Respiratory syncytial virus entry and how to block it

Abstract

Respiratory syncytial virus (RSV) is a leading cause of lower respiratory tract disease in young children and elderly people. Although the virus was isolated in 1955, an effective RSV vaccine has not been developed, and the only licensed intervention is passive immunoprophylaxis of high-risk infants with a humanized monoclonal antibody. During the past 5 years, however, there has been substantial progress in our understanding of the structure and function of the RSV glycoproteins and their interactions with host cell factors that mediate entry. This period has coincided with renewed interest in developing effective interventions, including the isolation of potent monoclonal antibodies and small molecules and the design of novel vaccine candidates. In this Review, we summarize the recent findings that have begun to elucidate RSV entry mechanisms, describe progress on the development of new interventions and conclude with a perspective on gaps in our knowledge that require further investigation.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

Fig. 1: Respiratory syncytial virus virion.
Fig. 2: Attachment protein structure.
Fig. 3: Fusion protein structure.
Fig. 4: Attachment and fusion.
Fig. 5: Fusion protein binding sites for antibodies and small molecules.

References

  1. 1.

    Blount, R. E. Jr, Morris, J. A. & Savage, R. E. Recovery of cytopathogenic agent from chimpanzees with coryza. Proc. Soc. Exp. Biol. Med. 92, 544–549 (1956).

  2. 2.

    Chanock, R., Roizman, B. & Myers, R. Recovery from infants with respiratory illness of a virus related to chimpanzee coryza agent (CCA). I. Isolation, properties and characterization. Am. J. Hyg. 66, 281–290 (1957). This study reports the first isolation of RSV from infants.

  3. 3.

    Chanock, R. & Finberg, L. Recovery from infants with respiratory illness of a virus related to chimpanzee coryza agent (CCA). II. Epidemiologic aspects of infection in infants and young children. Am. J. Hyg. 66, 291–300 (1957).

  4. 4.

    Glezen, W. P., Taber, L. H., Frank, A. L. & Kasel, J. A. Risk of primary infection and reinfection with respiratory syncytial virus. Am. J. Dis. Child 140, 543–546 (1986).

  5. 5.

    Shi, T. et al. Global, regional, and national disease burden estimates of acute lower respiratory infections due to respiratory syncytial virus in young children in 2015: a systematic review and modelling study. Lancet 390, 946–958 (2017).

  6. 6.

    Falsey, A. R., Hennessey, P. A., Formica, M. A., Cox, C. & Walsh, E. E. Respiratory syncytial virus infection in elderly and high-risk adults. N. Engl. J. Med. 352, 1749–1759 (2005).

  7. 7.

    van den Hoogen, B. G. et al. A newly discovered human pneumovirus isolated from young children with respiratory tract disease. Nat. Med. 7, 719–724 (2001). This work describes the first isolation of human metapneumovirus from children.

  8. 8.

    Panda, S., Mohakud, N. K., Pena, L. & Kumar, S. Human metapneumovirus: review of an important respiratory pathogen. Int. J. Infect. Dis. 25, 45–52 (2014).

  9. 9.

    Hall, C. B. Respiratory syncytial virus: its transmission in the hospital environment. Yale J. Biol. Med. 55, 219–223 (1982).

  10. 10.

    Haas, L. E., Thijsen, S. F., van Elden, L. & Heemstra, K. A. Human metapneumovirus in adults. Viruses 5, 87–110 (2013).

  11. 11.

    Grayson, S. A., Griffiths, P. S., Perez, M. K. & Piedimonte, G. Detection of airborne respiratory syncytial virus in a pediatric acute care clinic. Pediatr. Pulmonol. 52, 684–688 (2017).

  12. 12.

    Collins, P. L. & Graham, B. S. Viral and host factors in human respiratory syncytial virus pathogenesis. J. Virol. 82, 2040–2055 (2008).

  13. 13.

    Peebles, R. S. Jr & Graham, B. S. Pathogenesis of respiratory syncytial virus infection in the murine model. Proc. Am. Thorac Soc. 2, 110–115 (2005).

  14. 14.

    Kim, H. W. et al. Respiratory syncytial virus disease in infants despite prior administration of antigenic inactivated vaccine. Am. J. Epidemiol. 89, 422–434 (1969).

  15. 15.

    Kapikian, A. Z., Mitchell, R. H., Chanock, R. M., Shvedoff, R. A. & Stewart, C. E. An epidemiologic study of altered clinical reactivity to respiratory syncytial (RS) virus infection in children previously vaccinated with an inactivated RS virus vaccine. Am. J. Epidemiol. 89, 405–421 (1969).

  16. 16.

    Fulginiti, V. A. et al. Respiratory virus immunization. I. A field trial of two inactivated respiratory virus vaccines; an aqueous trivalent parainfluenza virus vaccine and an alum-precipitated respiratory syncytial virus vaccine. Am. J. Epidemiol. 89, 435–448 (1969).

  17. 17.

    Chin, J., Magoffin, R. L., Shearer, L. A., Schieble, J. H. & Lennette, E. H. Field evaluation of a respiratory syncytial virus vaccine and a trivalent parainfluenza virus vaccine in a pediatric population. Am. J. Epidemiol. 89, 449–463 (1969).

  18. 18.

    Prince, G. A., Curtis, S. J., Yim, K. C. & Porter, D. D. Vaccine-enhanced respiratory syncytial virus disease in cotton rats following immunization with Lot 100 or a newly prepared reference vaccine. J. Gen. Virol. 82, 2881–2888 (2001).

  19. 19.

    Polack, F. P. et al. A role for immune complexes in enhanced respiratory syncytial virus disease. J. Exp. Med. 196, 859–865 (2002). This manuscript demonstrates that vaccine-enhanced disease is mediated by immune complexes.

  20. 20.

