Identification of a novel and highly potent eosinophil chemotactic lipid in human eosinophils treated with arachidonic acid.
Identification of a novel and highly potent eosinophil chemotactic lipid in human eosinophils treated with arachidonic acid.
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在用花生四烯酸处理的人嗜酸性粒细胞中鉴定出一种新型高效的嗜酸性粒细胞趋化脂质。
DOI：
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发表时间：
1990
影响因子：
4.4
通讯作者：
E. Christophers
中科院分区：
文献类型：
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作者：
E. Morita;J. Schröder;E. Christophers
纯化的人嗜酸性粒细胞产生嗜酸性粒细胞趋化脂质（ECL），当与花生四烯酸孵育时，没有任何刺激。孵育上清液的反相HPLC显示主要的脂质样嗜酸性粒细胞趋化活性，在含有5（S），15（S）二羟基-6，13-反式-8，11-顺式-二十碳四烯酸（5，15-DiHETE）以及8，15-二羟基二十碳四烯酸的峰中洗脱。为了进一步表征ECL，测试了一些真实的二羟基二十碳四烯酸的嗜酸性粒细胞趋化活性。然而，仅发现5，15-DiHETE以及8（S），15（S）-二羟基-5，11-顺式-9，13-反式-二十碳四烯酸是人嗜酸性粒细胞的趋化因子，其ED 50分别接近0.3 μ M和1.5 μ M。ECL制剂中高滴度嗜酸性粒细胞趋化活性的存在让我们寻找具有更高比活性的污染ECL。通过使用不同的反相HPLC系统，可以从高效ECL中分离5，15-DiHETE以及8（R），15（S）-二羟基-5，11-顺式-9，13-反式-二十碳四烯酸。通过使用直相HPLC对该ECL进行最终纯化，在260 nm处产生单一吸收峰，其UV光谱不同于已知的嗜酸性粒细胞趋化因子，表明嗜酸性粒细胞趋化脂质的新型。已发现纯化ECL的嗜酸性粒细胞趋化活性与血小板活化因子（迄今为止已知的最有效的趋化因子）的趋化活性相似，无论是迁移细胞的数量还是ED 50。交叉脱敏实验与ECL，白三烯B4，和血小板活化因子揭示了存在一个单独的ECL受体的嗜酸性粒细胞。由应答细胞自身产生的有效ECL支持存在嗜酸性粒细胞积累的自我维持机制的观点。
Purified human eosinophils generate eosinophil chemotactic lipids (ECL), when incubated with arachidonic acid without any stimulus. Reversed phase HPLC of incubation supernatants revealed major lipid-like eosinophil chemotactic activity eluting in a peak containing 5(S), 15(S)dihydroxy-6,13-trans-8,11-cis-eicosatetraenoic acid (5,15-DiHETE) as well as a 8,15-dihydroxyeicosatetraenoic acid. For further characterization of the ECL, some authentic dihydroxyeicosatetraenoic acids were tested for eosinophil chemotactic activity. Only 5,15-DiHETE as well as 8(S), 15(S)-dihydroxy-5,11-cis-9,13-trans-eicosatetraenoic acid were found to be chemotaxins for human eosinophils, however, with an ED50 near 0.3 microM and 1.5 microM, respectively. The presence of high titer eosinophil chemotactic activity in ECL preparations let us look for a contaminating ECL with higher specific activity. By using a different reversed phase HPLC-system 5,15-DiHETE as well as 8(R), 15(S)-dihydroxy-5,11-cis-9,13-trans-eicosatetraenoic acid could be separated from a highly potent ECL. Final purification of this ECL by the use of straight phase HPLC resulted in a single at 260 nm absorbing peak giving an UV spectrum different from that known for eosinophil chemotactic factors indicating a novel type of eosinophil chemotactic lipid. Eosinophil chemotactic activity of purified ECL has been found to be similar to that seen for platelet-activating factor, the most potent chemotaxin so far known, either in the number of migrating cells or the ED50. Cross-desensitization experiments with ECL, leukotriene B4, and platelet-activating factor revealed the existence of a separate ECL receptor on eosinophils. The production of potent ECL by the responder cells themselves supports the idea that there exists a self-sustaining mechanism of eosinophil accumulation.