Egger D, Geuenich S, Denzlinger C, Schmitt E, Mailhammer R, Ehrenreich H, Dörmer P, Hültner L
GSF-Institute for Experimental Hematology, Munich, Germany.
J Immunol. 1995 Feb 15;154(4):1830-7.
It has previously been shown that mouse bone marrow-derived mast cells (BMMC) synthesize and secrete endothelin-1 (ET-1) and express ETA-type endothelin receptors (ETA-R). The study presented here was designed to elucidate the influence of different cytokine conditions for cellular differentiation and maturation on the ability of primary mouse BMMC to respond to exogenous ET-1. BMMC were grown for 2 wk in IL-3 alone and then cultured for 2 to 3 wk with kit ligand (KL) and/or IL-3 in the presence or absence of IL-4. ET-1 induced a very rapid (< or = 1 min) and dose-dependent release of histamine and serotonin from BMMC cultured in the presence of both IL-3 and IL-4. The effect of ET-1 was quantitatively comparable with IgE/Ag-induced mediator release and comprised up to 20% and 16% of total cellular histamine and serotonin, respectively. In BMMC grown with KL or KL plus IL-3, a substantial effect of ET-1 on amine release was only observed when IL-4 had been included in the culture medium. These IL-4 effects could not be observed if BMMC grown in IL-3 and/or KL were preincubated for 1 or 24 h with IL-4 before activation with ET-1, suggesting that a differentiation process rather than a functional priming effect had been initiated by IL-4. In BMMC, the histamine and serotonin release induced by ET-1 (10(-6) M) was inhibited by an ETA-R-specific antagonist (cyclic [D-Asp-Pro-D-Val-Leu-D-Trp]) in a dose-dependent manner, with complete inhibition at an antagonist concentration of 10(-8) M. ET-1 stimulated leukotriene C4 biosynthesis up to 4.5-fold in BMMC cultured in the presence of IL-4. No such ET-1 effect was observed in BMMC cultured in media containing IL-3, KL, or a combination of both cytokines. Peritoneal cells (containing 2 to 3% serosal mast cells) obtained from BALB/c mice released 87 +/- 2% of histamine within 1 min after challenge with ET-1. Our results demonstrate that ET-1 can directly act as a histamine and serotonin secretagogue and as a stimulator of leukotriene C4 production in mast cells. IL-4 appears to be critically involved in the differentiation of immature mast cell precursors to an ET-1-reactive phenotype.(ABSTRACT TRUNCATED AT 400 WORDS)
先前的研究表明，小鼠骨髓来源的肥大细胞（BMMC）能合成并分泌内皮素-1（ET-1），并表达ETA型内皮素受体（ETA-R）。本文所呈现的研究旨在阐明不同细胞因子条件对细胞分化和成熟的影响，以及对原代小鼠BMMC对外源性ET-1反应能力的影响。BMMC先在单独的IL-3中培养2周，然后在有或无IL-4的情况下，与kit配体（KL）和/或IL-3一起培养2至3周。ET-1能在IL-3和IL-4共同存在的情况下，非常迅速地（≤1分钟）且呈剂量依赖性地诱导BMMC释放组胺和5-羟色胺。ET-1的作用在数量上与IgE/抗原诱导的介质释放相当，分别占细胞总组胺和5-羟色胺的20%和16%。在与KL或KL加IL-3一起培养的BMMC中，只有当培养基中加入IL-4时，ET-1对胺释放才有显著作用。如果在IL-3和/或KL中培养的BMMC在用ET-1激活前用IL-4预孵育1或24小时，就观察不到这些IL-4的作用，这表明IL-4启动的是一个分化过程，而非功能性的致敏作用。在BMMC中，ETA-R特异性拮抗剂（环[D-天冬氨酸-脯氨酸-D-缬氨酸-亮氨酸-D-色氨酸]）能以剂量依赖性方式抑制ET-1（10⁻⁶ M）诱导的组胺和5-羟色胺释放，在拮抗剂浓度为10⁻⁸ M时完全抑制。在有IL-4存在的情况下培养的BMMC中，ET-1能刺激白三烯C4的生物合成增加至4.5倍。在含有IL-3、KL或这两种细胞因子组合的培养基中培养的BMMC中未观察到这种ET-1的作用。从BALB/c小鼠获得的腹腔细胞（含2%至3%的浆膜肥大细胞）在用ET-1刺激后1分钟内释放了87±2%的组胺。我们的结果表明，ET-1可直接作为组胺和5-羟色胺的促分泌剂，以及肥大细胞中白三烯C4产生的刺激物。IL-4似乎在未成熟肥大细胞前体向ET-1反应性表型的分化过程中起关键作用。（摘要截短至400字）
