Potent Single-domain Antibodies That Arrest Respiratory Syncytial Virus Fusion Protein in Its Prefusion State

Overview

Journal Nat Commun

Specialty Biology

Date 2017 Feb 15

PMID 28194013

Citations 48

Authors

Iebe Rossey

Morgan S A Gilman

Stephanie C Kabeche

Koen Sedeyn

Daniel Wrapp

Masaru Kanekiyo

Man Chen

Vicente Mas

Jan Spitaels

Jose A Melero

Barney S Graham

Bert Schepens

Jason S McLellan

Xavier Saelens

Affiliations

Soon will be listed here.

Abstract

Human respiratory syncytial virus (RSV) is the main cause of lower respiratory tract infections in young children. The RSV fusion protein (F) is highly conserved and is the only viral membrane protein that is essential for infection. The prefusion conformation of RSV F is considered the most relevant target for antiviral strategies because it is the fusion-competent form of the protein and the primary target of neutralizing activity present in human serum. Here, we describe two llama-derived single-domain antibodies (VHHs) that have potent RSV-neutralizing activity and bind selectively to prefusion RSV F with picomolar affinity. Crystal structures of these VHHs in complex with prefusion F show that they recognize a conserved cavity formed by two F protomers. In addition, the VHHs prevent RSV replication and lung infiltration of inflammatory monocytes and T cells in RSV-challenged mice. These prefusion F-specific VHHs represent promising antiviral agents against RSV.

Citing Articles

DS2 designer pre-fusion F vaccine induces strong and protective antibody response against RSV infection.

Yang Y, Wang R, Guo F, Zhao T, Lei Y, Yang Q NPJ Vaccines. 2024; 9(1):258.

PMID: 39741146 PMC: 11688450. DOI: 10.1038/s41541-024-01059-9.

Rational design of uncleaved prefusion-closed trimer vaccines for human respiratory syncytial virus and metapneumovirus.

Lee Y, Han J, Zhang Y, Ward G, Gomes K, Auclair S Nat Commun. 2024; 15(1):9939.

PMID: 39550381 PMC: 11569192. DOI: 10.1038/s41467-024-54287-x.

Direct and Ultrasensitive Bioluminescent Detection of Intact Respiratory Viruses.

Grawe A, van der Veer H, Jongkees S, Flipse J, Rossey I, de Vries R ACS Sens. 2024; 9(10):5550-5560.

PMID: 39375866 PMC: 11519905. DOI: 10.1021/acssensors.4c01855.

Structural basis for potent neutralization of human respirovirus type 3 by protective single-domain camelid antibodies.

Johnson N, van Scherpenzeel R, Bakkers M, Ramamohan A, van Overveld D, Le L Nat Commun. 2024; 15(1):5458.

PMID: 38937429 PMC: 11211449. DOI: 10.1038/s41467-024-49757-1.

Respiratory Syncytial Virus in Adult Patients at a Tertiary Care Hospital in Germany: Clinical Features and Molecular Epidemiology of the Fusion Protein in the Severe Respiratory Season of 2022/2023.

Honemann M, Maier M, Frille A, Thiem S, Bergs S, Williams T Viruses. 2024; 16(6).

PMID: 38932235 PMC: 11209376. DOI: 10.3390/v16060943.

References

Hotard A, Shaikh F, Lee S, Yan D, Teng M, Plemper R . A stabilized respiratory syncytial virus reverse genetics system amenable to recombination-mediated mutagenesis. Virology. 2012; 434(1):129-36. PMC: 3492879. DOI: 10.1016/j.virol.2012.09.022. View

Battye T, Kontogiannis L, Johnson O, Powell H, Leslie A . iMOSFLM: a new graphical interface for diffraction-image processing with MOSFLM. Acta Crystallogr D Biol Crystallogr. 2011; 67(Pt 4):271-81. PMC: 3069742. DOI: 10.1107/S0907444910048675. View

Hart T, Cook R, Zia-Amirhosseini P, Minthorn E, Sellers T, Maleeff B . Preclinical efficacy and safety of mepolizumab (SB-240563), a humanized monoclonal antibody to IL-5, in cynomolgus monkeys. J Allergy Clin Immunol. 2001; 108(2):250-7. DOI: 10.1067/mai.2001.116576. View

Magro M, Mas V, Chappell K, Vazquez M, Cano O, Luque D . Neutralizing antibodies against the preactive form of respiratory syncytial virus fusion protein offer unique possibilities for clinical intervention. Proc Natl Acad Sci U S A. 2012; 109(8):3089-94. PMC: 3286924. DOI: 10.1073/pnas.1115941109. View

Corti D, Bianchi S, Vanzetta F, Minola A, Perez L, Agatic G . Cross-neutralization of four paramyxoviruses by a human monoclonal antibody. Nature. 2013; 501(7467):439-43. DOI: 10.1038/nature12442. View

