Amphiregulin Production by Human Eosinophils
Amphiregulin Production by Human Eosinophils
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DOI：
10.1159/000210652
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发表时间：
2009-01-01
影响因子：
2.8
通讯作者：
Saito, Hirohisa
中科院分区：
文献类型：
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作者：
Matsumoto, Kenji;Fukuda, Shuhei;Saito, Hirohisa
背景：双调节蛋白（AREG）在乳腺发育、对线虫感染的免疫反应和肺粘膜生成中起着关键作用。由于在所有这些组织部位都显示出显著的嗜酸性粒细胞浸润，我们在体外检测了人类嗜酸性粒细胞产生的AREG。方法：采用Ficoll和抗体包被免疫磁珠，从正常或轻度过敏患者外周血中纯化嗜酸性粒细胞和其他血细胞。嗜酸性粒细胞在10 ng/ml粒细胞-巨噬细胞集落刺激因子（GM-CSF）、10 ng/ml IL-5、100 ng/ml ifn - γ或固定化分泌IgA的条件下培养。培养6 h后提取总mRNA，用芯片和RT-PCR检测mRNA表达量。采用ELISA法测定培养48h后上清液和细胞裂解液中AREG的浓度。结果：AREG mRNA在新鲜嗜酸性粒细胞中组成性表达，且在嗜酸性粒细胞中的表达水平高于其他类型血细胞。在GM-CSF和IL- 5的刺激下AREG mRNA显著升高，而ifn - γ的刺激下AREG mRNA无显著升高。地塞米松可增强gm - csf刺激下嗜酸性粒细胞中AREG mRNA的表达。新鲜嗜酸性粒细胞不含AREG蛋白。当嗜酸性粒细胞被GM-CSF刺激时，AREG被显著释放到上清中，而ifn - γ或固定化分泌IgA则没有。结论：我们的数据表明，嗜酸性粒细胞在体内产生和释放AREG，并参与一些生理和病理条件。版权所有(c) 2009 S. Karger AG，巴塞尔
Background: Amphiregulin (AREG) plays critical roles in mammary gland development, immune responses against nematode infection, and mucous production in the lung. Since remarkable eosinophil infiltration has been shown at all of these tissue sites, we examined AREG production by human eosinophils in vitro. Methods: Using Ficoll and antibody-coated immunomagnetic beads, eosinophils and other blood cells were purified from peripheral blood samples obtained from normal or mild allergic patients. Eosinophils were cultured in the presence of 10 ng/ml of granulocyte-macrophage colony-stimulating factor (GM-CSF), 10 ng/ml of IL-5, 100 ng/ml of IFN-gamma or immobilized secretory IgA. Total mRNA was extracted after 6 h of culture, and mRNA expression was measured using a microarray and RT-PCR. The AREG concentration in the supernatant and cell lysate after 48 h of culture was measured using ELISA. Results: AREG mRNA was constitutively expressed in fresh eosinophils, and the expression level in the eosinophils was higher than that in other types of blood cells. AREG mRNA increased significantly upon stimulation with GM-CSF and IL- 5 but not with IFN-gamma. AREG mRNA expression in GM-CSF-stimulated eosinophils was enhanced by dexamethasone. Fresh eosinophils did not contain AREG protein. AREG was significantly released into the supernatant when the eosinophils were stimulated with GM-CSF, but not with IFN-gamma or immobilized secretory IgA. Conclusion: Our data suggested that eosinophils produce and release AREG, and participate in some physiological and pathological conditions in vivo. Copyright (c) 2009 S. Karger AG, Basel