    Homaira, N., Rawlinson, W., Snelling, T. L. & Jaffe, A. Effectiveness of palivizumab in preventing RSV hospitalization in high risk children: a real-world perspective. Int. J. Pediatr. 2014, 571609 (2014).

  21. 21.

    Fearns, R. & Collins, P. L. Role of the M2-1 transcription antitermination protein of respiratory syncytial virus in sequential transcription. J. Virol. 73, 5852–5864 (1999).

  22. 22.

    Collins, P. L., Hill, M. G., Cristina, J. & Grosfeld, H. Transcription elongation factor of respiratory syncytial virus, a nonsegmented negative-strand RNA virus. Proc. Natl Acad. Sci. USA 93, 81–85 (1996).

  23. 23.

    Bermingham, A. & Collins, P. L. The M2-2 protein of human respiratory syncytial virus is a regulatory factor involved in the balance between RNA replication and transcription. Proc. Natl Acad. Sci. USA 96, 11259–11264 (1999).

  24. 24.

    Bitko, V. et al. Nonstructural proteins of respiratory syncytial virus suppress premature apoptosis by an NF-κB-dependent, interferon-independent mechanism and facilitate virus growth. J. Virol. 81, 1786–1795 (2007).

  25. 25.

    Spann, K. M., Tran, K. C. & Collins, P. L. Effects of nonstructural proteins NS1 and NS2 of human respiratory syncytial virus on interferon regulatory factor 3, NF-κB, and proinflammatory cytokines. J. Virol. 79, 5353–5362 (2005).

  26. 26.

    Gan, S. W. et al. The small hydrophobic protein of the human respiratory syncytial virus forms pentameric ion channels. J. Biol. Chem. 287, 24671–24689 (2012).

  27. 27.

    Fuentes, S., Tran, K. C., Luthra, P., Teng, M. N. & He, B. Function of the respiratory syncytial virus small hydrophobic protein. J. Virol. 81, 8361–8366 (2007).

  28. 28.

    Baviskar, P. S., Hotard, A. L., Moore, M. L. & Oomens, A. G. The respiratory syncytial virus fusion protein targets to the perimeter of inclusion bodies and facilitates filament formation by a cytoplasmic tail-dependent mechanism. J. Virol. 87, 10730–10741 (2013).

  29. 29.

    Shaikh, F. Y. et al. A critical phenylalanine residue in the respiratory syncytial virus fusion protein cytoplasmic tail mediates assembly of internal viral proteins into viral filaments and particles. mBio 3, e00270-11 (2012).

  30. 30.

    Oomens, A. G., Bevis, K. P. & Wertz, G. W. The cytoplasmic tail of the human respiratory syncytial virus F protein plays critical roles in cellular localization of the F protein and infectious progeny production. J. Virol. 80, 10465–10477 (2006).

  31. 31.

    Marty, A., Meanger, J., Mills, J., Shields, B. & Ghildyal, R. Association of matrix protein of respiratory syncytial virus with the host cell membrane of infected cells. Arch. Virol. 149, 199–210 (2004).

  32. 32.

    Kiss, G. et al. Structural analysis of respiratory syncytial virus reveals the position of M2-1 between the matrix protein and the ribonucleoprotein complex. J. Virol. 88, 7602–7617 (2014).

  33. 33.

    Tawar, R. G. et al. Crystal structure of a nucleocapsid-like nucleoprotein-RNA complex of respiratory syncytial virus. Science 326, 1279–1283 (2009).

  34. 34.

    Rixon, H. W. et al. The small hydrophobic (SH) protein accumulates within lipid-raft structures of the Golgi complex during respiratory syncytial virus infection. J. Gen. Virol. 85, 1153–1165 (2004).

  35. 35.

    Bukreyev, A., Whitehead, S. S., Murphy, B. R. & Collins, P. L. Recombinant respiratory syncytial virus from which the entire SH gene has been deleted grows efficiently in cell culture and exhibits site-specific attenuation in the respiratory tract of the mouse. J. Virol. 71, 8973–8982 (1997).

  36. 36.

    Levine, S., Klaiber-Franco, R. & Paradiso, P. R. Demonstration that glycoprotein G is the attachment protein of respiratory syncytial virus. J. Gen. Virol. 68, 2521–2524 (1987).

  37. 37.

    Hendricks, D. A., Baradaran, K., McIntosh, K. & Patterson, J. L. Appearance of a soluble form of the G protein of respiratory syncytial virus in fluids of infected cells. J. Gen. Virol. 68, 1705–1714 (1987).

  38. 38.

    Wertz, G. W. et al. Nucleotide sequence of the G protein gene of human respiratory syncytial virus reveals an unusual type of viral membrane protein. Proc. Natl Acad. Sci. USA 82, 4075–4079 (1985).

  39. 39.

    Collins, P. L. & Mottet, G. Oligomerization and post-translational processing of glycoprotein G of human respiratory syncytial virus: altered O-glycosylation in the presence of brefeldin A. J. Gen. Virol. 73, 849–863 (1992).

  40. 40.

    Satake, M., Coligan, J. E., Elango, N., Norrby, E. & Venkatesan, S. Respiratory syncytial virus envelope glycoprotein (G) has a novel structure. Nucleic Acids Res. 13, 7795–7812 (1985).

  41. 41.

    Garcia-Beato, R. et al. Host cell effect upon glycosylation and antigenicity of human respiratory syncytial virus G glycoprotein. Virology 221, 301–309 (1996).

  42. 42.

    Kwilas, S. et al. Respiratory syncytial virus grown in Vero cells contains a truncated attachment protein that alters its infectivity and dependence on glycosaminoglycans. J. Virol. 83, 10710–10718 (2009).

  43. 43.

