. Conclusions PKC-α is proinflammatory.