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目的:探讨金黄色葡萄球菌肠毒素A(SEA)对K562细胞诱导正常人脐带血单个核细胞(MNCs)中CD3ε链基因表达的影响。方法:常规分离4例脐带血单个核细胞,分别与抗CD3单克隆抗体(mAb)、单纯K562细胞、单纯SEA以及SEA联合K562细胞共培养,诱导MNCs活化增殖,并设空白对照组。刺激培养48 h后,收集各组细胞提取mRNA并合成cDNA,以β2微球蛋白基因作为内参照,采用实时荧光定量PCR检测在各组MNCs中CD3ε链基因的表达,并根据公式2-△△C t对CD3ε链表达的差异倍数进行相对定量分析。结果:K562细胞组MNCs中CD3ε链基因表达的水平略有降低,抗CD3 mAb组、SEA组、SEA联合K562细胞组的诱导活化的MNCs中CD3ε链基因的表达均有增强,而SEA联合K562细胞组MNCs中CD3ε链基因的表达明显高于单纯SEA组(P<0.01)。结论:超抗原SEA可以增加K562细胞在体外诱导脐带血MNCs中CD3ε链基因的表达。
Objective: To investigate the effect of Staphylococcus aureus enterotoxin A (SEA) on the gene expression of CD3ε in normal human cord blood mononuclear cells (MNCs) induced by K562 cells. Methods: Umbilical cord blood mononuclear cells were isolated routinely. MNCs were induced to proliferate by co-culture with anti-CD3 monoclonal antibody (mAb), K562 cells alone, SEA alone and SEA combined with K562 cells. After stimulated for 48 hours, mRNA was collected from each group and cDNA was synthesized. Β2 microglobulin gene was used as internal reference, and the expression of CD3ε chain in MNCs of each group was detected by real-time fluorescence quantitative PCR. According to the formula 2- △△ C t relative expression of CD3 epsilon multiple fold relative quantitative analysis. Results: The expression of CD3ε chain gene in MNCs of K562 cells decreased slightly. The expression of CD3ε chain gene in MNCs induced by anti-CD3 mAb, SEA and SEA combined with K562 cells increased. However, SEA combined with K562 cells The expression of CD3ε chain gene in MNCs was significantly higher than that in SEA alone group (P <0.01). Conclusion: Superantigen SEA can increase the expression of CD3ε chain gene in cord blood MNCs induced by K562 cells in vitro.