    Roberts, S. R., Lichtenstein, D., Ball, L. A. & Wertz, G. W. The membrane-associated and secreted forms of the respiratory syncytial virus attachment glycoprotein G are synthesized from alternative initiation codons. J. Virol. 68, 4538–4546 (1994).

  44. 44.

    Hendricks, D. A., McIntosh, K. & Patterson, J. L. Further characterization of the soluble form of the G glycoprotein of respiratory syncytial virus. J. Virol. 62, 2228–2233 (1988).

  45. 45.

    Bukreyev, A., Yang, L. & Collins, P. L. The secreted G protein of human respiratory syncytial virus antagonizes antibody-mediated restriction of replication involving macrophages and complement. J. Virol. 86, 10880–10884 (2012).

  46. 46.

    Bukreyev, A. et al. The secreted form of respiratory syncytial virus G glycoprotein helps the virus evade antibody-mediated restriction of replication by acting as an antigen decoy and through effects on Fc receptor-bearing leukocytes. J. Virol. 82, 12191–12204 (2008).

  47. 47.

    Gorman, J. J., Ferguson, B. L., Speelman, D. & Mills, J. Determination of the disulfide bond arrangement of human respiratory syncytial virus attachment (G) protein by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Protein Sci. 6, 1308–1315 (1997).

  48. 48.

    Langedijk, J. P., Schaaper, W. M., Meloen, R. H. & van Oirschot, J. T. Proposed three-dimensional model for the attachment protein G of respiratory syncytial virus. J. Gen. Virol. 77, 1249–1257 (1996).

  49. 49.

    Doreleijers, J. F. et al. Solution structure of the immunodominant region of protein G of bovine respiratory syncytial virus. Biochemistry 35, 14684–14688 (1996). This study determines the first 3D structure of the RSV G cystine noose.

  50. 50.

    Sugawara, M. et al. Structure-antigenicity relationship studies of the central conserved region of human respiratory syncytial virus protein G. J. Pept. Res. 60, 271–282 (2002).

  51. 51.

    Langedijk, J. P., de Groot, B. L., Berendsen, H. J. & van Oirschot, J. T. Structural homology of the central conserved region of the attachment protein G of respiratory syncytial virus with the fourth subdomain of 55-kDa tumor necrosis factor receptor. Virology 243, 293–302 (1998).

  52. 52.

    Jones, H. G. et al. Structural basis for recognition of the central conserved region of RSV G by neutralizing human antibodies. PLOS Pathog. 14, e1006935 (2018).

  53. 53.

    Fedechkin, S. O., George, N. L., Wolff, J. T., Kauvar, L. M. & DuBois, R. M. Structures of respiratory syncytial virus G antigen bound to broadly neutralizing antibodies. Sci. Immunol. 3, eaar3534 (2018).

  54. 54.

    Pangesti, K. N. A., Abd El Ghany, M., Walsh, M. G., Kesson, A. M. & Hill-Cawthorne, G. A. Molecular epidemiology of respiratory syncytial virus. Rev. Med. Virol. 28, e1968 (2018).

  55. 55.

    Mufson, M. A., Orvell, C., Rafnar, B. & Norrby, E. Two distinct subtypes of human respiratory syncytial virus. J. Gen. Virol. 66, 2111–2124 (1985).

  56. 56.

    Anderson, L. J. et al. Antigenic characterization of respiratory syncytial virus strains with monoclonal antibodies. J. Infect. Dis. 151, 626–633 (1985).

  57. 57.

    Hall, C. B. et al. Occurrence of groups A and B of respiratory syncytial virus over 15 years: associated epidemiologic and clinical characteristics in hospitalized and ambulatory children. J. Infect. Dis. 162, 1283–1290 (1990).

  58. 58.

    Trento, A. et al. Major changes in the G protein of human respiratory syncytial virus isolates introduced by a duplication of 60 nucleotides. J. Gen. Virol. 84, 3115–3120 (2003).

  59. 59.

    Trento, A. et al. Natural history of human respiratory syncytial virus inferred from phylogenetic analysis of the attachment (G) glycoprotein with a 60-nucleotide duplication. J. Virol. 80, 975–984 (2006).

  60. 60.

    Trento, A. et al. Ten years of global evolution of the human respiratory syncytial virus BA genotype with a 60-nucleotide duplication in the G protein gene. J. Virol. 84, 7500–7512 (2010).

  61. 61.

    Eshaghi, A. et al. Genetic variability of human respiratory syncytial virus A strains circulating in Ontario: a novel genotype with a 72 nucleotide G gene duplication. PLOS ONE 7, e32807 (2012).

  62. 62.

    Duvvuri, V. R. et al. Genetic diversity and evolutionary insights of respiratory syncytial virus A ON1 genotype: global and local transmission dynamics. Sci. Rep. 5, 14268 (2015).

  63. 63.

    Hirano, E. et al. Molecular evolution of human respiratory syncytial virus attachment glycoprotein (G) gene of new genotype ON1 and ancestor NA1. Infect. Genet. Evol. 28, 183–191 (2014).

  64. 64.

    Hotard, A. L., Laikhter, E., Brooks, K., Hartert, T. V. & Moore, M. L. Functional analysis of the 60-nucleotide duplication in the respiratory syncytial virus buenos aires strain attachment glycoprotein. J. Virol. 89, 8258–8266 (2015).

  65. 65.

    Leyrat, C., Paesen, G. C., Charleston, J., Renner, M. & Grimes, J. M. Structural insights into the human metapneumovirus glycoprotein ectodomain. J. Virol. 88, 11611–11616 (2014).

  66. 66.

    Yin, H. S., Wen, X., Paterson, R. G., Lamb, R. A. & Jardetzky, T. S. Structure of the parainfluenza virus 5 F protein in its metastable, prefusion conformation. Nature 439, 38–44 (2006).

  67. 67.

