气候变化领域集成服务门户(CCPortal): Reverse genetics systems for contemporary isolates of respiratory syncytial virus enable rapid evaluation of antibody escape mutants

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DOI	10.1073/pnas.2026558118
	Reverse genetics systems for contemporary isolates of respiratory syncytial virus enable rapid evaluation of antibody escape mutants
	Jo W.K.; Schadenhofer A.; Habierski A.; Kaiser F.K.; Saletti G.; Ganzenmueller T.; Hage E.; Haid S.; Pietschmann T.; Hansen G.; Schulz T.F.; Rimmelzwaan G.F.; Osterhaus A.D.M.E.; Ludlow M.
发表日期	2021
ISSN	00278424
卷号	118 期号:14
英文摘要	Human respiratory syncytial virus (RSV) is the leading cause of acute lower respiratory infection in children under 5 y of age. In the absence of a safe and effective vaccine and with limited options for therapeutic interventions, uncontrolled epidemics of RSV occur annually worldwide. Existing RSV reverse genetics systems have been predominantly based on older laboratoryadapted strains such as A2 or Long. These strains are not representative of currently circulating genotypes and have a convoluted passage history, complicating their use in studies on molecular determinants of viral pathogenesis and intervention strategies. In this study, we have generated reverse genetics systems for clinical isolates of RSV-A (ON1, 0594 strain) and RSV-B (BA9, 9671 strain) in which the full-length complementary DNA (cDNA) copy of the viral antigenome is cloned into a bacterial artificial chromosome (BAC). Additional recombinant (r) RSVs were rescued expressing enhanced green fluorescent protein (EGFP), mScarlet, or NanoLuc luciferase from an additional transcription unit inserted between the P and M genes. Mutations in antigenic site II of the F protein conferring escape from palivizumab neutralization (K272E, K272Q, S275L) were investigated using quantitative cell-fusion assays and rRSVs via the use of BAC recombineering protocols. These mutations enabled RSV-A and -B to escape palivizumab neutralization but had differential impacts on cell-to-cell fusion, as the S275L mutation resulted in an almost-complete ablation of syncytium formation. These reverse genetics systems will facilitate future cross-validation efficacy studies of novel RSV therapeutic intervention strategies and investigations into viral and host factors necessary for virus entry and cell-to-cell spread. © 2021 National Academy of Sciences. All rights reserved.
英文关键词	Cell-to-cell fusion; Escape mutants; Respiratory syncytial virus; Reverse genetics
语种	英语
来源期刊	Proceedings of the National Academy of Sciences of the United States of America
文献类型	期刊论文
条目标识符	http://gcip.llas.ac.cn/handle/2XKMVOVA/179950
作者单位	Research Center for Emerging Infections and Zoonoses, University of Veterinary Medicine Hannover, Hannover, 30559, Germany; Institute of Virology, Hannover Medical School, Hannover, 30625, Germany; Institute for Experimental Virology, Twincore Centre for Experimental and Clinical Infection Research, a joint venture of Hannover Medical School and Helmholtz Centre for Infection Research, Hannover, 30625, Germany; Cluster of Excellence RESIST (EXC 2155), Hannover Medical School, Hannover, 30625, Germany; Department for Pediatric Pneumology Allergy and Neonatology, Hannover Medical School, Hannover, 30625, Germany
推荐引用方式 GB/T 7714	Jo W.K.,Schadenhofer A.,Habierski A.,et al. Reverse genetics systems for contemporary isolates of respiratory syncytial virus enable rapid evaluation of antibody escape mutants[J],2021,118(14).
APA	Jo W.K..,Schadenhofer A..,Habierski A..,Kaiser F.K..,Saletti G..,...&Ludlow M..(2021).Reverse genetics systems for contemporary isolates of respiratory syncytial virus enable rapid evaluation of antibody escape mutants.Proceedings of the National Academy of Sciences of the United States of America,118(14).
MLA	Jo W.K.,et al."Reverse genetics systems for contemporary isolates of respiratory syncytial virus enable rapid evaluation of antibody escape mutants".Proceedings of the National Academy of Sciences of the United States of America 118.14(2021).