Elucidating the immune response of Schreiber's bats to Lloviu virus infection in vitro and in vivo

阐明施赖伯蝙蝠对 Lloviu 病毒感染的体外和体内免疫反应

基本信息

批准号：
10458900
负责人：
Elke C Muhlberger
金额：
$ 24.38万
依托单位：
BOSTON UNIVERSITY MEDICAL CAMPUS
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-02-24 至 2024-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10458900
关键词：
Animals Benign Biological Assay Blood Blood specimen Cell Culture Techniques Cell Line Cells Chiroptera Complex Country Data Deaminase Disease Disease Outbreaks Eastern Europe Ebola virus Enzymes Epidemic Europe Evolution Family Fatality rate Filovirus Fruit Gene Expression Genes Genetic Drift Genome Geography Goals Human Hungary Immune response In Vitro Infection Knock-in Knock-out Lead Light Marburgvirus Mediating Pathogenicity Pattern Play Population Procedures Process Public Health RNA RNA Editing RNA Viruses Recombinants Research Reverse Transcriptase Polymerase Chain Reaction Risk Role Rousettus Sampling Serial Passage Southern Europe Spain System Viral Viral Genome Virus Virus Diseases Work Zoonoses adenosine deaminase apolipoprotein B mRNA editing enzyme base bat-borne comparative deep sequencing experimental study fitness genome editing in vivo insight member novel pandemic disease pathogenic virus polypeptide reverse genetics spillover event tool viral RNA viral fitness

项目摘要

ABSTRACT Bats play an important role as natural reservoirs of numerous RNA viruses with the potential to cause significant harm to humans. In this proposal, we focus on Lloviu virus (LLOV), an under-investigated filovirus that circulates in Schreiber’s bats (Miniopterus schreibersii) in Europe. Although the pathogenic potential of LLOV for humans is not known, the close relationship of LLOV to the highly pathogenic Ebola and Marburg viruses raises concerns that a potential spillover event could lead to an outbreak among humans. LLOV was first detected Schreiber’s bats in Spain in 2002 and then again in Hungary in 2016. Sequence comparison of the Spanish and the Hungarian LLOV RNA genomes suggests that RNA editing by cellular deaminases, such as ADAR and APOBEC, might play a role in LLOV sequence diversification. In this application, we propose to explore if host-mediated RNA editing drives LLOV sequence divergence and evolution in Schreiber’s bats. In Aim 1, we propose to sample Schreiber’s bats from geographically distinct colonies and obtain LLOV sequence information from infected bats for comparative analysis. We will further develop tools based on highly sensitive droplet RT-PCR and RNA FISH that allow to determine the expression pattern of ADARs and APOBECs in LLOV-infected bat cell culture and in blood samples from infected animals. In Aim 2, based on the determined ADAR and APOBEC expression patterns, we will knockout select ADAR and/or APOBEC genes that might be involved in LLOV RNA editing in the bat cell line and examine the role of these genes in LLOV sequence diversification and viral fitness in serial passaging experiments and cell culture infection studies. Upon completion of this work, we will have revealed whether host-specific RNA editors are the drivers of LLOV evolution in bat cells. This work will contribute to our understanding of host-driven viral sequence divergence and might help assess the risk of potential LLOV spillover events from bats to humans through host-driven changes in viral sequences.

抽象的蝙蝠是众多RNA病毒的自然储层起重要作用，有可能引起重要的对人类的伤害。在此提案中，我们专注于LLOVIU病毒（LLOV），这是一种循环的未经评价的污物病毒在欧洲的Schreiber的蝙蝠（Miniopterus Schreibersii）中。虽然LLOV对人的致病潜力尚不清楚，LLOV与高致病性埃博拉病毒的密切关系引起了人们的关注潜在的Spilover事件可能导致人类爆发。 LLOV于2002年首次在西班牙被发现在西班牙，然后在2016年再次在匈牙利。西班牙语和匈牙利LLOV RNA基因组的比较表明通过细胞编辑RNA 脱氨酶，例如ADAR和APOBEC，可能在LLOV序列多样化中发挥作用。在此应用程序中我们建议探索宿主介导的RNA编辑是否会驱动LLOV序列发散和进化 Schreiber的蝙蝠。在AIM 1中，我们建议从地理上不同的菌落中采样Schreiber的蝙蝠并获得LLOV序列来自受感染的蝙蝠的信息进行比较分析。我们将进一步开发基于高度敏感的工具液滴RT-PCR和RNA鱼，允许确定ADAR和APOBEC的表达模式 LLOV感染的蝙蝠细胞培养和感染动物的血液样本。在AIM 2中，基于确定的ADAR和APOBEC表达模式，我们将淘汰选择ADAR 和/或APOBEC基因可能参与蝙蝠细胞系中LLOV RNA编辑并检查的作用在LLOV序列多样化和串行传递实验中的病毒适应性和细胞培养中的这些基因感染研究。完成这项工作后，我们将透露主机特定的RNA编辑是否是LLOV的驱动力蝙蝠细胞的进化。这项工作将有助于我们理解宿主驱动的病毒序列差异并可能有助于评估通过宿主驱动的潜在LLOV Spilover事件从蝙蝠到人的风险病毒序列的变化。