Article(id=1297571058056455039, tenantId=1146029695717560320, journalId=1192105938417971205, issueId=1297570992835023717, articleNumber=null, orderNo=null, doi=10.13343/j.cnki.wsxb.20260154, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=research-article, receivedDate=1772035200000, receivedDateStr=2026-02-26, revisedDate=null, revisedDateStr=null, acceptedDate=1774886400000, acceptedDateStr=2026-03-31, onlineDate=1787294647945, onlineDateStr=2026-08-21, pubDate=1785772800000, pubDateStr=2026-08-04, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1787294647945, onlineIssueDateStr=2026-08-21, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1787294647945, creator=13701087609, updateTime=1787294647945, updator=13701087609, issue=Issue{id=1297570992835023717, tenantId=1146029695717560320, journalId=1192105938417971205, year='2026', volume='66', issue='8', pageStart='3681', pageEnd='4288', issueExtLink='null', onlineDate='null', pubDate='1785772800000', pubDateStr='2026-08-04', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1787294632395, creator='13701087609', updateTime=1787294931551, updator='13701087609', preIssue=null, nextIssue=null, articleTotal=null, ext={EN=IssueExt(id=1297572247670124783, tenantId=1146029695717560320, journalId=1192105938417971205, issueId=1297570992835023717, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1297572247670124784, tenantId=1146029695717560320, journalId=1192105938417971205, issueId=1297570992835023717, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null, downloadFileDto=null}, startPage=4030, endPage=4041, ext={EN=ArticleExt(id=1297571058261975936, articleId=1297571058056455039, tenantId=1146029695717560320, journalId=1192105938417971205, language=EN, title=Dissecting the ability of enterovirus A71 to traverse and retrogradely disrupt the blood-brain barrier and induce central nervous system inflammatory responses using an invitro blood-brain barrier-human cerebral organoid co-culture model, columnId=1192149543992045670, journalTitle=Acta Microbiologica Sinica, columnName=Research Article, runingTitle=null, highlight=null, articleAbstract=

[Objective] To investigate the ability of enterovirus A71 (EV-A71) to traverse the blood-brain barrier (BBB) and infect human cerebral organoids, as well as the impact of EV-A71 on BBB integrity following retrograde invasion into the central nervous system via neural routes, and to characterize the infection profiles of EV-A71 in the BBB and human cerebral organoids and the associated inflammatory responses. [Methods] An invitro BBB model was established by seeding human brain microvascular endothelial cells, brain vascular pericytes, and astrocytes into a Transwell system. Human cerebral organoids were generated from human induced pluripotent stem cells (hiPSCs), and a BBB-human cerebral organoid co-culture model was subsequently established. EV-A71 at three titers (10², 10⁴, and 10⁶ TCID₅₀) was used to infect each cell type of the BBB to evaluate viral replication kinetics and cytopathic effects (CPE) across different cell types. Subsequently, the BBB-human cerebral organoid co-culture model was infected with 10⁴ TCID₅₀ EV-A71 using two distinct routes: (1) an apical inoculation strategy, in which EV-A71 was directly applied to the human brain microvascular endothelial cell layer; and (2) a retrograde infection strategy, in which human cerebral organoids were first infected and subsequently co-cultured with the BBB model. Viral RNA copy numbers in culture supernatants and cells were quantified by RT-qPCR. BBB integrity was assessed by measuring transendothelial electrical resistance, and inflammatory responses were evaluated by determining the expression levels of inflammatory cytokines in human cerebral organoids using RT-qPCR. [Results] EV-A71 efficiently replicated in human brain microvascular endothelial cells, brain vascular pericytes, and astrocytes, inducing pronounced CPE. Among these cell types, astrocytes were the most susceptible to EV-A71 infection, showing the most rapid progression of cytopathic changes, followed by brain vascular pericytes. Human brain microvascular endothelial cells were relatively less susceptible, requiring higher viral titers to establish effective infection. Under apical inoculation conditions, EV-A71 replicated within the BBB, compromised BBB integrity, traversed the barrier to infect human cerebral organoids, and significantly activated inflammatory responses. In the retrograde infection model, EV-A71 replicated within human cerebral organoids, preferentially targeted neurons and astrocytes, triggered cellular inflammatory responses, and subsequently invaded the BBB in a retrograde manner, resulting in compromised structural integrity of the BBB. [Conclusion] EV-A71 can traverse the BBB to infect human cerebral organoids and activate inflammatory responses, and can also infect human cerebral organoids first and subsequently disrupt BBB integrity in a retrograde manner. This study systematically characterizes the infection dynamics of EV-A71 in an invitro BBB-human cerebral organoid model, provides a robust experimental model for in-depth analysis of the mechanisms underlying EV-A71 invasion of the central nervous system, and lays a research foundation for future investigations into viral pathogenesis and antiviral therapeutic development.