Schoonooghe S, Kaigorodov V, Zawisza M, Dumolyn C, Haustraete J, Grooten J . Efficient production of human bivalent and trivalent anti-MUC1 Fab-scFv antibodies in Pichia pastoris. BMC Biotechnol. 2009; 9:70. PMC: 2736937. DOI: 10.1186/1472-6750-9-70. View

Evans P, Murshudov G . How good are my data and what is the resolution?. Acta Crystallogr D Biol Crystallogr. 2013; 69(Pt 7):1204-14. PMC: 3689523. DOI: 10.1107/S0907444913000061. View

Van Audenhove I, Gettemans J . Nanobodies as Versatile Tools to Understand, Diagnose, Visualize and Treat Cancer. EBioMedicine. 2016; 8:40-48. PMC: 4919472. DOI: 10.1016/j.ebiom.2016.04.028. View

McCoy A, Grosse-Kunstleve R, Adams P, Winn M, Storoni L, Read R . Phaser crystallographic software. J Appl Crystallogr. 2009; 40(Pt 4):658-674. PMC: 2483472. DOI: 10.1107/S0021889807021206. View

10.

Behera A, Matsuse H, Kumar M, Kong X, Lockey R, Mohapatra S . Blocking intercellular adhesion molecule-1 on human epithelial cells decreases respiratory syncytial virus infection. Biochem Biophys Res Commun. 2001; 280(1):188-95. DOI: 10.1006/bbrc.2000.4093. View

11.

Garcia O, Martin M, Dopazo J, Arbiza J, Frabasile S, Russi J . Evolutionary pattern of human respiratory syncytial virus (subgroup A): cocirculating lineages and correlation of genetic and antigenic changes in the G glycoprotein. J Virol. 1994; 68(9):5448-59. PMC: 236945. DOI: 10.1128/JVI.68.9.5448-5459.1994. View

12.

Schepens B, Ibanez L, De Baets S, Hultberg A, Bogaert P, De Bleser P . Nanobodies® specific for respiratory syncytial virus fusion protein protect against infection by inhibition of fusion. J Infect Dis. 2011; 204(11):1692-701. DOI: 10.1093/infdis/jir622. View

13.

Teng M, Whitehead S, Collins P . Contribution of the respiratory syncytial virus G glycoprotein and its secreted and membrane-bound forms to virus replication in vitro and in vivo. Virology. 2001; 289(2):283-96. DOI: 10.1006/viro.2001.1138. View

14.

Kwakkenbos M, Diehl S, Yasuda E, Bakker A, van Geelen C, Lukens M . Generation of stable monoclonal antibody-producing B cell receptor-positive human memory B cells by genetic programming. Nat Med. 2009; 16(1):123-8. PMC: 2861345. DOI: 10.1038/nm.2071. View

15.

Wu S, Schmidt A, Beil E, Day N, Branigan P, Liu C . Characterization of the epitope for anti-human respiratory syncytial virus F protein monoclonal antibody 101F using synthetic peptides and genetic approaches. J Gen Virol. 2007; 88(Pt 10):2719-2723. DOI: 10.1099/vir.0.82753-0. View

16.

van Heeke G, Allosery K, De Brabandere V, De Smedt T, Detalle L, De Fougerolles A . Nanobodies® as inhaled biotherapeutics for lung diseases. Pharmacol Ther. 2016; 169:47-56. DOI: 10.1016/j.pharmthera.2016.06.012. View

17.

Glezen W, Taber L, Frank A, KASEL J . Risk of primary infection and reinfection with respiratory syncytial virus. Am J Dis Child. 1986; 140(6):543-6. DOI: 10.1001/archpedi.1986.02140200053026. View

18.

Lin-Cereghino J, Wong W, Xiong S, Giang W, Luong L, Vu J . Condensed protocol for competent cell preparation and transformation of the methylotrophic yeast Pichia pastoris. Biotechniques. 2005; 38(1):44, 46, 48. PMC: 2504082. DOI: 10.2144/05381BM04. View

19.

Wu H, Pfarr D, Johnson S, Brewah Y, Woods R, Patel N . Development of motavizumab, an ultra-potent antibody for the prevention of respiratory syncytial virus infection in the upper and lower respiratory tract. J Mol Biol. 2007; 368(3):652-65. DOI: 10.1016/j.jmb.2007.02.024. View

20.

Johnson S, Oliver C, Prince G, Hemming V, Pfarr D, Wang S . Development of a humanized monoclonal antibody (MEDI-493) with potent in vitro and in vivo activity against respiratory syncytial virus. J Infect Dis. 1997; 176(5):1215-24. DOI: 10.1086/514115. View