Objective To evaluate the effect of suppressive oligodeoxynucleotides (Sup ODN) on expression of T-bet, pSTAT4 and pSTAT6 of CD4 + T spleen lymphocytes in vitro. Methods The spleen lymphocytes of BALB/c mice were isolated and then CD4 + T cells were purified with immunomagnetic microbeads( positive selection), the purification was analysed by flow cytometry. The co-culture of CD4 + cells, anti-CD3e, anti-CD28, IL-12 and Sup ODN or control oligodeoxynucleotides(Con ODN) was performed for 72 hours, the nucleoprotein was extracted with regular procedure. Expression of pSTAT4(phosphorylated STAT4) ,pSTAT6,and T-bet in nucleoprotein were detected by Western-blot. Results Compared with Con ODN and PBS treated group, expression of T-bet and pSTAT4 in Sup ODN treated group decreased significantly (P<0. 05), while expression of pSTAT6 in Sup ODN treated group increased significantly (P<0. 05). Conclusion Sup ODN may inhibit Thl differentiation of mice CD4 + T cell by down-regulating expression of pSTAT4 and T-bet, and promote Th2 differentiation by up-regulating expression of pSTAT6.%目的 探讨体外实验中,抑制性寡脱氧核苷酸(Sup ODN)对小鼠脾脏CD4+T细胞转录因子T细胞表达的T盒(T-bet)、信号转导子和转录激动子(STAT)4及STAT6表达的影响.方法 分离小鼠脾脏淋巴细胞,免疫磁珠阳选法纯化CD4+T细胞,流式细胞仪分析分选纯度,在抗-CD3ε抗体、抗-CD28抗体、白细胞介素(IL)-12作用下,加入Sup ODN或对照寡脱氧核苷酸(Con ODN)培养72 h后,常规提取核蛋白.Western-blot测定核蛋白中磷酸化的STAT(pSTAT)4、pSTAT6和T-bet蛋白表达.结果 与Con ODN组和PBS组比较,Sup ODN 组中T-bet和pSTAT4的表达水平明显降低(P＜0.05),而pSTAT6表达水平明显升高(P＜0.05).结论 Sup ODN可能通过下调pSTAT4和T-bet的表达,体外抑制小鼠CD4+T细胞向Th1细胞的功能性分化,上调pSTAT6的表达,促进Th2细胞的分化.
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