    Yin, H. S., Paterson, R. G., Wen, X., Lamb, R. A. & Jardetzky, T. S. Structure of the uncleaved ectodomain of the paramyxovirus (hPIV3) fusion protein. Proc. Natl Acad. Sci. USA 102, 9288–9293 (2005).

  68. 68.

    Collins, P. L., Huang, Y. T. & Wertz, G. W. Nucleotide sequence of the gene encoding the fusion (F) glycoprotein of human respiratory syncytial virus. Proc. Natl Acad. Sci. USA 81, 7683–7687 (1984).

  69. 69.

    Zimmer, G., Budz, L. & Herrler, G. Proteolytic activation of respiratory syncytial virus fusion protein. Cleavage at two furin consensus sequences. J. Biol. Chem. 276, 31642–31650 (2001).

  70. 70.

    Gonzalez-Reyes, L. et al. Cleavage of the human respiratory syncytial virus fusion protein at two distinct sites is required for activation of membrane fusion. Proc. Natl Acad. Sci. USA 98, 9859–9864 (2001).

  71. 71.

    Bolt, G., Pedersen, L. O. & Birkeslund, H. H. Cleavage of the respiratory syncytial virus fusion protein is required for its surface expression: role of furin. Virus Res. 68, 25–33 (2000).

  72. 72.

    Collins, P. L. & Mottet, G. Post-translational processing and oligomerization of the fusion glycoprotein of human respiratory syncytial virus. J. Gen. Virol. 72, 3095–3101 (1991).

  73. 73.

    Day, N. D. et al. Contribution of cysteine residues in the extracellular domain of the F protein of human respiratory syncytial virus to its function. Virol. J. 3, 34 (2006).

  74. 74.

    Gilman, M. S. et al. Characterization of a prefusion-specific antibody that recognizes a quaternary, cleavage-dependent epitope on the RSV fusion glycoprotein. PLOS Pathog. 11, e1005035 (2015).

  75. 75.

    Krarup, A. et al. A highly stable prefusion RSV F vaccine derived from structural analysis of the fusion mechanism. Nat. Commun. 6, 8143 (2015).

  76. 76.

    McLellan, J. S. et al. Structure of RSV fusion glycoprotein trimer bound to a prefusion-specific neutralizing antibody. Science 340, 1113–1117 (2013). This work provides the first 3D structure of the prefusion conformation of RSV F and defines a major antigenic site recognized by prefusion-specific antibodies.

  77. 77.

    Liljeroos, L., Krzyzaniak, M. A., Helenius, A. & Butcher, S. J. Architecture of respiratory syncytial virus revealed by electron cryotomography. Proc. Natl Acad. Sci. USA 110, 11133–11138 (2013). This manuscript reveals the organization and morphology of RSV virions by cryo-electron tomography.

  78. 78.

    Killikelly, A. M., Kanekiyo, M. & Graham, B. S. Pre-fusion F is absent on the surface of formalin-inactivated respiratory syncytial virus. Sci. Rep. 6, 34108 (2016).

  79. 79.

    Kim, Y. H. et al. Capture and imaging of a prehairpin fusion intermediate of the paramyxovirus PIV5. Proc. Natl Acad. Sci. USA 108, 20992–20997 (2011).

  80. 80.

    Zhao, X., Singh, M., Malashkevich, V. N. & Kim, P. S. Structural characterization of the human respiratory syncytial virus fusion protein core. Proc. Natl Acad. Sci. USA 97, 14172–14177 (2000).

  81. 81.

    McLellan, J. S., Yang, Y., Graham, B. S. & Kwong, P. D. Structure of respiratory syncytial virus fusion glycoprotein in the postfusion conformation reveals preservation of neutralizing epitopes. J. Virol. 85, 7788–7796 (2011).

  82. 82.

    Swanson, K. A. et al. Structural basis for immunization with postfusion respiratory syncytial virus fusion F glycoprotein (RSV F) to elicit high neutralizing antibody titers. Proc. Natl Acad. Sci. USA 108, 9619–9624 (2011).

  83. 83.

    Johnson, J. E., Gonzales, R. A., Olson, S. J., Wright, P. F. & Graham, B. S. The histopathology of fatal untreated human respiratory syncytial virus infection. Mod. Pathol. 20, 108–119 (2007).

  84. 84.

    Xu, L. et al. A fatal case associated with respiratory syncytial virus infection in a young child. BMC Infect. Dis. 18, 217 (2018).

  85. 85.

    Pitkaranta, A., Virolainen, A., Jero, J., Arruda, E. & Hayden, F. G. Detection of rhinovirus, respiratory syncytial virus, and coronavirus infections in acute otitis media by reverse transcriptase polymerase chain reaction. Pediatrics 102, 291–295 (1998).

  86. 86.

    Rohwedder, A. et al. Detection of respiratory syncytial virus RNA in blood of neonates by polymerase chain reaction. J. Med. Virol. 54, 320–327 (1998).

  87. 87.

    Escribano-Romero, E., Rawling, J., Garcia-Barreno, B. & Melero, J. A. The soluble form of human respiratory syncytial virus attachment protein differs from the membrane-bound form in its oligomeric state but is still capable of binding to cell surface proteoglycans. J. Virol. 78, 3524–3532 (2004).

  88. 88.

    Krusat, T. & Streckert, H. J. Heparin-dependent attachment of respiratory syncytial virus (RSV) to host cells. Arch. Virol. 142, 1247–1254 (1997).

  89. 89.

    Feldman, S. A., Hendry, R. M. & Beeler, J. A. Identification of a linear heparin binding domain for human respiratory syncytial virus attachment glycoprotein G. J. Virol. 73, 6610–6617 (1999).

  90. 90.

    Hallak, L. K., Spillmann, D., Collins, P. L. & Peeples, M. E. Glycosaminoglycan sulfation requirements for respiratory syncytial virus infection. J. Virol. 74, 10508–10513 (2000).

