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目的:研究金黄色葡萄球菌肠毒素A(SEA)对K562细胞体外诱导脐血T细胞活化TCRζ链基因表达情况。方法:常规分离4例脐带血单核细胞,分别与抗CD3单克隆抗体、单纯K562细胞、SEA以及SEA联合K562细胞共培养,诱导T细胞活化增殖,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cD-NA,采用SYBR GreenⅠ荧光定量PCR和相对定量检测TCRζ链在不同组别T淋巴细胞中的表达情况,以β2微球蛋白基因(β2M)作为内参,根据相对定量公式:2-ΔΔCt计算TCRζ链表达差异倍数。结果:抗CD3单抗组、K562细胞组、SEA组、SEA联合K562细胞组诱导培养T细胞中TCRζ链表达差异倍数分别是(4.52±0.96)、(1.65±0.26)、(1.43±0.44)、(3.41±0.30),表明各组均有活化T细胞的作用,但各组TCRζ链表达水平有差异,其中SEA联合k562细胞组的T细胞ζ链基因表达均明显高于单纯k562组及单纯SEA组(P<0.01)。结论:超抗原SEA有助于增强K562细胞体外诱导T细胞活化作用。
Objective: To study the effect of Staphylococcus aureus enterotoxin A (SEA) on the expression of activated TCRζ chain gene in cord blood T cells induced by K562 cells in vitro. Methods: Four cases of umbilical cord blood mononuclear cells were isolated routinely and were respectively cultured with anti-CD3 monoclonal antibody, K562 cells alone, SEA and SEA combined with K562 cells to induce T cell activation and proliferation, and a blank control group was established. After 48 h of culture, mRNA of each group was collected and cD-NA was synthesized. The expression of TCRζ chain in different groups of T lymphocytes was detected by SYBR Green Ⅰ fluorescence quantitative PCR and relative quantification. Β2 microglobulin gene (β2M) As an internal reference, according to the relative quantification formula: 2-ΔΔCt TCRζ chain expression difference multiples. RESULTS: The differences in TCRζ chain expression in T cells induced by anti-CD3 McAb, K562 cells, SEA group and SEA combined with K562 cells were (4.52 ± 0.96), (1.65 ± 0.26) and (1.43 ± 0.44) (3.41 ± 0.30), indicating that all groups have the role of activating T cells, but the expression of TCR ζ chain in each group is different. SEA combined with k562 cells T-cell ζ chain gene expression were significantly higher than the k562 group and SEA alone Group (P <0.01). Conclusion: Superantigen SEA can enhance the activation of T cell induced by K562 cells in vitro.