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信号淋巴细胞激活分子(SLAM)在人Th1和Th2细胞中呈差异表达。

Signaling lymphocytic activation molecule (SLAM) is differentially expressed in human Th1 and Th2 cells.

作者信息

Hamalainen H, Meissner S, Lahesmaa R

机构信息

Roche Bioscience, 3401 Hillview Avenue, Palo Alto, CA 94-301, USA.

出版信息

J Immunol Methods. 2000 Aug 28;242(1-2):9-19. doi: 10.1016/s0022-1759(00)00200-3.

Abstract

We have used a real-time quantitative RT-PCR technique (TaqMan, PE Biosystems) to identify genes that are differentially expressed by human polarised CD4(+) T cell subsets (Th1 or Th2). The goal was to test the feasibility of the detection method in profiling the expression of a set of marker genes important for Th1 and Th2 differentiation. We demonstrate that in polarised human Th1 cells signaling lymphocytic activation molecule (SLAM), a member of the immunoglobulin superfamily, is expressed at 7-25-fold higher levels than in Th2 cells. Along with SLAM, expression of the IL-12 receptor chain beta 2 (IL-12R beta 2) and the IFN-gamma receptor chain beta (IFN-gamma R beta) proved to be useful molecular markers indicating the state of T cell polarisation, as previously reported. Treatment with IL-12 increased SLAM mRNA expression in T cells by 3-4-fold, whereas a number of other cytokines including PDGF-BB, IFN-alpha A, IFN-alpha A/D, IFN-beta, IFN-gamma or IL-9 had no effect. Stimulating T cells by co-ligating CD3 and CD28 increased SLAM protein surface expression in both Th1 and Th2 cells. In conclusion, real-time RT-PCR detection was found to be an accurate, sensitive and highly reproducible method for fast profiling of mRNA expression in Th1 and Th2 cell subsets.

摘要

我们使用了实时定量逆转录聚合酶链反应技术(TaqMan,PE Biosystems公司)来鉴定在人极化CD4(+) T细胞亚群(Th1或Th2)中差异表达的基因。目的是测试该检测方法在分析一组对Th1和Th2分化重要的标记基因表达方面的可行性。我们证明,在极化的人Th1细胞中,免疫球蛋白超家族成员信号淋巴细胞激活分子(SLAM)的表达水平比Th2细胞高7至25倍。与SLAM一样,如先前报道的,IL-12受体链β2(IL-12Rβ2)和IFN-γ受体链β(IFN-γRβ)的表达被证明是指示T细胞极化状态的有用分子标记。用IL-12处理可使T细胞中SLAM mRNA表达增加3至4倍,而包括血小板衍生生长因子-BB、IFN-αA、IFN-αA/D、IFN-β、IFN-γ或IL-9在内的许多其他细胞因子则无作用。通过共连接CD3和CD28刺激T细胞可增加Th1和Th2细胞中SLAM蛋白的表面表达。总之,实时逆转录聚合酶链反应检测被发现是一种准确、灵敏且高度可重复的方法,用于快速分析Th1和Th2细胞亚群中的mRNA表达。

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