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Abstract

Replication models have been developed to describe the replication dynamics of a variety of viruses to better understand the kinetics and key contributing factors affecting infectivity and spread. However, accurate representations of the dynamics of virus replication observed and are often limited due to the failure of these models to account for both environmental influences, such as temperature, and the variety of possible mechanisms employed by viruses to spread. Several major families of viruses including paramyxoviruses, pneumoviruses and coronaviruses, induce and use the formation of syncytia, large multinucleated cell masses formed through fusion of cells, to aid in spread to neighbouring susceptible cells. In this study, we evaluate and compare both the dynamics and roles of temperature and syncytia formation on the replication of two different fusogenic viruses : human respiratory syncytial virus (hRSV) and a murine coronavirus, mouse hepatitis virus (MHV). Thermal stability, replication kinetics and both the rates and dynamics of syncytia formation were evaluated for hRSV and MHV. These data were then incorporated into a novel and improved replication model for each of the two viruses, which provides new insights into the contributions of both temperature and syncytia formation in the replication of fusogenic viruses.

Funding
This study was supported by the:
  • Holcomb Awards Committee - Butler University (Award 028657)
    • Principal Award Recipient: ChristopherC. Stobart
  • National Science Foundation (Award 2116348)
    • Principal Award Recipient: LindsayLewellyn
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/content/journal/jgv/10.1099/jgv.0.002078
2025-02-13
2026-03-12

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References

  1. Cifuentes-Munoz N, El Najjar F, Dutch RE. Viral cell-to-cell spread: conventional and non-conventional ways. Adv Virus Res 2020; 108:85–125 [View Article] [PubMed]
    [Google Scholar]
  2. Leroy H, Han M, Woottum M, Bracq L, Bouchet J et al. Virus-mediated cell-cell fusion. Int J Mol Sci 2020; 21:9644 [View Article] [PubMed]
    [Google Scholar]
  3. Zhong P, Agosto LM, Munro JB, Mothes W. Cell-to-cell transmission of viruses. Curr Opin Virol 2013; 3:44–50 [View Article] [PubMed]
    [Google Scholar]
  4. Sattentau Q. Avoiding the void: cell-to-cell spread of human viruses. Nat Rev Microbiol 2008; 6:815–826 [View Article] [PubMed]
    [Google Scholar]
  5. Gamble A, Yeo YY, Butler AA, Tang H, Snedden CE et al. Drivers and distribution of henipavirus-induced syncytia: what do we know?. Viruses 2021; 13:1755 [View Article] [PubMed]
    [Google Scholar]
  6. Symeonides M, Murooka TT, Bellfy LN, Roy NH, Mempel TR et al. HIV-1-induced small T cell syncytia can transfer virus particles to target cells through transient contacts. Viruses 2015; 7:6590–6603 [View Article] [PubMed]
    [Google Scholar]
  7. Oliver SL, Zhou M, Arvin AM. Varicella-zoster virus: molecular controls of cell fusion-dependent pathogenesis. Biochem Soc Trans 2020; 48:2415–2435 [View Article] [PubMed]
    [Google Scholar]
  8. Connolly SA, Jardetzky TS, Longnecker R. The structural basis of herpesvirus entry. Nat Rev Microbiol 2021; 19:110–121 [View Article] [PubMed]
    [Google Scholar]
  9. Bracq L, Xie M, Benichou S, Bouchet J. Mechanisms for cell-to-cell transmission of HIV-1. Front Immunol 2018; 9:260 [View Article] [PubMed]
    [Google Scholar]
  10. Lavi E, Wang Q, Weiss SR, Gonatas NK. Syncytia formation induced by coronavirus infection is associated with fragmentation and rearrangement of the Golgi apparatus. Virology 1996; 221:325–334 [View Article] [PubMed]
