TNF-α-induced apoptosis in neonatal lymphocytes: TNFRp55 expression and downstream pathways of apoptosis
TNF-α-induced apoptosis in neonatal lymphocytes: TNFRp55 expression and downstream pathways of apoptosis
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DOI：
10.1038/sj.gene.6363674
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发表时间：
2000-04-01
影响因子：
5
通讯作者：
Gupta, S
中科院分区：
文献类型：
--
作者：
Aggarwal, S;Gollapudi, S;Gupta, S
以前，我们已经表明减少Fas介导的细胞凋亡在脐带血淋巴细胞亚群。在这项研究中，我们比较了肿瘤坏死因子（TNF）-α诱导的T淋巴细胞及其亚群之间的脐带血和外周血健康的年轻人对照组的凋亡。流式细胞术和定量RT-PCR检测TNF受体I（TNFR-I）的表达。Western blotting分析接头分子TNF受体相关死亡结构域（TRADD）、Fas相关死亡结构域（FADD）、TNF相关因子2（TRAF-2）和caspase 3的表达。比色法测定caspase 3和caspase 8活性。用末端脱氧核苷酸转移酶介导的dUTP缺口末端标记法（TUNEL）检测CD 4（+）和CD 8（+）T细胞对TNF-α诱导的凋亡的敏感性。与外周血T细胞亚群相比，脐带血CD 4（+）和CD 8（+）T细胞亚群对TNF-α诱导的细胞凋亡的敏感性降低，这与caspase 8和caspase 3的活化降低有关。此外，与外周血淋巴细胞相比，脐带血淋巴细胞中TNFR-I、TRADD和caspase 3的表达降低。这些意见的意义进行了讨论。
Previously we have shown decreased Fas-mediated apoptosis in cord blood lymphocyte subsets. In this study, we compared tumor necrosis factor (TNF)-alpha-induced apoptosis in T lymphocytes and their subsets between cord blood and peripheral blood from healthy young controls. The expression of TNF receptor I (TNFR-I) was assessed by flow cytometry and quantitative RT-PCR. The expression of adapter molecules TNF receptor-associated death domain (TRADD), Fas-associated death domain (FADD) and TNF-associated factor-2 (TRAF-2) and caspase 3 was analyzed by Western blotting. The activity of caspase 3 and caspase 8 was measured by colorimetric assay. The susceptibility of CD4(+) and CD8(+) T cells to TNF-alpha-induced apoptosis was measured by terminal deoxytidyl transferase-mediated dUTP nick end labelling (TUNEL) assay. Both CD4(+) and CD8(+) T cell subsets from cord blood demonstrated decreased susceptibility to TNF-alpha-induced apoptosis that was associated with decreased activation of both caspase 8 and caspase 3 as compared to T cell subsets in peripheral blood. Furthermore, expression of TNFR-I, TRADD and caspase 3 was decreased in cord blood lymphocytes as compared to peripheral blood lymphocytes. The significance of these observations is discussed.