Tropism, replication competence, and innate immune responses of influenza virus: an analysis of human airway organoids and ex-vivo bronchus cultures
Tropism, replication competence, and innate immune responses of influenza virus: an analysis of human airway organoids and ex-vivo bronchus cultures
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DOI：
10.1016/s2213-2600(18)30236-4
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发表时间：
2018-11-01
影响因子：
76.2
通讯作者：
Chan, Michael C. W.
中科院分区：
文献类型：
--
作者：
Hui, Kenrie P. Y.;Ching, Rachel H. H.;Chan, Michael C. W.
人类气道类器官是来源于干细胞的三维培养物，其在离体条件下自组织形成所谓的迷你气道。这些培养物的细胞形态在生理上类似于人气道，纤毛、杯状细胞和dub细胞面向内腔，基底细胞位于外层。本研究的目的是比较人和禽流感病毒株在离体人支气管和人气道类器官中引起的复制能力、组织嗜性和宿主反应。方法在2016年9月29日至2017年1月4日期间，我们得到了前-人支气管的体内培养物和来自肺干细胞的培养的人气道类器官，所述肺干细胞从作为常规临床试验的一部分移除的人肺组织获得。护理在香港大学玛丽医院心胸外科接受手术切除的病人。我们比较了从人类分离的A型禽流感病毒的病毒复制能力、组织嗜性、细胞因子和趋化因子诱导（Sh2/H7N9，H5N1/483，H5N6/39715），和人H1N1 pdm/415742在气道类器官和前结果人类和禽流感A病毒在人类气道类器官中的病毒嗜性和复制动力学模拟了在人支气管外植体的离体培养物。在气道类器官和支气管外植体中，甲型流感H1N1亚型（H1N1）和甲型禽流感H7N9病毒复制的滴度显著高于高致病性禽流感（HPAI）H5 N1，而HPAI H5 N6复制是中等的。H1N1、H7N9和H5 N6病毒感染气道类器官和支气管外植体中的纤毛细胞和杯状细胞，但不感染基底细胞。细胞因子、白细胞介素6和干扰素β的表达，以及活化调节的趋化因子、正常T细胞表达和分泌，在感染HPAI H5 N1病毒的人气道类器官中显著高于H1N1 pdm/415742、Sh 2/H7N9和H5 N6/39715病毒，单核细胞趋化蛋白-1在HPAI H5 N1病毒感染的人类器官中的表达显著高于H1N1 pdm/415742和Sh 2/人类气道类器官培养物提供的结果与在人类离体支气管培养物中观察到的结果相当，因此提供了用于研究病毒嗜性和复制能力的替代生理学相关实验模型，其可用于评估动物流感病毒的大流行威胁。版权所有（C）2018 Elsevier Ltd.保留所有权利。
Background Human airway organoids are three-dimensional cultures derived from stem cells, which self-organise in ex-vivo conditions to form so-called mini-airways. The cellular morphology of these cultures is physiologically similar to the human airway, with cilia, goblet cells, and dub cells facing the inner lumen and basal cells situated at the outer layer. The aim of this study was to compare replication competence, tissue tropism, and host responses elicited by human and avian strains of influenza A virus in ex-vivo human bronchus and human airway organoids.Methods Between Sept 29, 2016, and Jan 4, 2017, we obtained ex-vivo cultures of the human bronchus and cultured human airway organoids from lung stem cells obtained from human lung tissues removed as part of the routine clinical care of patients undergoing surgical resection at the Department of Cardiothoracic Surgery, University of Hong Kong, Queen Mary Hospital, Hong Kong. We compared viral replication competence, tissue tropism, and cytokine and chemokine induction of avian influenza A viruses isolated from humans (Sh2/H7N9, H5N1/483, H5N6/39715), and human H1N1pdm/415742 in airway organoids and ex-vivo bronchus explant cultures.Findings Virus tropism and replication kinetics of human and avian influenza A viruses in human airway organoids mimicked those found in ex-vivo cultures of human bronchus explants. In both airway organoids and bronchus explants, influenza A H1N1 subtype (H1N1) and avian influenza A H7N9 viruses replicated to significantly higher titres than did the highly pathogenic avian influenza (HPAI) H5N1, whereas HPAI H5N6 replication was moderate. H1N1, H7N9, and H5N6 viruses infected ciliated cells and goblet cells, but not basal cells in both airway organoids and bronchus explants. The expression of cytokines, interleukin 6, and interferon beta, and the chemokine regulated-on-activation, normal T-cell expressed and secreted, was significantly higher in human airway organoids infected with HPAI H5N1 virus than H1N1pdm/415742, Sh2/H7N9, and H5N6/39715 viruses, and the expression of monocyte chemoattractant protein-1 was significantly higher in human organoids infected with HPAI H5N1 virus than H1N1pdm/415742 and Sh2/H7N9 viruses.Interpretation Human airway organoid cultures provided results that were comparable to those observed in human ex-vivo bronchus cultures, and thus provide an alternative physiologically relevant experimental model for investigating virus tropism and replication competence that could be used to assess the pandemic threat of animal influenza viruses. Copyright (C) 2018 Elsevier Ltd. All rights reserved.