Identification, purification, and characterization of a mast cell-associated cytolytic factor related to tumor necrosis factor.
Identification, purification, and characterization of a mast cell-associated cytolytic factor related to tumor necrosis factor.
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DOI：
10.1073/pnas.84.24.9175
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发表时间：
1987-12
影响因子：
11.1
通讯作者：
John Ding-E Young;Chau‐Ching Liu;G. Butler;Z. A. Cohn;Stephen J. Galli
中科院分区：
文献类型：
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作者：
John Ding-E Young;Chau‐Ching Liu;G. Butler;Z. A. Cohn;Stephen J. Galli
研究了肥大细胞和肥大细胞衍生因子在自然细胞毒性反应中的作用。培养的和新鲜分离的鼠肥大细胞在18小时活力测定中显示对WEHI-164和YAC-1靶标具有细胞毒性，但在4小时测定中没有。在这里，我们描述了一种细胞毒性因子在小鼠肥大细胞，是免疫相关的肿瘤坏死因子（TNF）。这种TNF样因子以缓慢的时间过程裂解WEHI-164细胞，需要16-20小时才能完成裂解反应。特异于人和鼠TNF和人α-光敏素的抗体部分阻断WEHI-164细胞的肥大细胞溶解。这些抗体在免疫印迹上与50 kDa的一条主要肥大细胞蛋白带反应。免疫印迹分析表明，在克隆和未克隆培养的小鼠肥大细胞，并在成熟的“结缔组织型”肥大细胞新鲜纯化的大鼠或小鼠腹腔中的这个因素。该因子的量在已经用佛波酯/伴刀豆球蛋白A或细菌脂多糖的组合刺激的细胞中大大增强。用Percoll梯度对肥大细胞进行亚细胞分级分离分析，发现两个TNF相关细胞毒活性池与游离胞质物质和颗粒组分相关。与细胞毒性T淋巴细胞和自然杀伤细胞相反，肥大细胞的颗粒富集组分不含任何溶血活性。肥大细胞颗粒中TNF样分子的定位可能在细胞表面刺激和脱粒后将该介质快速递送至靶细胞膜中发挥战略性作用。
The role of mast cells and mast-cell-derived factors in natural cytotoxic reactions was investigated. Cultured and freshly isolated murine mast cells are shown to be cytotoxic to WEHI-164 and YAC-1 targets in 18-hr viability assays but not in 4-hr assays. Here, we describe a cytotoxic factor in murine mast cells that is immunologically related to tumor necrosis factor (TNF). This TNF-like factor lyses WEHI-164 cells with a slow time course requiring 16-20 hr for the lytic reaction to complete. Antibodies specific for human and murine TNF and human lymphotoxin partially block mast cell lysis of WEHI-164 cells. These antibodies react on immunoblots with one major mast cell protein band of 50 kDa. Immunoblot analysis shows this factor in cloned and uncloned cultured mouse mast cells and in mature "connective tissue-type" mast cells freshly purified from rat or mouse peritoneal cavities. The amount of this factor is greatly enhanced in cells that have been stimulated with a combination of phorbol ester/concanavalin A or bacterial lipopolysaccharide. Subcellular fractionation analysis of mast cells with Percoll gradients reveals two pools of TNF-related cytotoxic activity that are associated with free cytosolic material and granule fractions. In contrast to cytotoxic T lymphocytes and natural killer cells, granule-enriched fractions of mast cells do not contain any hemolytic activity. The localization of the TNF-like molecule in mast cell granules may play a strategical role in the rapid delivery of this mediator to the target cell membrane following cell surface stimulation and degranulation.