  91. 91.

    Martinez, I. & Melero, J. A. Binding of human respiratory syncytial virus to cells: implication of sulfated cell surface proteoglycans. J. Gen. Virol. 81, 2715–2722 (2000).

  92. 92.

    Hallak, L. K., Collins, P. L., Knudson, W. & Peeples, M. E. Iduronic acid-containing glycosaminoglycans on target cells are required for efficient respiratory syncytial virus infection. Virology 271, 264–275 (2000).

  93. 93.

    Chirkova, T. et al. CX3CR1 is an important surface molecule for respiratory syncytial virus infection in human airway epithelial cells. J. Gen. Virol. 96, 2543–2556 (2015).

  94. 94.

    Zhang, L. et al. Infection of ciliated cells by human parainfluenza virus type 3 in an in vitro model of human airway epithelium. J. Virol. 79, 1113–1124 (2005).

  95. 95.

    Zhang, L., Peeples, M. E., Boucher, R. C., Collins, P. L. & Pickles, R. J. Respiratory syncytial virus infection of human airway epithelial cells is polarized, specific to ciliated cells, and without obvious cytopathology. J. Virol. 76, 5654–5666 (2002).

  96. 96.

    Johnson, S. M. et al. Respiratory syncytial virus uses CX3CR1 as a receptor on primary human airway epithelial cultures. PLOS Pathog. 11, e1005318 (2015).

  97. 97.

    Tripp, R. A. et al. CX3C chemokine mimicry by respiratory syncytial virus G glycoprotein. Nat. Immunol. 2, 732–738 (2001). This study identifies the CX 3 C motif in RSV G and demonstrates that CX 3 CR1 facilitates RSV entry.

  98. 98.

    Bazan, J. F. et al. A new class of membrane-bound chemokine with a CX3C motif. Nature 385, 640–644 (1997).

  99. 99.

    Jeong, K. I. et al. CX3CR1 is expressed in differentiated human ciliated airway cells and co-localizes with respiratory syncytial virus on cilia in a G protein-dependent manner. PLOS ONE 10, e0130517 (2015).

  100. 100.

    Karron, R. A. et al. Respiratory syncytial virus (RSV) SH and G proteins are not essential for viral replication in vitro: clinical evaluation and molecular characterization of a cold-passaged, attenuated RSV subgroup B mutant. Proc. Natl Acad. Sci. USA 94, 13961–13966 (1997). This work demonstrates that infectious RSV requires only the F protein on its surface.

  101. 101.

    Techaarpornkul, S., Barretto, N. & Peeples, M. E. Functional analysis of recombinant respiratory syncytial virus deletion mutants lacking the small hydrophobic and/or attachment glycoprotein gene. J. Virol. 75, 6825–6834 (2001).

  102. 102.

    Feldman, S. A., Audet, S. & Beeler, J. A. The fusion glycoprotein of human respiratory syncytial virus facilitates virus attachment and infectivity via an interaction with cellular heparan sulfate. J. Virol. 74, 6442–6447 (2000).

  103. 103.

    Behera, A. K. et al. Blocking intercellular adhesion molecule-1 on human epithelial cells decreases respiratory syncytial virus infection. Biochem. Biophys. Res. Commun. 280, 188–195 (2001).

  104. 104.

    Currier, M. G. et al. EGFR interacts with the fusion protein of respiratory syncytial virus strain 2–20 and mediates infection and mucin expression. PLOS Pathog. 12, e1005622 (2016).

  105. 105.

    Tayyari, F. et al. Identification of nucleolin as a cellular receptor for human respiratory syncytial virus. Nat. Med. 17, 1132–1135 (2011). This manuscript identifies nucleolin as a host cell factor that interacts with the F protein and facilitates RSV entry.

  106. 106.

    Bose, S., Basu, M. & Banerjee, A. K. Role of nucleolin in human parainfluenza virus type 3 infection of human lung epithelial cells. J. Virol. 78, 8146–8158 (2004).

  107. 107.

    Su, P. Y. et al. Cell surface nucleolin facilitates enterovirus 71 binding and infection. J. Virol. 89, 4527–4538 (2015).

  108. 108.

    Xiao, X., Feng, Y., Zhu, Z. & Dimitrov, D. S. Identification of a putative Crimean-Congo hemorrhagic fever virus entry factor. Biochem. Biophys. Res. Commun. 411, 253–258 (2011).

  109. 109.

    Qiu, J. & Brown, K. E. A. 110-kDa nuclear shuttle protein, nucleolin, specifically binds to adeno-associated virus type 2 (AAV-2) capsid. Virology 257, 373–382 (1999).

  110. 110.

    Callebaut, C. et al. Identification of V3 loop-binding proteins as potential receptors implicated in the binding of HIV particles to CD4+ cells. J. Biol. Chem. 273, 21988–21997 (1998).

  111. 111.

    Srinivasakumar, N., Ogra, P. L. & Flanagan, T. D. Characteristics of fusion of respiratory syncytial virus with HEp-2 cells as measured by R18 fluorescence dequenching assay. J. Virol. 65, 4063–4069 (1991).

  112. 112.

    Kahn, J. S., Schnell, M. J., Buonocore, L. & Rose, J. K. Recombinant vesicular stomatitis virus expressing respiratory syncytial virus (RSV) glycoproteins: RSV fusion protein can mediate infection and cell fusion. Virology 254, 81–91 (1999).

  113. 113.

    White, J. M. & Whittaker, G. R. Fusion of enveloped viruses in endosomes. Traffic 17, 593–614 (2016).

  114. 114.

    San-Juan-Vergara, H. et al. Cholesterol-rich microdomains as docking platforms for respiratory syncytial virus in normal human bronchial epithelial cells. J. Virol. 86, 1832–1843 (2012).

