Reverse genetics systems for contemporary isolates of respiratory syncytial virus enable rapid evaluation of antibody escape mutants.
cell-to-cell fusion
escape mutants
respiratory syncytial virus
reverse genetics
Journal
Proceedings of the National Academy of Sciences of the United States of America
ISSN: 1091-6490
Titre abrégé: Proc Natl Acad Sci U S A
Pays: United States
ID NLM: 7505876
Informations de publication
Date de publication:
06 04 2021
06 04 2021
Historique:
entrez:
3
4
2021
pubmed:
4
4
2021
medline:
20
11
2021
Statut:
ppublish
Résumé
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 laboratory-adapted 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.
Identifiants
pubmed: 33811145
pii: 2026558118
doi: 10.1073/pnas.2026558118
pmc: PMC8040649
pii:
doi:
Substances chimiques
Antiviral Agents
0
Palivizumab
DQ448MW7KS
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Déclaration de conflit d'intérêts
Competing interest statement: A patent application (PCT/EP2019/054775) entitled “Improved Pneumovirus reverse genetics” was filed on 26 February 2019. However, this application was discontinued and allowed to lapse in August 2020.
Références
Virology. 1995 Apr 20;208(2):478-84
pubmed: 7747420
J Virol. 2000 Jul;74(14):6643-7
pubmed: 10864679
Nat Rev Microbiol. 2019 Apr;17(4):233-245
pubmed: 30723301
Mol Biol Evol. 2015 Jan;32(1):268-74
pubmed: 25371430
J Gen Virol. 2003 Nov;84(Pt 11):3115-3120
pubmed: 14573817
Curr Infect Dis Rep. 2020 Jan 24;22(1):3
pubmed: 31980966
Proc Natl Acad Sci U S A. 1986 Oct;83(19):7462-6
pubmed: 3532115
Pediatr Res. 2015 Oct;78(4):380-8
pubmed: 26107392
Pediatr Res. 2004 Dec;56(6):914-7
pubmed: 15470202
J Gen Virol. 2015 Sep;96(9):2543-2556
pubmed: 26297201
J Virol. 2016 Nov 14;90(23):10612-10628
pubmed: 27654298
Br J Exp Pathol. 1964 Dec;45:647-55
pubmed: 14245166
J Virol. 2011 Aug;85(15):7788-96
pubmed: 21613394
Biomed Res Int. 2018 Jan 15;2018:8431243
pubmed: 29568767
J Virol. 2005 May;79(9):5326-36
pubmed: 15827147
J Virol. 1998 May;72(5):4467-71
pubmed: 9557743
Arch Virol. 1994;136(1-2):133-47
pubmed: 8002781
Am J Hyg. 1957 Nov;66(3):281-90
pubmed: 13478578
J Gen Virol. 2019 Jul;100(7):1112-1122
pubmed: 31184573
J Gen Virol. 1997 Aug;78 ( Pt 8):1885-9
pubmed: 9266983
mBio. 2016 Mar 15;7(2):e00235
pubmed: 26980833
EMBO J. 1994 Sep 15;13(18):4195-203
pubmed: 7925265
PLoS Pathog. 2015 Dec 11;11(12):e1005318
pubmed: 26658574
PLoS One. 2015 Jun 24;10(6):e0130517
pubmed: 26107373
Virol J. 2014 Aug 08;11:142
pubmed: 25107552
J Gen Virol. 2020 Oct;101(10):1037-1046
pubmed: 32692644
Pathogens. 2019 May 21;8(2):
pubmed: 31117229
Virus Genes. 2010 Apr;40(2):212-21
