Syk或ZAP-70信号途径诱导脐带血CD4^+T细胞极化  

Persistent and Selective Activation of Syk or ZAP-70 Precedes Th1 or Th2 On-Switch

作　　者：蒋艳萍[1] 杨大明[2] 吴静[1] 陈朗[1] 王俐[1] 谭锦泉[1] 

机构地区：[1]武汉大学医学院免疫系,湖北武汉430071 [2]湖北省监利县人民医院脑外科,湖北监利433300

出　　处：《武汉大学学报（医学版）》2008年第3期283-286,297,I0001,共6页Medical Journal of Wuhan University

基　　金：国家自然科学基金资助项目(编号:30572119)

摘　　要：目的:探讨IL-2或IL-4和基质细胞源因子(SDF)-1α协同刺激下,脐带血CD4+T细胞极化的作用及其与非受体型蛋白质酪氨酸激酶的关系。方法:流式细胞术、实时定量RT-PCR检测IL-2或IL-4和SDF-1α协同刺激下,CD4+T细胞中胞内Th1型和Th2型细胞因子的表达;免疫复合物激酶分析法和免疫印迹法检测Th细胞极化过程中非受体型蛋白酪氨酸激酶Syk和ZAP-70的磷酸化;反义肽核酸(PNA)技术探讨Syk和ZAP-70在协同刺激Th细胞极化过程中的作用。结果:L-2或IL-4和SDF-1α协同作用可通过Syk或ZAP-70磷酸化诱导CD4+T细胞极化;Syk或ZAP-70 PNA可明显阻断Syk或ZAP-70磷酸化以及CD4+T细胞极化。结论:Syk和ZAP-70磷酸化对于IL-2或IL-4和SDF-1α协同诱导CD4+T细胞极化十分重要。Objective： To address which signaling pathways might be involved in the on-switch of cord blood （CB） CD4＋Th cells by IL-2 or IL-4 and SDF-1α. Methods. Flow cytometry and real time quantitative RT-PCR were used to detect the intracellular Thl and Th2 cytokine and cytokine mRNA. The immune complex kinase assay （KA） and immunoblotting （IB） were utilized to de- termine the activation of Syk and ZAP-70 in the CB CD4^＋ T cells which were either freshly isolated or co-stimulated with IL-2 or IL-4 and SDF-1α. The immune complex kinase assay and immu-noblotting were also exploited to analyze the blocking effects of peptide nucleic acid （PNA） Syk or ZAP-70 antisenses on the activation of Syk or ZAP-70 in CB CD4^＋ T cells. Results： IL-2 or IL-4 and SDF-1α can induce CB CD4^＋T cells to switch to Th1 or Th2 through phosphorylating Syk or ZAP-70. Syk or ZAP-70 PNA can significantly inhibit kinase activity and expression of cyto-kines. Conclusion： Persistent and selective activation of Syk or ZAP-70 is essential for Th polari-zation.

关 键 词：白细胞介素-2 白细胞介素-4 基质细胞源因子-1α 非受体型蛋白酪氨酸激酶 反义肽核酸 

分 类 号：R392.1[医药卫生—免疫学]