  115. 115.

    Krzyzaniak, M. A., Zumstein, M. T., Gerez, J. A., Picotti, P. & Helenius, A. Host cell entry of respiratory syncytial virus involves macropinocytosis followed by proteolytic activation of the F protein. PLOS Pathog. 9, e1003309 (2013).

  116. 116.

    Schlender, J., Zimmer, G., Herrler, G. & Conzelmann, K. K. Respiratory syncytial virus (RSV) fusion protein subunit F2, not attachment protein G, determines the specificity of RSV infection. J. Virol. 77, 4609–4616 (2003).

  117. 117.

    Yuan, P. et al. Structural studies of the parainfluenza virus 5 hemagglutinin-neuraminidase tetramer in complex with its receptor, sialyllactose. Structure 13, 803–815 (2005).

  118. 118.

    Crennell, S., Takimoto, T., Portner, A. & Taylor, G. Crystal structure of the multifunctional paramyxovirus hemagglutinin-neuraminidase. Nat. Struct. Biol. 7, 1068–1074 (2000).

  119. 119.

    Bose, S., Jardetzky, T. S. & Lamb, R. A. Timing is everything: fine-tuned molecular machines orchestrate paramyxovirus entry. Virology 479–480, 518–531 (2015).

  120. 120.

    Yunus, A. S. et al. Elevated temperature triggers human respiratory syncytial virus F protein six-helix bundle formation. Virology 396, 226–237 (2010).

  121. 121.

    Fearns, R. & Deval, J. New antiviral approaches for respiratory syncytial virus and other mononegaviruses: inhibiting the RNA polymerase. Antiviral Res. 134, 63–76 (2016).

  122. 122.

    Falsey, A. R. & Walsh, E. E. Relationship of serum antibody to risk of respiratory syncytial virus infection in elderly adults. J. Infect. Dis. 177, 463–466 (1998).

  123. 123.

    Hall, C. B., Walsh, E. E., Long, C. E. & Schnabel, K. C. Immunity to and frequency of reinfection with respiratory syncytial virus. J. Infect. Dis. 163, 693–698 (1991).

  124. 124.

    American Academy of Pediatrics. Respiratory syncytial virus immune globulin intravenous: indications for use. Committee on Infectious Diseases, Committee on Fetus and Newborn. Pediatrics 99, 645–650 (1997).

  125. 125.

    The IMpact-RSV Study Group. Palivizumab, a humanized respiratory syncytial virus monoclonal antibody, reduces hospitalization from respiratory syncytial virus infection in high-risk infants. Pediatrics 102, 531–537 (1998).

  126. 126.

    Beeler, J. A. & Coelingh, K. V. Neutralization epitopes of the F-glycoprotein of respiratory syncytial virus - effect of mutation upon fusion function. J. Virol. 63, 2941–2950 (1989).

  127. 127.

    Kwakkenbos, M. J. et al. Generation of stable monoclonal antibody-producing B cell receptor-positive human memory B cells by genetic programming. Nat. Med. 16, 123–128 (2010). This study reports the isolation and characterization of the first prefusion F-specific monoclonal antibodies, although their specificity was not known at the time.

  128. 128.

    Goodwin, E. et al. Infants infected with respiratory syncytial virus generate potent neutralizing antibodies that lack somatic hypermutation. Immunity 48, 339–349 (2018).

  129. 129.

    Gilman, M. S. et al. Rapid profiling of RSV antibody repertoires from the memory B cells of naturally infected adult donors. Sci. Immunol. 1 (2016).

  130. 130.

    Collarini, E. J. et al. Potent high-affinity antibodies for treatment and prophylaxis of respiratory syncytial virus derived from B cells of infected patients. J. Immunol. 183, 6338–6345 (2009).

  131. 131.

    Mousa, J. J., Kose, N., Matta, P., Gilchuk, P. & Crowe, J. E. Jr. A novel pre-fusion conformation-specific neutralizing epitope on the respiratory syncytial virus fusion protein. Nat. Microbiol. 2, 16271 (2017).

  132. 132.

    Corti, D. et al. Cross-neutralization of four paramyxoviruses by a human monoclonal antibody. Nature 501, 439–443 (2013).

  133. 133.

    Zhu, Q. et al. A highly potent extended half-life antibody as a potential RSV vaccine surrogate for all infants. Sci. Transl Med. 9, eaaj1928 (2017).

  134. 134.

    US National Library of Medicine. ClinicalTrials.gov https://clinicaltrials.gov/ct2/show/NCT02878330 (2018).

  135. 135.

    US National Library of Medicine. ClinicalTrials.gov https://clinicaltrials.gov/ct2/show/NCT02325791 (2018).

  136. 136.

    Costello, H. M., Ray, W. C., Chaiwatpongsakorn, S. & Peeples, M. E. Targeting RSV with vaccines and small molecule drugs. Infect. Disord. Drug Targets 12, 110–128 (2012).

  137. 137.

    Heylen, E., Neyts, J. & Jochmans, D. Drug candidates and model systems in respiratory syncytial virus antiviral drug discovery. Biochem. Pharmacol. 127, 1–12 (2017).

  138. 138.

    Cianci, C. et al. Targeting a binding pocket within the trimer-of-hairpins: small-molecule inhibition of viral fusion. Proc. Natl Acad. Sci. USA 101, 15046–15051 (2004).

  139. 139.

    Roymans, D. et al. Binding of a potent small-molecule inhibitor of six-helix bundle formation requires interactions with both heptad-repeats of the RSV fusion protein. Proc. Natl Acad. Sci. USA 107, 308–313 (2010).

  140. 140.

    Yan, D. et al. Cross-resistance mechanism of respiratory syncytial virus against structurally diverse entry inhibitors. Proc. Natl Acad. Sci. USA 111, E3441–E3449 (2014).

