PDIA3对人Jurkat T细胞TCR信号通路的调控

为了研究蛋白质二硫键异构酶A3前体(protein disulfide isomerase A3 precursor, PDIA3)在T细胞受体(T cell receptor, TCR)信号通路中的具体功能,利用电穿孔法将T 细胞内的PDIA3蛋白水平敲低,通过Western blotting检测ζ-链相关蛋白70(zeta-chain associated protein 70, ZAP70)的磷酸化修饰水平, 酶联免疫吸附测定(enzyme linked immunosorbent assay, ELISA)实验检测细胞因子IL-2的分泌情况,流式细胞术分析分化抗原簇69(cluster of differentiation 69, CD69)表达水平及荧光素酶报告基因检测NF-κB信号通路.结果显示:T细胞中PDIA3蛋白下调后导致ZAP70蛋白的磷酸化水平、CD69表达和IL-2的分泌都明显降低,并且影响了NF-κB信号通路,表明PDIA3蛋白对T细胞的活化有促进作用,参与T细胞TCR信号通路的调控,为进一步深入研究PDIA3与TCR下游一些功能蛋白的相互作用打下了基础.

PDIA3 regulates TCR signaling in human Jurkat T cells

In order to study the specific function of protein disulfide isomerase A3 precursor(PDIA3)in T cell receptor(TCR)signaling pathway, the PDIA3 protein level in T cells was knocked down by electroporation, the phosphorylation level of zeta-chain associated protein 70( ZAP70)protein was detected by enzyme linked immunosorbent assay(ELISA), and the expression level of cluster of differentiation 69(CD69)were analyzed by flow cytometry. The luciferase reporter gene detected NF-κB signaling pathway. The results showed that down-regulation of PDIA3 protein in T cells resulted in a significant decrease in ZAP70 protein phosphorylation, CD69 expression, IL-2 secretion, and affected NF-κB signaling pathway, indicating PDIA3 protein promotes the activation of T cells and participate in the regulation of T cell TCR signaling pathway. This work provides an important research basis for studying the interaction between PDIA3 and some functional proteins downstream of TCR in the future.