    [Google Scholar]
  11. Fenrich M, Mrdenovic S, Balog M, Tomic S, Zjalic M et al. SARS-CoV-2 dissemination through peripheral nerves explains multiple organ injury. Front Cell Neurosci 2020; 14:229 [View Article] [PubMed]
    [Google Scholar]
  12. Belouzard S, Millet JK, Licitra BN, Whittaker GR. Mechanisms of coronavirus cell entry mediated by the viral spike protein. Viruses 2012; 4:1011–1033 [View Article] [PubMed]
    [Google Scholar]
  13. Chang A, Dutch RE. Paramyxovirus fusion and entry: multiple paths to a common end. Viruses 2012; 4:613–636 [View Article] [PubMed]
    [Google Scholar]
  14. Freeman MC, Graham RL, Lu X, Peek CT, Denison MR. Coronavirus replicase-reporter fusions provide quantitative analysis of replication and replication complex formation. J Virol 2014; 88:5319–5327 [View Article] [PubMed]
    [Google Scholar]
  15. Wang Y, Grunewald M, Perlman S. Coronaviruses: an updated overview of their replication and pathogenesis. Methods Mol Biol Clifton NJ 2020; 2203:1–29
    [Google Scholar]
  16. Harrison CM, Doster JM, Landwehr EH, Kumar NP, White EJ et al. Evaluating the virology and evolution of seasonal human coronaviruses associated with the common cold in the COVID-19 era. Microorganisms 2023; 11:445 [View Article] [PubMed]
    [Google Scholar]
  17. Lee N, Hui D, Wu A, Chan P, Cameron P et al. A major outbreak of severe acute respiratory syndrome in Hong Kong. N Engl J Med 2003; 348:1986–1994 [View Article] [PubMed]
    [Google Scholar]
  18. Peiris JSM, Yuen KY, Osterhaus ADME, Stöhr K. The severe acute respiratory syndrome. N Engl J Med 2003; 349:2431–2441 [View Article]
    [Google Scholar]
  19. Zaki AM, van Boheemen S, Bestebroer TM, Osterhaus ADME, Fouchier RAM. Isolation of a novel coronavirus from a man with pneumonia in Saudi Arabia. N Engl J Med 2012; 367:1814–1820 [View Article] [PubMed]
    [Google Scholar]
  20. Zhu N, Zhang D, Wang W, Li X, Yang B et al. A novel coronavirus from patients with pneumonia in China, 2019. N Engl J Med 2020; 382:727–733 [View Article]
    [Google Scholar]
  21. Pandya MC, Callahan SM, Savchenko KG, Stobart CC. A contemporary view of respiratory syncytial virus (RSV) biology and strain-specific differences. Pathogens 2019; 8:67 [View Article] [PubMed]
    [Google Scholar]
  22. Schuster JE, Williams JV. Human metapneumovirus. Microbiol Spectr 2014; 2: [View Article]
    [Google Scholar]
  23. Li Y, Wang X, Blau DM, Caballero MT, Feikin DR et al. Global, regional, and national disease burden estimates of acute lower respiratory infections due to respiratory syncytial virus in children younger than 5 years in 2019: a systematic analysis. Lancet 2022; 399:2047–2064 [View Article] [PubMed]
    [Google Scholar]
  24. Lodi L, Catamerò F, Voarino M, Barbati F, Moriondo M et al. Epidemiology of respiratory syncytial virus in hospitalized children over a 9-year period and preventive strategy impact. Front Pharmacol 2024; 15:1381107 [View Article] [PubMed]
    [Google Scholar]
  25. Alzaydi M, Alosaimi A, Alghamdi AA, Bamogaddam IY, Altassan MA et al. Changes in seasonal respiratory viral infections among pediatric population around the COVID-19 pandemic; 2019–2023. Eur J Clin Microbiol Infect Dis 2024; 43:1589–1596 [View Article]
    [Google Scholar]
  26. Gombold JL, Hingley ST, Weiss SR. Fusion-defective mutants of mouse hepatitis virus A59 contain a mutation in the spike protein cleavage signal. J Virol 1993; 67:4504–4512 [View Article] [PubMed]
    [Google Scholar]