  141. 141.

    Battles, M. B. et al. Molecular mechanism of respiratory syncytial virus fusion inhibitors. Nat. Chem. Biol. 12, 87–93 (2016). This work describes the binding site and mechanism of action for small-molecule fusion inhibitors.

  142. 142.

    Samuel, D. et al. GS-5806 inhibits pre- to postfusion conformational changes of the respiratory syncytial virus fusion protein. Antimicrob. Agents Chemother. 59, 7109–7112 (2015).

  143. 143.

    DeVincenzo, J. P. et al. Oral GS-5806 activity in a respiratory syncytial virus challenge study. N. Engl. J. Med. 371, 711–722 (2014).

  144. 144.

    Stevens, M. et al. Antiviral activity of oral JNJ-53718678 in healthy adult volunteers challenged with respiratory syncytial virus: a placebo-controlled study. J. Infect. Dis. 218, 748–756 (2018).

  145. 145.

    Mazur, N. I. et al. The respiratory syncytial virus vaccine landscape: lessons from the graveyard and promising candidates. Lancet Infect. Dis. 18, e295–e311 (2018).

  146. 146.

    Graham, B. S. Vaccine development for respiratory syncytial virus. Curr. Opin. Virol. 23, 107–112 (2017).

  147. 147.

    Magro, M. et al. Neutralizing antibodies against the preactive form of respiratory syncytial virus fusion protein offer unique possibilities for clinical intervention. Proc. Natl Acad. Sci. USA 109, 3089–3094 (2012). This study provides the first evidence for the existence of prefusion F-specific antibodies and their dominant contribution to the RSV-neutralizing activity of human sera.

  148. 148.

    Ngwuta, J. O. et al. Prefusion F-specific antibodies determine the magnitude of RSV neutralizing activity in human sera. Sci. Transl Med. 7, 309ra162 (2015).

  149. 149.

    McLellan, J. S. et al. Structure-based design of a fusion glycoprotein vaccine for respiratory syncytial virus. Science 342, 592–598 (2013). This manuscript reports the first structure-based design of a prefusion F vaccine antigen and demonstrates its superior immunogenicity to postfusion F antigens.

  150. 150.

    US National Library of Medicine. ClinicalTrials.gov https://clinicaltrials.gov/ct2/show/NCT03049488 (2018).

  151. 151.

    US National Library of Medicine. ClinicalTrials.gov https://clinicaltrials.gov/ct2/show/NCT03529773 (2018).

  152. 152.

    Falloon, J. et al. An adjuvanted, postfusion F protein-based vaccine did not prevent respiratory syncytial virus illness in older adults. J. Infect. Dis. 216, 1362–1370 (2017).

  153. 153.

    US National Library of Medicine. ClinicalTrials.gov https://clinicaltrials.gov/ct2/show/NCT02608502 (2017).

  154. 154.

    US National Library of Medicine. ClinicalTrials.gov https://clinicaltrials.gov/ct2/show/NCT02508194 (2017).

  155. 155.

    US National Library of Medicine. ClinicalTrials.gov https://clinicaltrials.gov/ct2/show/NCT02624947 (2018).

  156. 156.

    Karron, R. A., Buchholz, U. J. & Collins, P. L. Live-attenuated respiratory syncytial virus vaccines. Curr. Top. Microbiol. Immunol. 372, 259–284 (2013).

  157. 157.

    Karron, R. A. et al. A gene deletion that up-regulates viral gene expression yields an attenuated RSV vaccine with improved antibody responses in children. Sci. Transl Med. 7, 312ra175 (2015).

  158. 158.

    Liang, B. et al. Improved prefusion stability, optimized codon usage, and augmented virion packaging enhance the immunogenicity of respiratory syncytial virus fusion protein in a vectored-vaccine candidate. J. Virol. 91, e00189-17 (2017).

  159. 159.

    Stobart, C. C. et al. A live RSV vaccine with engineered thermostability is immunogenic in cotton rats despite high attenuation. Nat. Commun. 7, 13916 (2016).

  160. 160.

    US National Library of Medicine. ClinicalTrials.gov https://clinicaltrials.gov/ct2/show/NCT03303625 (2018).

  161. 161.

    US National Library of Medicine. ClinicalTrials.gov https://clinicaltrials.gov/ct2/show/NCT02927873 (2018).

  162. 162.

    US National Library of Medicine. ClinicalTrials.gov https://clinicaltrials.gov/ct2/show/NCT02873286 (2018).

  163. 163.

    Levine, S. Polypeptides of respiratory syncytial virus. J. Virol. 21, 427–431 (1977).

  164. 164.

    Walsh, E. E. & Hruska, J. Monoclonal antibodies to respiratory syncytial virus proteins: identification of the fusion protein. J. Virol. 47, 171–177 (1983).

  165. 165.

    Garcia, J., Garcia-Barreno, B., Vivo, A. & Melero, J. A. Cytoplasmic inclusions of respiratory syncytial virus-infected cells: formation of inclusion bodies in transfected cells that coexpress the nucleoprotein, the phosphoprotein, and the 22K protein. Virology 195, 243–247 (1993).

  166. 166.

    Garcia-Barreno, B., Delgado, T. & Melero, J. A. Identification of protein regions involved in the interaction of human respiratory syncytial virus phosphoprotein and nucleoprotein: significance for nucleocapsid assembly and formation of cytoplasmic inclusions. J. Virol. 70, 801–808 (1996).

  167. 167.

    Rincheval, V. et al. Functional organization of cytoplasmic inclusion bodies in cells infected by respiratory syncytial virus. Nat. Commun. 8, 563 (2017).

  168. 168.

    Noton, S. L. & Fearns, R. Initiation and regulation of paramyxovirus transcription and replication. Virology 479–480, 545–554 (2015).