, authors=Juntong LIU, Weibang HUO, Roujian LU, Yao DENG, Jianfang ZHOU, Baoying HUANG, Wenjie TAN, authorsList=Juntong LIU, Weibang HUO, Roujian LU, Yao DENG, Jianfang ZHOU, Baoying HUANG, Wenjie TAN, authorCompany=null, correspAuthors=Wenjie TAN, authorNote=, correspAuthorsNote=, copyrightStatement=null, copyrightOwner=null, extLink=null, articleAbsUrl=null, sourceXml=null, magXml=null, pdfUrl=null, pdf=null, pdfFileSize=null, pdfExtLink=null, richHtmlUrl=null, mobilePdfUrl=null, reviewReport=null, pdfFirstPage=null, abstractGraph=null, abstractGraphContent=null, abstractVideo=null, citation=null, cebUrl=null, magXmlContent=null, mapNumber=null, fund=null), CN=ArticleExt(id=1297571059281191817, articleId=1297571058056455039, tenantId=1146029695717560320, journalId=1192105938417971205, language=CN, title=基于体外血脑屏障-人脑类器官共培养模型揭示肠道病毒A71型跨越与逆向破坏血脑屏障并诱导中枢炎症反应, columnId=1192149544164012138, journalTitle=微生物学报, columnName=研究报告, runingTitle=null, highlight=null, articleAbstract=

【目的】 探究肠道病毒A71型(enterovirus A71, EV-A71)突破血脑屏障(blood-brain barrier, BBB)感染人脑类器官的能力，以及EV-A71经神经途径逆行侵入中枢神经系统后对BBB完整性的影响，明确EV-A71在BBB及人脑类器官中的感染特征及其诱导的炎症反应。 【方法】 将人脑微血管内皮细胞、脑血管周细胞及星形胶质细胞接种于Transwell体系，构建体外BBB模型；使用人诱导多能干细胞诱导分化形成人脑类器官，建立BBB-人脑类器官共培养模型。以10²、10⁴、10⁶ TCID₅₀ 3种滴度EV-A71分别感染BBB各主要组成细胞，明确病毒在不同细胞类型中的复制动力学及细胞病变效应(cytopathic effect, CPE)。随后以10⁴ TCID₅₀ EV-A71通过2种方式感染BBB-人脑类器官共培养模型：(1)顶端接种法，直接感染人脑微血管内皮细胞侧；(2)反向感染法，先感染人脑类器官，再与BBB共培养。采用实时荧光定量逆转录PCR (real-time reverse transcription quantitative PCR, RT-qPCR)检测细胞培养上清及细胞内病毒核酸拷贝数，检测跨内皮电阻以评估BBB完整性，并以RT-qPCR检测人脑类器官中炎症因子的表达水平以评价炎症反应。 【结果】 EV-A71可在人脑微血管内皮细胞、脑血管周细胞及星形胶质细胞中复制并诱导明显CPE。其中星形胶质细胞对EV-A71最为敏感，病变进程最快；脑血管周细胞次之；人脑微血管内皮细胞相对不易感，需较高病毒滴度方可建立有效感染。顶端接种法结果显示，EV-A71可在BBB中复制，破坏BBB完整性，穿越屏障感染人脑类器官并显著激活炎症反应。反向感染法结果显示，EV-A71可在人脑类器官中复制，靶向感染神经元与星形胶质细胞，激活细胞炎症反应，并可进一步逆向侵袭BBB，导致其结构完整性受损。 【结论】 EV-A71可穿越血脑屏障感染人脑类器官并激活炎症反应，也可先感染人脑类器官后逆向破坏血脑屏障结构。本研究系统阐明了EV-A71对体外BBB及人脑类器官的感染特征，为深入解析EV-A71入侵中枢神经系统的感染及致病机制提供了有效的研究模型，并为后续致病机制的探索及抗病毒药物研发奠定了研究基础。

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Brain Research, 2020, 1730: 146647., articleTitle=Inflammatory stress induced by a combination of cytokines (IL-6, IL-17, TNF-α) leads to a loss of integrity on bEnd.3 endothelial cells in vitro BBB model, refAbstract=null)], funds=[Fund(id=1297571065136440258, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1297571058056455039, awardId=82061138008, language=EN, fundingSource=National Natural Science Foundation of China-International (Regional) Cooperation and Exchange Project(82061138008), fundOrder=null, country=null), Fund(id=1297571065203549123, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1297571058056455039, awardId=82061138008, language=CN, fundingSource=国家自然科学基金-国际(地区)合作与交流项目(82061138008), fundOrder=null, country=null)], companyList=[AuthorCompany(id=1297571059537044362, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1297571058056455039, xref=null, ext=[AuthorCompanyExt(id=1297571059545432971, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1297571058056455039, companyId=1297571059537044362, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=National Key Laboratory of Intelligent Tracking and Forecasting for Infectious