Human eosinophils constitutively express a unique serine protease, PRSS33
Human eosinophils constitutively express a unique serine protease, PRSS33
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DOI：
10.1016/j.alit.2017.01.001
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发表时间：
2017-07-01
影响因子：
6.8
通讯作者：
Matsumoto, Kenji
中科院分区：
文献类型：
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作者：
Toyama, Sumika;Okada, Naoko;Matsumoto, Kenji
背景资料：嗜酸性粒细胞通过产生多种颗粒蛋白、化学介质、细胞因子、趋化因子和蛋白酶等在哮喘尤其是气道重塑中发挥重要作用。然而，嗜酸性粒细胞的蛋白酶生产尚未完全了解。在本研究中，我们研究了嗜酸性粒细胞特异性蛋白酶/蛋白酶的生产通过transcriptome analysis.Methods：人嗜酸性粒细胞和其他细胞从外周血中纯化的密度梯度沉降和负/正选择使用免疫磁珠。通过微阵列分析、qPCR、ELISA、流式细胞术和免疫荧光染色在用嗜酸性粒细胞活化细胞因子和促分泌素刺激之前和之后评价嗜酸性粒细胞中的蛋白酶/蛋白酶表达和释放到上清液中。在暴露于重组丝氨酸蛋白酶33（PRSS 33）蛋白（rPRSS 33）后，通过qPCR测量人正常成纤维细胞中细胞外基质蛋白的mRNA，所述重组丝氨酸蛋白酶33蛋白（rPRSS 33）是用杆状病毒系统产生的。PRSS 33的表达最高，并且是嗜酸性粒细胞特异性的。PRSS 33 mRNA表达不受嗜酸性粒细胞活化细胞因子的影响。免疫荧光染色显示PRSS 33与嗜酸性粒细胞颗粒蛋白共定位。PRSS 33在嗜酸性粒细胞的培养上清液中未检测到，即使在用促分泌素刺激后，但其细胞表面表达增加。rPRSS 33刺激人成纤维细胞增加胶原和纤连蛋白mRNAs的表达，至少部分通过蛋白酶激活受体-2 activation.Conclusions：活化的嗜酸性粒细胞可能通过细胞表面表达PRSS 33诱导成纤维细胞细胞外基质蛋白合成，这将至少部分解释嗜酸性粒细胞在气道重塑中的作用。Copyright（C）2017，日本变态反应学会.制作和主办：Elsevier B. V.
Background: Eosinophils play important roles in asthma, especially airway remodeling, by producing various granule proteins, chemical mediators, cytokines, chemokines and proteases. However, protease production by eosinophils is not fully understood. In the present study, we investigated the production of eosinophil-specific proteases/proteinases by transcriptome analysis.Methods: Human eosinophils and other cells were purified from peripheral blood by density gradient sedimentation and negative/positive selections using immunomagnetic beads. Protease/proteinase expression in eosinophils and release into the supernatant were evaluated by microarray analysis, qPCR, ELISA, flow cytometry and immunofluorescence staining before and after stimulation with eosinophil-activating cytokines and secretagogues. mRNAs for extracellular matrix proteins in human normal fibroblasts were measured by qPCR after exposure to recombinant protease serine 33 (PRSS33) protein (rPRSS33), created with a baculovirus system.Results: Human eosinophils expressed relatively high levels of mRNA for metalloproteinase 25 (MMP25), a disintegrin and metalloprotease 8 (ADAM8), ADAM10, ADAM19 and PRSS33. Expression of PRSS33 was the highest and eosinophil-specific. PRSS33 mRNA expression was not affected by eosinophil-activating cytokines. Immunofluorescence staining showed that PRSS33 was co-localized with an eosinophil granule protein. PRSS33 was not detected in the culture supernatant of eosinophils even after stimulation with secretagogues, but its cell surface expression was increased. rPRSS33 stimulation of human fibroblasts increased expression of collagen and fibronectin mRNAs, at least in part via protease-activated receptor-2 activation.Conclusions: Activated eosinophils may induce fibroblast extracellular matrix protein synthesis via cell surface expression of PRSS33, which would at least partly explain eosinophils' role(s) in airway remodeling. Copyright (C) 2017, Japanese Society of Allergology. Production and hosting by Elsevier B.V.