TNF‐related apoptosis‐inducing ligand (TRAIL) is expressed throughout myeloid development, resulting in a broad distribution among neutrophil granules
TNF‐related apoptosis‐inducing ligand (TRAIL) is expressed throughout myeloid development, resulting in a broad distribution among neutrophil granules
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DOI：
10.1189/jlb.0707452
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发表时间：
2008-03
影响因子：
5.5
通讯作者：
M. Simons;K. Leidal;W. Nauseef;T. Griffith
中科院分区：
文献类型：
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作者：
M. Simons;K. Leidal;W. Nauseef;T. Griffith
TRAIL在多种肿瘤细胞中诱导凋亡。我们的实验室发现，人类中性粒细胞含有预制TRAIL的细胞内储库，其在用牛分枝杆菌卡介苗刺激后释放。在这项研究中，我们研究了新鲜分离的中性粒细胞中TRAIL的亚细胞分布。通过亚细胞分级分离神经元颗粒、分泌囊泡（SV）和质膜囊泡，然后进行自由流电泳，并通过ELISA和免疫印迹进行检查。在所有膜结合级分中发现TRAIL，其中在富含嗜天青颗粒（AG）和SV的级分中含量最高。免疫荧光共聚焦显微镜显示TRAIL与髓过氧化物酶（MPO）、乳铁蛋白（LF）和白蛋白（AG、特异性颗粒和SV的各自标记物）独立共定位。免疫透射电镜显示TRAIL与MPO、白蛋白共定位于细胞内。我们检测了用二甲基甲酰胺诱导的PLB-985细胞和用G-CSF处理的CD 34阳性干细胞中的TRAIL表达。定量RT-PCR分析表明，TRAIL在发育的每个阶段都有表达，而MPO和LF仅在分化过程中的不同时间表达。总的来说，这些研究结果表明，TRAIL在整个中性粒细胞发育过程中表达，导致不同颗粒亚型之间的广泛分布。
TRAIL induces apoptosis in a variety of tumor cells. Our laboratory found that human neutrophils contain an intracellular reservoir of prefabricated TRAIL that is released after stimulation with Mycobacterium bovis bacillus Calmette‐Guérin. In this study, we examined the subcellular distribution of TRAIL in freshly isolated neutrophils. Neutrophil granules, secretory vesicles (SV), and plasma membrane vesicles were isolated by subcellular fractionation, followed by free‐flow electrophoresis, and examined by ELISA and immunoblot. TRAIL was found in all membrane‐bound fractions with the highest amounts in the fractions enriched in azurophilic granule (AG) and SV. Immunofluorescence confocal microscopy showed that TRAIL colocalized independently with myeloperoxidase (MPO), lactoferrin (LF), and albumin, respective markers of AG, specific granules, and SV. Furthermore, immunotransmission electron microscopy demonstrated that TRAIL colocalized intracellularly with MPO and albumin. We examined TRAIL expression in PLB‐985 cells induced with dimethylformamide and in CD34‐positive stem cells treated with G‐CSF. Quantitative RT‐PCR analysis showed that TRAIL was expressed in each stage of development, whereas MPO and LF were only expressed at distinct times during differentiation. Collectively, these findings suggest that TRAIL is expressed throughout neutrophil development, resulting in a broad distribution among different granule subtypes.