上皮外泌体接触蛋白 1 促进在哮喘中单核细胞衍生的树突状细胞主导的 T 细胞反应
2021/06/18
摘要
背景：外泌体已成为细胞通讯的重要参与者。然而，气道上皮细胞（AEC）产生的外泌体是否参与哮喘的发展仍然未知。
目的：我们的目标是表征 AEC 分泌的外泌体和可能有助于AEC外泌体在树突状细胞(DC)主导气道过敏模型中的促炎作用的潜在功能蛋白，并确认它们在哮喘患者中的临床意义.
方法：用源自屋尘螨(HDM)刺激的AEC(HDM-AEC-EXO)或由HDM和/或contactin-1(CNTN1) 引发的单核细胞衍生的DC的外泌体处理小鼠。通过流式细胞术测定肺中DC的数量。对纯化的HDM-AECEXO进行蛋白质组学分析。CNTN1小干扰RNA旨在探测其在气道过敏中的作用，并使用g-分泌酶抑制剂来确定Notch通路的参与。
结果：HDM-AEC-EXO促进细胞培养和小鼠中单核细胞衍生的DC的募集、增殖、迁移和激活。外泌体中的CNTN1是哮喘病理学的关键参与者。RNA干扰介导的沉默和药物抑制剂表明 Notch2受体是传递CNTN1信号以激活TH2细胞/TH17细胞免疫反应所必需的。对哮喘患者的研究也支持在细胞和小鼠模型中观察到的 CNTN1-Notch2 轴的存在。
结论：这项研究的发现揭示了CNTN1在通过外泌体分泌介导的哮喘发病机制中的新作用，表明了治疗过敏性气道炎症的潜在策略。
Epithelial exosomal contactin-1 promotes monocyte-derived dendritic cell–dominant T-cell responses in asthma
Zhang M, Yu Q, Tang W, et al.
Abstract
BACKGROUND: Exosomes have emerged as a vital player in cell cell communication; however, whether airway epithelial cell (AEC)-generated exosomes participate in asthma development remains unknown.
OBJECTIVE:Our aims were to characterize the AEC-secreted exosomes and the potentially functional protein(s) that may contribute to the proinflammatory effects of AEC exosomes in the dendritic cell (DC)-dominant airway allergic models and to confirm their clinical significance in patients with asthma.
METHODS: Mice were treated with exosomes derived from house dust mite (HDM)-stimulated AECs (HDM-AEC-EXOs) or monocyte-derived DCs primed by HDM and/or contactin-1 (CNTN1). The numbers of DCs in the lung were determined by flow cytometry. Proteomic analysis of purified HDM-AECEXOs was performed. CNTN1 small interfering RNA was designed to probe its role in airway allergy, and g-secretase inhibitor was used to determine involvement of the Notch pathway.
RESULTS:HDM-AEC-EXOs facilitate the recruitment, proliferation, migration, and activation of monocyte-derived DCs in cell culture and in mice. CNTN1 in exosomes is a critical player in asthma pathology. RNA interference–mediated silencing and pharmaceutical inhibitors characterize Notch2 receptor as necessary for relaying the CNTN1 signal to activate TH2 cell/TH17 cell immune response. Studies of patients with asthma also support existence of the CNTN1-Notch2 axis that has been observed in cell and mouse models.
CONCLUSIONS:This study’s findings reveal a novel role for CNTN1 in asthma pathogenesis mediated through exosome secretion, indicating a potential strategy for the treatment of allergic airway inflammation.
背景：外泌体已成为细胞通讯的重要参与者。然而，气道上皮细胞（AEC）产生的外泌体是否参与哮喘的发展仍然未知。
目的：我们的目标是表征 AEC 分泌的外泌体和可能有助于AEC外泌体在树突状细胞(DC)主导气道过敏模型中的促炎作用的潜在功能蛋白，并确认它们在哮喘患者中的临床意义.
方法：用源自屋尘螨(HDM)刺激的AEC(HDM-AEC-EXO)或由HDM和/或contactin-1(CNTN1) 引发的单核细胞衍生的DC的外泌体处理小鼠。通过流式细胞术测定肺中DC的数量。对纯化的HDM-AECEXO进行蛋白质组学分析。CNTN1小干扰RNA旨在探测其在气道过敏中的作用，并使用g-分泌酶抑制剂来确定Notch通路的参与。
结果：HDM-AEC-EXO促进细胞培养和小鼠中单核细胞衍生的DC的募集、增殖、迁移和激活。外泌体中的CNTN1是哮喘病理学的关键参与者。RNA干扰介导的沉默和药物抑制剂表明 Notch2受体是传递CNTN1信号以激活TH2细胞/TH17细胞免疫反应所必需的。对哮喘患者的研究也支持在细胞和小鼠模型中观察到的 CNTN1-Notch2 轴的存在。
结论：这项研究的发现揭示了CNTN1在通过外泌体分泌介导的哮喘发病机制中的新作用，表明了治疗过敏性气道炎症的潜在策略。
（中日友好医院呼吸与危重症医学科 李春晓 摘译 林江涛 审校）
（J Allergy Clin Immunol, 2021）
（J Allergy Clin Immunol, 2021）
Epithelial exosomal contactin-1 promotes monocyte-derived dendritic cell–dominant T-cell responses in asthma
Zhang M, Yu Q, Tang W, et al.
Abstract
BACKGROUND: Exosomes have emerged as a vital player in cell cell communication; however, whether airway epithelial cell (AEC)-generated exosomes participate in asthma development remains unknown.
OBJECTIVE:Our aims were to characterize the AEC-secreted exosomes and the potentially functional protein(s) that may contribute to the proinflammatory effects of AEC exosomes in the dendritic cell (DC)-dominant airway allergic models and to confirm their clinical significance in patients with asthma.
METHODS: Mice were treated with exosomes derived from house dust mite (HDM)-stimulated AECs (HDM-AEC-EXOs) or monocyte-derived DCs primed by HDM and/or contactin-1 (CNTN1). The numbers of DCs in the lung were determined by flow cytometry. Proteomic analysis of purified HDM-AECEXOs was performed. CNTN1 small interfering RNA was designed to probe its role in airway allergy, and g-secretase inhibitor was used to determine involvement of the Notch pathway.
RESULTS:HDM-AEC-EXOs facilitate the recruitment, proliferation, migration, and activation of monocyte-derived DCs in cell culture and in mice. CNTN1 in exosomes is a critical player in asthma pathology. RNA interference–mediated silencing and pharmaceutical inhibitors characterize Notch2 receptor as necessary for relaying the CNTN1 signal to activate TH2 cell/TH17 cell immune response. Studies of patients with asthma also support existence of the CNTN1-Notch2 axis that has been observed in cell and mouse models.
CONCLUSIONS:This study’s findings reveal a novel role for CNTN1 in asthma pathogenesis mediated through exosome secretion, indicating a potential strategy for the treatment of allergic airway inflammation.
