Neutrophil-derived TNF-related apoptosis-inducing ligand (TRAIL): A novel mechanism of antitumor effect by neutrophils
Neutrophil-derived TNF-related apoptosis-inducing ligand (TRAIL): A novel mechanism of antitumor effect by neutrophils
复制标题
DOI：
10.1158/0008-5472.can-03-1808
复制
发表时间：
2004-02-01
期刊：
影响因子：
11.2
通讯作者：
Hara, T
中科院分区：
文献类型：
--
作者：
Koga, Y;Matsuzaki, A;Hara, T
为了检测中性粒细胞中独特表达的新基因并阐明其功能，我们利用含有240个细胞因子基因的cDNA芯片比较了中性粒细胞和健康人外周血单个核细胞（PBMCs）之间的基因表达模式。包括肿瘤坏死因子相关凋亡诱导配体（TPAIL）在内的26个基因在中性粒细胞中的表达水平比植物血凝素刺激的PBMCs高10倍。real-time逆转录- pcr和ELISA分别测定TRAIL mRNA和蛋白的表达量。TRAIL在静息中性粒细胞中以mRNA和蛋白水平表达，ifn - γ刺激后其表达增强。中性粒细胞在细胞表面表达TRAIL并将其释放到培养基中。使用fitc偶联膜联蛋白v，流式细胞术检测中性粒细胞来源的TRAIL对Jurkat细胞的细胞毒性。当Jurkat细胞与中性粒细胞在ifn - γ存在下培养时，发生凋亡的Jurkat细胞数量增加，这种增加取决于效应：靶比。在培养基中加入抗trail抗体可部分抑制这种细胞毒性。中性粒细胞可能通过TRAIL发挥自身的抗肿瘤作用。微阵列分析被发现是一种有用的工具，用于检测被认为在中性粒细胞功能中起未知作用的新基因。
To detect the novel genes expressed uniquely in neutrophils and elucidate their function, the gene expression pattern was compared by using cDNA microarray containing 240 cytokine genes between the neutrophils and peripheral blood mononuclear cells (PBMCs) obtained from healthy human donors. Twenty-six genes, including tumor necrosis factor-related apoptosis-inducing ligand (TPAIL), were expressed in neutrophils at a level >10 times higher than that seen in phytohemagglutinin-stimulated PBMCs. The amounts of mRNA and protein of TRAIL were quantified by real-time reverse transcription-PCR and ELISA, respectively. TRAIL was expressed in resting neutrophils at the mRNA and protein levels, and its expression was enhanced after stimulation with IFN-gamma. Neutrophils expressed TRAIL on the cell surface and released it into the culture media. The cytotoxicity of neutrophil-derived TRAIL against Jurkat cells was determined by flow cytometry using FITC-conjugated annexin V. When Jurkat cells were cultured with neutrophils in the presence of IFN-gamma, the number of Jurkat cells undergoing apoptosis increased, and such increase depended on the effector:target ratio. This cytotoxicity was suppressed partially by adding anti-TRAIL antibody to the media. Neutrophils may exert their own antitumor effect by TRAIL. A microarray analysis was found to be a useful tool for detecting novel genes that are suggested to play unknown roles in the neutrophil function.