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

基本信息

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
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/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

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.

抽象的蝙蝠作为多种 RNA 病毒的天然储存库发挥着重要作用，有可能造成严重的后果。在本提案中，我们重点关注 Lloviu 病毒 (LLOV)，这是一种尚未得到充分研究的丝状病毒。尽管 LLOV 对欧洲的施赖伯氏蝙蝠（Miniopterus schreibersii）具有致病潜力。尚不清楚，LLOV 与高致病性埃博拉病毒和马尔堡病毒的密切关系引起关注潜在的溢出事件可能导致人类爆发疫情。 LLOV 于 2002 年在西班牙首次检测到 Schreiber 的球棒，然后于 2016 年在匈牙利再次检测到。西班牙和匈牙利 LLOV RNA 基因组的比较表明，细胞的 RNA 编辑脱氨酶，例如 ADAR 和 APOBEC，可能在 LLOV 序列多样化中发挥作用。我们建议探索宿主介导的 RNA 编辑是否驱动 LLOV 序列的分歧和进化施赖伯的蝙蝠。在目标 1 中，我们建议从地理不同的群体中对施赖伯氏蝙蝠进行采样，并获得 LLOV 序列我们将进一步开发基于高度敏感的工具来进行比较分析。液滴 RT-PCR 和 RNA FISH 可以确定 ADAR 和 APOBEC 的表达模式 LLOV 感染的蝙蝠细胞培养物和受感染动物的血液样本。在目标 2 中，根据确定的 ADAR 和 APOBEC 表达模式，我们将敲除选定的 ADAR 和/或可能参与蝙蝠细胞系中 LLOV RNA 编辑的 APOBEC 基因，并检查这些基因在连续传递实验和细胞培养中的 LLOV 序列多样化和病毒适应性感染研究。完成这项工作后，我们将揭示宿主特异性 RNA 编辑器是否是 LLOV 的驱动因素这项工作将有助于我们理解宿主驱动的病毒序列分歧。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

期刊论文数量（2）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

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.