ROLE OF RHESUS ROTAVIRUS GENE 4 IN BILIARY ATRESIA

恒河猴轮状病毒基因 4 在胆道闭锁中的作用

• 批准号: 8358153
• 负责人: KAROL SESTAK
• 金额: $ 3.72万
• 依托单位: TULANE UNIVERSITY OF LOUISIANA
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-05-01 至 2012-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8358153
• 关键词: Affect; Attenuated; Biliary; Biliary Atresia; Binding; Biological Assay; Childhood; Disease; Double Stranded RNA Virus; Epithelial Cells; Etiology; Funding; Genes; Grant; In Vitro; Infection; Macaca mulatta; Modeling; Mus; NR4A1 gene; National Center for Research Resources; Obstruction; Pathogenesis; Perinatal Infection; Phenotype; Play; Primates; Principal Investigator; Property; Proteins; Research; Research Infrastructure; Resources; Role; Rotavirus; Source; Translating; Tropism; United States National Institutes of Health; Viral; base; cholangiocyte; cost; gain of function; in vivo; loss of function; mortality

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Biliary atresia (BA) is a devastating disease of childhood. The etiology of BA remains uncertain; however, increasing evidence supports a viral component in disease pathogenesis. The murine model of BA is induced by perinatal infection with rhesus rotavirus (RRV) but not by other strains of rotavirus an example of which is TUCH. Rotavirus is a double-stranded RNA virus that contains 11 gene segments. To determine which RRV gene segment is responsible for disease pathogenesis, we used the parental strains RRV and TUCH and the rotavirus property of reassortment to generate a complete set of single gene reassortants. Eleven single-gene "loss-of-function" reassortants in which a TUCH gene replaced its RRV equivalent and eleven reciprocal single gene "gain-of-function" reassortants in which a RRV gene replaced its TUCH equivalent were generated. Infection of mice with the "loss-of-function" reassortant RT4 where gene segment 4 from TUCH was placed on an RRV background eliminated RRV's ability to cause murine BA. In a reciprocal fashion, the "gain-of-function" reassortant TR4 resulted in murine BA with a mortality rate of 95%. Cholangiocyte binding and infectivity assays revealed RT4 binding ratio and titer (both in vivo and in vitro) were significantly attenuated as compared with RRV; moreover, TR4 binding and titer were significantly higher than TUCH. Reassortant RT3 and TR3 induced an intermediate phenotype. Gene segment 4 through its translated gene product VP4 plays a significant role governing RRV tropism for the biliary epithelial cell and induces the murine model of biliary atresia. The basis for tropism and how it induces biliary obstruction requires further study. Substitution of RRV gene segment 3 did not affect viral infectivity in vitro, but altered the in vivo effect.

这个子项目是许多利用资源的研究子项目之一 由NIH/NCRR资助的中心拨款提供。子项目的主要支持 而子项目的主要调查员可能是由其他来源提供的， 包括其它NIH来源。 列出的子项目总成本可能 代表子项目使用的中心基础设施的估计数量， 而不是由NCRR赠款提供给子项目或子项目工作人员的直接资金。 胆道闭锁（BA）是一种毁灭性的儿童疾病。BA的病因仍然不确定，然而，越来越多的证据支持疾病发病机制中的病毒成分。BA的鼠模型是由恒河猴轮状病毒（RRV）的围产期感染诱导的，但不是由其他轮状病毒株（TUCH）诱导的。轮状病毒是一种双链RNA病毒，含有11个基因片段。为了确定哪一个RRV基因片段负责疾病的发病机制，我们使用亲本株RRV和TUCH以及轮状病毒的重配特性来产生一套完整的单基因重组体。产生了11个TUCH基因取代其RRV等价物的单基因“功能丧失”的resistant和11个RRV基因取代其TUCH等价物的相互单基因“功能获得”的resistant。用“功能丧失”的重组体RT 4感染小鼠，其中来自TUCH的基因片段4被置于RRV背景上，消除了RRV引起小鼠BA的能力。以相反的方式，“功能获得性”抑制剂TR 4导致小鼠BA，死亡率为95%。胆管细胞结合和感染性测定显示，RT 4结合率和滴度（体内和体外）与RRV相比显著减弱;此外，TR 4结合率和滴度显著高于TUCH。抗性株RT 3和TR 3诱导中间型。 基因片段4通过其翻译的基因产物VP 4在控制RRV对胆管上皮细胞的嗜性并诱导胆道闭锁的小鼠模型中起重要作用。 归经的基础及其如何诱发胆道梗阻尚需进一步研究。RRV基因片段3的取代不影响病毒的体外感染性，但改变了体内效果。