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首页> 外文期刊>Allergy >A critical role for vesicle-associated membrane protein-7 in exocytosis from human eosinophils and neutrophils.
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A critical role for vesicle-associated membrane protein-7 in exocytosis from human eosinophils and neutrophils.

机译：囊泡相关膜蛋白7在人类嗜酸性粒细胞和嗜中性粒细胞胞吐作用中的关键作用。

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Background: Granulocyte exocytosis is proposed to be critically dependent on the interaction of soluble N-ethylmaleimide-sensitive factor attachment protein (SNAP) receptors (SNAREs) located on granules/vesicles (v-SNAREs) and plasma membrane (t-SNAREs). Previous studies indicated that the v-SNARE, vesicle-associated membrane protein (VAMP)-2, as well as t-SNAREs (SNAP-23, syntaxin-4 and -6) are implicated in exocytosis from human granulocytes. Vesicle-associated membrane proteins-7 and -8 have been implicated in endosome/lysosome trafficking, however, their role in granulocyte exocytosis remains obscure. Objective: We sought to investigate the expression and functional role of SNARE isoforms in the secretion of different granule-derived mediators in human eosinophils and neutrophils. Methods: The expression of SNAREs was determined by subcellular fractionation and flow cytometry. SNARE-specific antibodies were examined for their ability to impair mediator release from permeabilized eosinophils and neutrophils. Results: Vesicle-associated membrane proteins-7 and -8 were localized to granule and membrane-enriched fractions in eosinophils and neutrophils, whereas syntaxin-6 was not detectable. In permeabilized cells, anti-VAMP-7, but not anti-VAMP-8, antibody impaired the secretion of all mediators examined (in eosinophils, eosinophil peroxidase and eosinophil-derived neurotoxin; in neutrophils, myeloperoxidase, lactoferrin and matrix metalloprotease-9) in a dose-dependent manner. In contrast, anti-VAMP-2 modestly and selectively impaired secretion from small granules and vesicles. Syntaxin-4, but not syntaxin-6, was found to interact with SNAP-23 and was partially involved in mediator secretion from multiple compartments. Conclusion: Our observations indicate for the first time a critical role for VAMP-7 in both eosinophil and neutrophil mediator release.
机译：背景：粒细胞的胞吐作用被认为主要取决于位于颗粒/小泡（v-SNARE）和质膜（t-SNARE）上的可溶性N-乙基马来酰亚胺敏感因子附着蛋白（SNAP）受体（SNARE）的相互作用。先前的研究表明，v-SNARE，囊泡相关膜蛋白（VAMP）-2和t-SNARE（SNAP-23，syntaxin-4和-6）与人类粒细胞的胞吐作用有关。囊泡相关的膜蛋白7和-8与内体/溶酶体运输有关，但是，它们在粒细胞胞吐作用中的作用仍然不清楚。目的：我们试图研究SNARE同工型在人类嗜酸性粒细胞和嗜中性粒细胞分泌不同颗粒的介质中的表达及其功能。方法：通过亚细胞分级分离和流式细胞仪检测SNAREs的表达。检测了SNARE特异性抗体的能力，以防止其从透化的嗜酸性粒细胞和嗜中性粒细胞中释放介质。结果：囊泡相关的膜蛋白7和-8定位于嗜酸性粒细胞和嗜中性粒细胞的颗粒和膜富集部分，而无法检测到syntaxin-6。在通透性细胞中，抗​​VAMP-7抗体而非抗VAMP-8抗体损害了所有检查的介质的分泌（在嗜酸性粒细胞，嗜酸性粒细胞过氧化物酶和嗜酸性粒细胞衍生的神经毒素中;在中性粒细胞中，髓过氧化物酶，乳铁蛋白和基质金属蛋白酶-9）以剂量依赖的方式。相反，抗VAMP-2适度和选择性地损害了小颗粒和囊泡的分泌。发现Syntaxin-4，而不是syntaxin-6，与SNAP-23相互作用，并部分参与多个区室的介体分泌。结论：我们的观察首次表明VAMP-7在嗜酸性粒细胞和嗜中性粒细胞介质释放中均起关键作用。

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