Dynamics of leukotriene C production by macrophages.
Dynamics of leukotriene C production by macrophages.
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DOI：
10.1084/jem.152.5.1236
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发表时间：
1980-11-01
期刊：
影响因子：
--
通讯作者：
Cohn ZA
中科院分区：
文献类型：
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作者：
Rouzer CA;Scott WA;Hamill AL;Cohn ZA
建立了小鼠腹腔巨噬细胞体外产生低密度脂蛋白的放射化学分析方法。该方法包括用[5，6，8，9，11，12，14，15-3H]20：4标记培养的巨噬细胞，然后在所需的实验条件下刺激花生四烯酸(20：4)的释放。放射性标记白三烯C(LTC)通过萃取和硅酸层析从培养基中回收，产率为40%，完全保留了生物活性。由于这种LTC是放射化学纯的，LTC的释放量可以从样品中的放射性总量来估计。用放射分析法研究影响巨噬细胞合成LTC的参数表明，巨噬细胞合成LTC的时间进程及其与吞噬细胞刺激物(酵母多糖)剂量的关系与前列腺素(PG)的释放非常相似。LTC的释放也与PG相似，因为这两种代谢物都是由微小棒状杆菌诱导的巨噬细胞产生的，比常驻细胞产生的水平更低。最后，抗原-抗体复合体可刺激LTC的释放，但其最大释放水平低于酵母多糖。这里提供的数据与假设一致，即巨噬细胞在吞噬刺激下的刺激会导致可诱导的磷脂酶释放20：4。然后，环氧合酶和脂氧合酶竞争释放的20：4，导致PG、羟基二十碳四烯酸和LTC的产生。
A method for the radiochemical assay of LTC production by mouse peritoneal macrophages in vitro is presented. The method involves labeling macrophages in culture with [5,6,8,9,11,12,14,15-3H]20:4 followed by stimulation of arachidonic acid (20:4) release under the experimental conditions desired. Radiolabeled leukotriene C (LTC) is recovered from the culture medium by extraction and silicic acid chromatography in 40% yield with full retention of biological activity. Because this LTC is radiochemically pure, the quantity of LTC release may be estimated from the amount of radioactivity in the sample. Use of the radioassay to study parameters affecting LTC synthesis by macrophages indicated that the time course of LTC synthesis and its relationship to the dose of a phagocytic stimulus (zymosan) were very similar to those of prostaglandin (PG) release. LTC release was also similar to that of PG in that lower levels of both metabolites were produced by Corynebacterium parvum-elicited macrophages than by resident cells. Finally, LTC release was stimulated in response to a challenge with antigen-antibody complexes, but lower maximal levels were attained than those with zymosan. The data presented here are consistent with the hypothesis that challenge of macrophages with a phagocytic stimulus leads to the release of 20:4 by an inducible phospholipase. Cyclooxygenase and lipoxygenase then compete for the released 20:4, leading to the production of PG, hydroxyeicosatetraenoic acids, and LTC.