抑制性寡脱氧核苷酸对小鼠CD4+T细胞Tbet、STAT4及STAT6表达的影响
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范学工

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R392.11

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国家自然科学基金(30671845)教育部高校博士点基金新教师资助项目(20070533009)
湖南省卫生厅科研基金资助项目 (B2005030)中南大学大学生创新性实验计划项目(071053365)


Effect of suppressive oligonucleotides on mice CD4+T cell transcription factor and its significance
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    摘要:

    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。

    Abstract:

    ObjectiveTo evaluate the effect of suppressive oligodeoxynucleotides (Sup ODN) on expression of Tbet, pSTAT4 and pSTAT6 of CD4+T spleen lymphocytes in vitro.MethodsThe 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CD3ε, 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.ResultsCompared 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).ConclusionSup ODN may inhibit Th1 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.

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向瑛,范学工,等.抑制性寡脱氧核苷酸对小鼠CD4+T细胞Tbet、STAT4及STAT6表达的影响[J]. 中国感染控制杂志,2011,10(5):331-335.
XIANG Ying, FAN Xuegong, et al. Effect of suppressive oligonucleotides on mice CD4+T cell transcription factor and its significance[J]. Chin J Infect Control, 2011,10(5):331-335.

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  • 收稿日期:2011-07-12
  • 最后修改日期:2011-07-31
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  • 在线发布日期: 2011-09-30
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