Expression of the chemokine receptor CCR3 on human mast cells
Expression of the chemokine receptor CCR3 on human mast cells
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DOI：
10.1159/000053694
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发表时间：
2001-01-01
影响因子：
2.8
通讯作者：
Romagnani, P
中科院分区：
文献类型：
--
作者：
de Paulis, A;Annunziato, F;Romagnani, P
背景：本研究的目的是调查人类肥大细胞是否表达功能活性 CCR3 受体，这些受体由 CC 趋化因子激活。这些配体包括 CCR3 选择性趋化因子 eotaxin 和 eotaxin-2 以及更混杂的 CC 趋化因子 MCP-4、MCP-3、MCP-2 和 RANTES。方法：对皮肤、肠道和肺部标本进行免疫组织化学分析。通过使用具有两种不同底物的抗生物素蛋白-生物素-过氧化物酶系统，用抗CCR3和抗类胰蛋白酶以及抗CCR3和抗糜酶抗体(Ab)进行双重免疫染色。通过逆流淘洗和不连续 Percoll 密度梯度从人肺实质 (HLMC) 中分离和纯化肥大细胞。使用单克隆抗体抗 CCR3 (7B11) 对 HLMC 表面 CCR3 表达进行流式细胞术分析。 H LMC 的功能激活通过细胞响应嗜酸细胞趋化因子而释放组胺和/或迁移的能力来验证。结果：在皮肤和肠粘膜下层中发现了高百分比（>70%）的显示 CCR3 表达的类胰蛋白酶阳性细胞，而在肠粘膜和肺间质中发现了低得多的百分比（小于或等于 20%）。 Eotaxin (1-100 nM) 既不诱导 HLMC 释放组胺，也不增强抗 IgE 诱导的组胺释放。相反，eotaxin (10-100 nM) 和 RANTES (10-100 nM) 在体外诱导 HLMC 趋化性。在装入趋化室之前，将 HLMC 与抗 CCR3 抗体 (5 mug/ml) 预温育，消除了由嗜酸细胞趋化因子引起的趋化性。抗CCR3和抗糜酶抗体的双重免疫染色显示，所检查的各种人体组织中绝大多数表达CCR3的肥大细胞是类胰蛋白酶-糜酶双阳性。结论：这些结果表明 CCR3 在人类肥大细胞上表达，并且这些细胞被 CCR3 结合趋化因子吸引。版权所有 (C) 2001 S. Karger AG，巴塞尔。
Background: The aim of this study was to investigate whether human mast cells express functional active CCR3 receptors, which are activated by CC chemokines. These ligands include the CCR3-selective chemokines eotaxin and eotaxin-2 and the more promiscuous CC chemokines, MCP-4, MCP-3, MCP-2 and RANTES. Methods: Immunohistochemical analysis was performed on skin, gut and lung specimens. Double immunostaining was performed with anti-CCR3 and antitryptase, and anti-CCR3 and antichymase antibody (Ab) by using the avidin-biotin-peroxidase system with two different substrates. Mast ce[is were isolated and purified from human lung parenchyma (HLMC) by countercurrent elutriation followed by discontinuous Percoll density gradient. Flow-cytometric analysis of HLMC surface CCR3 expression was performed with the monoclonal Ab anti-CCR3 (7B11). Functional activation of H LMC was verified by th e ability of cells to release histamine and/or migrate in response to eotaxin. Results: High percentages (>70%) of tryptase-positive cells showing CCR3 expression were found in the skin and in the intestinal submucosa, whereas much lower percentages (less than or equal to 20%) were found in the intestinal mucosa and in the lung interstitium. Eotaxin (1-100 nM) neither induced histamine release from HLMC nor enhanced anti-IgE-induced histamine release. In contrast, eotaxin (10-100 nM) and RANTES (10-100 nM) induced HLMC chemotaxis in vitro. Preincubation of HLMC with antibody anti-CCR3 (5 mug/ml) before loading into the chemotaxis chamber abrogated chemotaxis elicited by eotaxin. Double immunostaining with anti-CCR3 and anti-chymase antibody showed that the vast majority of CCR3-expressing mast cells in the Various human tissues examined were tryptase-chymase double-positive. Conclusions: These results indicate that CCR3 is expressed on human mast cells and that these cells are attracted by CCR3-binding chemokines. Copyright (C) 2001 S. Karger AG, Basel.