pubmed: 20111897
Proc Natl Acad Sci U S A. 2000 May 9;97(10):5516-21
pubmed: 10805807
J Infect Dis. 2011 Mar 1;203(5):674-82
pubmed: 21208913
J Infect Dis. 1997 Nov;176(5):1215-24
pubmed: 9359721
Proc Natl Acad Sci U S A. 1995 Dec 5;92(25):11563-7
pubmed: 8524804
J Virol. 2016 Jul 27;90(16):7508-7518
pubmed: 27279612
Thorax. 2006 Jul;61(7):611-5
pubmed: 16537670
J Gen Virol. 1985 Oct;66 ( Pt 10):2111-24
pubmed: 2413163
J Virol. 2015 Sep;89(18):9499-510
pubmed: 26157122
J Virol. 2015 Mar;89(5):2849-56
pubmed: 25540371
Virology. 2012 Dec 5;434(1):129-36
pubmed: 23062737
Front Microbiol. 2019 May 03;10:873
pubmed: 31130923
Fukushima J Med Sci. 2017 Dec 19;63(3):127-134
pubmed: 28867684
Lancet. 2019 Aug 31;394(10200):757-779
pubmed: 31257127
Proc Soc Exp Biol Med. 1956 Jul;92(3):544-9
pubmed: 13359460
Adv Virus Res. 1999;54:423-51
pubmed: 10547682
J Virol. 2010 May;84(10):5303-13
pubmed: 20219902
Viruses. 2019 Nov 06;11(11):
pubmed: 31698728
Arch Virol. 2020 May;165(5):1069-1077
pubmed: 32144544
J Med Virol. 2007 Dec;79(12):1951-8
pubmed: 17935185
Am J Epidemiol. 1966 Mar;83(2):299-313
pubmed: 5933417
PLoS One. 2017 Apr 17;12(4):e0175792
pubmed: 28414749
J Virol. 2000 May;74(10):4831-8
pubmed: 10775622
J Virol. 2011 Jun;85(12):5782-93
pubmed: 21471228
Nat Commun. 2014 Oct 03;5:5104
pubmed: 25277263
J Gen Virol. 2020 Oct;101(10):1056-1068
pubmed: 32723429
J Infect Dis. 2019 Jan 29;219(4):514-516
pubmed: 30215722
Int J Mol Sci. 2017 Aug 06;18(8):
pubmed: 28783078
Virology. 1998 Nov 10;251(1):206-14
pubmed: 9813216
J Infect Dis. 1985 Apr;151(4):626-33
pubmed: 2579169
Emerg Microbes Infect. 2018 Dec 5;7(1):201
pubmed: 30514855
J Infect Dis. 2020 Oct 1;222(9):1417-1421
pubmed: 31974551
Virology. 1994 May 1;200(2):801-5
pubmed: 8178462
Proc Natl Acad Sci U S A. 2012 Mar 27;109(13):5040-5
pubmed: 22411804
J Virol. 2003 Jun;77(11):6450-65
pubmed: 12743302
Brief Bioinform. 2019 Jul 19;20(4):1160-1166
pubmed: 28968734
Nature. 2020 Jun;582(7813):561-565
pubmed: 32365353
J Virol. 2015 Aug;89(16):8258-66
pubmed: 26018171
Virol J. 2007 Jun 05;4:51
pubmed: 17550613
J Virol. 1983 Jul;47(1):171-7
pubmed: 6345804
Proc Natl Acad Sci U S A. 1997 Dec 9;94(25):13961-6
pubmed: 9391135
Methods Mol Biol. 2016;1442:13-32
pubmed: 27464684
PLoS One. 2012;7(3):e32807
pubmed: 22470426
J Gen Virol. 2013 Dec;94(Pt 12):2627-2635
pubmed: 24092758
Clin Microbiol Rev. 2000 Jan;13(1):1-15, table of contents
pubmed: 10627488
Nat Methods. 2017 Jun;14(6):587-589
pubmed: 28481363
J Gen Virol. 2020 Nov;101(11):1156-1169
pubmed: 32821033
J Gen Virol. 1992 Sep;73 ( Pt 9):2225-34
pubmed: 1383404
Clin Infect Dis. 2010 Jul 15;51(2):185-8
pubmed: 20528472