Diseases, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing, China), AuthorCompanyExt(id=1297571059558015884, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1297571058056455039, companyId=1297571059537044362, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=中国疾病预防控制中心病毒病预防控制所，传染病溯源预警与智能决策全国重点实验室，国家卫生健康委员会生物安全重点实验室，北京)])], figs=[ArticleFig(id=1297571064440185786, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1297571058056455039, language=EN, label=Figure 1, caption=Replication kinetics and cytopathic effects of EV-A71 in the main cellular components of the BBB. A: Viral RNA copy numbers in culture supernatants of hBMECs infected with EV-A71 at titers of 10⁴ and 10⁶ TCID₅₀; B: Viral RNA copy numbers in culture supernatants of HBVPs infected with EV-A71 at titers of 10² and 10⁴ TCID₅₀; C: Viral RNA copy numbers in culture supernatants of astrocytes infected with EV-A71 at titers of 10² and 10⁴ TCID₅₀; D-F: CPE were observed and recorded every 24 h under an optical microscope., figureFileSmall=s5RFv0+u263B9WvuO9M22A==, figureFileBig=RrzRp9wXzk2e6JZLNHlaUw==, tableContent=null), ArticleFig(id=1297571064528266171, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1297571058056455039, language=CN, label=图1, caption=EV-A71在BBB的主要组成细胞中的复制动力学及细胞病变效应, figureFileSmall=s5RFv0+u263B9WvuO9M22A==, figureFileBig=RrzRp9wXzk2e6JZLNHlaUw==, tableContent=null), ArticleFig(id=1297571064708621244, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1297571058056455039, language=EN, label=Figure 2, caption=EV-A71 infects human cerebral organoids and disrupts BBB integrity. A: Schematic illustration of the EV-A71 infection procedure in the BBB-human cerebral organoid co-culture model; B: Viral RNA copy numbers in the apical and basolateral culture supernatants after EV-A71 infection of the co-culture model; C: TEER of the co-culture model after EV-A71 infection; D: Viral RNA copy numbers in human brain microvascular endothelial cells in the BBB model; E: Viral RNA copy numbers in brain pericytes and astrocytes in the BBB model; F: Viral RNA copy numbers in human cerebral organoids. ***: P<0.001., figureFileSmall=/hAdSAzNZgF9q4o6IaX41w==, figureFileBig=aG6uUCySqvaAYUxdN2hdhw==, tableContent=null), ArticleFig(id=1297571064784118717, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1297571058056455039, language=CN, label=图2, caption=EV-A71感染人脑类器官并破坏血脑屏障完整性, figureFileSmall=/hAdSAzNZgF9q4o6IaX41w==, figureFileBig=aG6uUCySqvaAYUxdN2hdhw==, tableContent=null), ArticleFig(id=1297571064851227582, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1297571058056455039, language=EN, label=Figure 3, caption=EV-A71 infection of neurons (A) and astrocytes (B) in human cerebral organoids., figureFileSmall=vXYYZLaVB4Lwy2UkeZuHxg==, figureFileBig=5FkymiunoxPDxi/dHLZBmQ==, tableContent=null), ArticleFig(id=1297571064922530751, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1297571058056455039, language=CN, label=图3, caption=EV-A71感染人脑类器官中神经元(A)与星形胶质细胞(B), figureFileSmall=vXYYZLaVB4Lwy2UkeZuHxg==, figureFileBig=5FkymiunoxPDxi/dHLZBmQ==, tableContent=null), ArticleFig(id=1297571064985445312, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1297571058056455039, language=EN, label=Figure 4, caption=EV-A71 infection of human cerebral organoids induces increased mRNA expression levels of the inflammatory cytokines IL-1β, IL-6, TNF-α, IL-8, IL-12, and MCP-1. **: P<0.01., figureFileSmall=ulrKqBqRFClnQeKYGVOAoQ==, figureFileBig=60OgUZ1vF1w7BurEBoZkRw==, 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