  169. 169.

    Gower, T. L. et al. RhoA signaling is required for respiratory syncytial virus-induced syncytium formation and filamentous virion morphology. J. Virol. 79, 5326–5336 (2005).

  170. 170.

    Ke, Z. et al. The morphology and assembly of respiratory syncytial virus revealed by cryo-electron tomography. Viruses 10, E446 (2018). This work conclusively demonstrates that RSV is a filamentous virus upon budding from infected cells.

  171. 171.

    Mehedi, M. et al. Actin-related protein 2 (ARP2) and virus-induced filopodia facilitate human respiratory syncytial virus spread. PLOS Pathog. 12, e1006062 (2016).

  172. 172.

    Vanover, D. et al. RSV glycoprotein and genomic RNA dynamics reveal filament assembly prior to the plasma membrane. Nat. Commun. 8, 667 (2017).

  173. 173.

    Forster, A., Maertens, G. N., Farrell, P. J. & Bajorek, M. Dimerization of matrix protein is required for budding of respiratory syncytial virus. J. Virol. 89, 4624–4635 (2015).

  174. 174.

    Roberts, S. R., Compans, R. W. & Wertz, G. W. Respiratory syncytial virus matures at the apical surfaces of polarized epithelial cells. J. Virol. 69, 2667–2673 (1995).

  175. 175.

    Jardetzky, T. S. & Lamb, R. A. Activation of paramyxovirus membrane fusion and virus entry. Curr. Opin. Virol. 5, 24–33 (2014).

  176. 176.

    Yuan, P. et al. Structure of the Newcastle disease virus hemagglutinin-neuraminidase (HN) ectodomain reveals a four-helix bundle stalk. Proc. Natl Acad. Sci. USA 108, 14920–14925 (2011).

  177. 177.

    Welch, B. D. et al. Structure of the parainfluenza virus 5 (PIV5) hemagglutinin-neuraminidase (HN) ectodomain. PLOS Pathog. 9, e1003534 (2013).

  178. 178.

    Bose, S. et al. Fusion activation by a headless parainfluenza virus 5 hemagglutinin-neuraminidase stalk suggests a modular mechanism for triggering. Proc. Natl Acad. Sci. USA 109, E2625–E2634 (2012).

  179. 179.

    Brindley, M. A. et al. A stabilized headless measles virus attachment protein stalk efficiently triggers membrane fusion. J. Virol. 87, 11693–11703 (2013).

  180. 180.

    Liu, Q. et al. Unraveling a three-step spatiotemporal mechanism of triggering of receptor-induced Nipah virus fusion and cell entry. PLOS Pathog. 9, e1003770 (2013).

  181. 181.

    Iorio, R. M., Melanson, V. R. & Mahon, P. J. Glycoprotein interactions in paramyxovirus fusion. Future Virol. 4, 335–351 (2009).

  182. 182.

    McLellan, J. S. Neutralizing epitopes on the respiratory syncytial virus fusion glycoprotein. Curr. Opin. Virol. 11, 70–75 (2015).

Download references

Acknowledgements

The authors dedicate this Review to the memory of José A. Melero, a wonderful colleague and scientist who contributed much to the study of the RSV F and G proteins. The authors thank B. Graham, J. Langedijk and members of the McLellan laboratory for helpful comments on the manuscript, and M. Gilman for assistance with the figures.

Reviewer information

Nature Reviews Microbiology thanks L. Bont and other anonymous reviewer(s) for their contribution to the peer review of this work.

Author information

M.B.B. researched data for the article. M.B.B. and J.S.M. made substantial contributions to discussions of the content. M.B.B. and J.S.M. wrote the article. J.S.M. reviewed and edited the manuscript before submission.

Correspondence to Jason S. McLellan.

Ethics declarations

Competing interests

J.S.M. is a named inventor on patents for vaccines and/or monoclonal antibodies for RSV and coronaviruses, has received research funding from MedImmune and Janssen Pharmaceuticals, has been a paid consultant for MedImmune and is on the scientific advisory board for Calder Biosciences. M.B.B. is currently employed by Adimab.

Additional information

Publisher’s note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Glossary

Nasopharynx

The upper part of the pharynx that connects with the nasal cavity.

Bronchioles

Small tubes in the lung through which air is delivered to the alveoli.

Alveoli

Small air sacs in the lung that provide rapid gas exchange with blood.

Bronchiolitis

Inflammation of the bronchioles that reduces air passage.

Formalin

An aqueous solution of formaldehyde.

Neutrophil

Most abundant type of white blood cell.

Immune complex

An antibody bound to its antigen.

Passive immunoprophylaxis

The administration of an exogenously produced antibody given before infection occurs.

Apoptosis

Programmed cell death.

Glycoproteins

Proteins to which carbohydrates are covalently attached.

Ectodomain

The portion of a membrane protein that resides outside the cell or virion.

Cystine noose

A surface-accessible loop structure containing one or more disulfide bonds.

Serotype

A serologically distinguishable strain of a microorganism.

Protomer

A structural unit of an oligomeric protein.

Heptad repeat

A seven-amino-acid motif ‘abcdefg’ where a and d are hydrophobic.

Antigenic drift

The accumulation of amino acid substitutions that reduce antibody binding.

Apical surface

The surface of a polarized cell that faces the lumen or external environment.

Type 1 alveolar pneumocytes

Surface epithelial cells of alveoli involved in gas exchange.

Chemokine

A small secreted protein that stimulates recruitment of white blood cells.

Macropinocytosis

The nonselective uptake of extracellular molecules into endocytic vesicles.

Bronchopulmonary dysplasia

A chronic lung disease caused by mechanical ventilation and long-term oxygen use that results in damage to alveoli.

Antigenic site

A group of spatially related antibody epitopes.

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Further reading