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

阐明施赖伯氏蝙蝠对 Lloviu 病毒感染的体外和体内免疫反应

• 批准号: 10579294
• 负责人: Elke C Muhlberger
• 金额: $ 18.63万
• 依托单位: BOSTON UNIVERSITY MEDICAL CAMPUS
• 依托单位国家: 美国
• 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/10579294
• 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; Marburgvirus; Mediating; Pathogenicity; Pattern; Play; Population; Procedures; Process; Public Health; RNA; RNA Editing; RNA Viruses; Recombinants; Research; Research Personnel; Reverse Transcriptase Polymerase Chain Reaction; Risk Assessment; Role; Rousettus; Sampling; Serial Passage; Southern Europe; Spain; System; Viral; Viral Genome; Virus; Virus Diseases; Visualization; Work; Zoonoses; apolipoprotein B mRNA editing enzyme; bat-borne; comparative; deep sequencing; dsRNA adenosine deaminase; experimental study; fitness; genome editing; in vivo; insight; member; novel; pandemic disease; pathogenic virus; polypeptide; reverse genetics; spillover event; tool; viral RNA; viral fitness

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.

摘要 蝙蝠是许多核糖核酸病毒的天然储存库，有可能引起重大的 对人类的伤害。在这项建议中，我们将重点放在Lloviu病毒(LLOV)上，这是一种正在传播的未被研究的丝状病毒 在欧洲的Schreiber蝙蝠(Miniopterus Schreibersii)。尽管LLOV对人类的致病潜力 LLOV与高致病性埃博拉和马尔堡病毒的密切关系引起了人们的关注 潜在的溢出事件可能导致人类之间的疫情爆发。 LLOV于2002年在西班牙首次发现Schreiber的蝙蝠，然后于2016年在匈牙利再次发现。数列 对西班牙和匈牙利LLOV RNA基因组的比较表明，通过细胞编辑RNA 脱氨酶，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编辑是否驱动LLOV序列的分化和进化 施赖伯的蝙蝠。 在目标1中，我们建议从地理上不同的群体中采样Schreiber蝙蝠并获得LLOV序列 来自受感染蝙蝠的信息，用于比较分析。我们将进一步开发基于高度敏感的工具 滴状RT-PCR和RNA FISH检测ADARs和APOBECs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中。 在目标2中，基于确定的ADAR和APOBEC表达模式，我们将敲除选择ADAR 和/或可能参与LLOV RNA编辑的APOBEC基因在BAT细胞系中的作用 这些基因在LLOV序列多样性和连续传代实验和细胞培养中的病毒适合性 感染研究。 在完成这项工作后，我们将揭示特定于宿主的RNA编辑是否是LLOV的驱动因素 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从蝙蝠到人类的潜在LLOV溢出事件的风险 病毒序列的变化。

项目成果

Isolation and genome characterization of Lloviu virus from Italian Schreibers's bats.

• DOI: 10.1038/s41598-023-38364-7
• 发表时间: 2023-07-13
• 期刊: SCIENTIFIC REPORTS
• 影响因子: 4.6
• 作者: Toth, Gabor E.;Hume, Adam J.;Suder, Ellen L.;Zeghbib, Safia;abraham, Agota;Lanszki, Zsofia;Varga, Zsaklin;Tauber, Zsofia;Foeldes, Fanni;Zana, Brigitta;Scaravelli, Dino;Scicluna, Maria Teresa;Pereswiet-Soltan, Andrea;Goerfoel, Tamas;Terregino, Calogero;De Benedictis, Paola;Garcia-Dorival, Isabel;Alonso, Covadonga;Jakab, Ferenc;Muehlberger, Elke;Leopardi, Stefania;Kemenesi, Gabor
• 通讯作者: Kemenesi, Gabor

Labeling Ebola Virus with a Self-Splicing Fluorescent Reporter.

• DOI: 10.3390/microorganisms10112110
• 发表时间: 2022-10-26
• 期刊: Microorganisms
• 影响因子: 4.5