  27. Heminway BR, Yu Y, Tanaka Y, Perrine KG, Gustafson E et al. Analysis of respiratory syncytial virus F, G, and SH proteins in cell fusion. Virology 1994; 200:801–805 [View Article] [PubMed]
    [Google Scholar]
  28. Stobart CC, Lee AS, Lu X, Denison MR. Temperature-sensitive mutants and revertants in the coronavirus nonstructural protein 5 protease (3CLpro) define residues involved in long-distance communication and regulation of protease activity. J Virol 2012; 86:4801–4810 [View Article] [PubMed]
    [Google Scholar]
  29. DeFord DM, Nosek JM, Castiglia KR, Hasik EF, Franke ME et al. Evaluation of the role of respiratory syncytial virus surface glycoproteins F and G on viral stability and replication: implications for future vaccine design. J Gen Virol 2019; 100:1112–1122 [View Article] [PubMed]
    [Google Scholar]
  30. González-Parra G, Dobrovolny HM. The rate of viral transfer between upper and lower respiratory tracts determines RSV illness duration. J Math Biol 2019; 79:467–483 [View Article] [PubMed]
    [Google Scholar]
  31. González-Parra G, Dobrovolny HM. A quantitative assessment of dynamical differences of RSV infections in vitro and in vivo. Virology 2018; 523:129–139 [View Article] [PubMed]
    [Google Scholar]
  32. Nguyen VK, Binder SC, Boianelli A, Meyer-Hermann M, Hernandez-Vargas EA. Ebola virus infection modeling and identifiability problems. Front Microbiol 2015; 6:257 [View Article] [PubMed]
    [Google Scholar]
  33. Perelson AS, Neumann AU, Markowitz M, Leonard JM, Ho DD. HIV-1 dynamics in vivo: virion clearance rate, infected cell life-span, and viral generation time. Science 1996; 271:1582–1586 [View Article] [PubMed]
    [Google Scholar]
  34. Baccam P, Beauchemin C, Macken CA, Hayden FG, Perelson AS. Kinetics of influenza A virus infection in humans. J Virol 2006; 80:7590–7599 [View Article] [PubMed]
    [Google Scholar]
  35. Li P, Shan Y, Zheng W, Ou X, Mi D et al. Identification of H209 as essential for pH 8-triggered receptor-independent syncytium formation by S protein of mouse hepatitis virus A59. J Virol 2018; 92:e00209-18 [View Article] [PubMed]
    [Google Scholar]
  36. Heumann R, Duncan C, Stobart CC, Kaschner S. Dynamical differences in respiratory syncytial virus. Bull Math Biol 2021; 84:11 [View Article] [PubMed]
    [Google Scholar]
  37. Hotard AL, Shaikh FY, Lee S, Yan D, Teng MN et al. A stabilized respiratory syncytial virus reverse genetics system amenable to recombination-mediated mutagenesis. Virology 2012; 434:129–136 [View Article] [PubMed]
    [Google Scholar]
  38. Shcherbo D, Murphy CS, Ermakova GV, Solovieva EA, Chepurnykh TV et al. Far-red fluorescent tags for protein imaging in living tissues. Biochem J 2009; 418:567–574 [View Article] [PubMed]
    [Google Scholar]
  39. Rostad CA, Stobart CC, Gilbert BE, Pickles RJ, Hotard AL et al. A recombinant respiratory syncytial virus vaccine candidate attenuated by a low-fusion F protein is immunogenic and protective against challenge in cotton rats. J Virol 2016; 90:7508–7518 [View Article] [PubMed]
    [Google Scholar]
  40. Jessie B, Dobrovolny HM. The role of syncytia during viral infections. J Theor Biol 2021; 525:110749 [View Article] [PubMed]
    [Google Scholar]
  41. Holder BP, Beauchemin CAA. Exploring the effect of biological delays in kinetic models of influenza within a host or cell culture. BMC Public Health 2011; 11 Suppl 1:S10 [View Article] [PubMed]
    [Google Scholar]
  42. Beauchemin CAA, Miura T, Iwami S. Duration of SHIV production by infected cells is not exponentially distributed: implications for estimates of infection parameters and antiviral efficacy. Sci Rep 2017; 7:42765 [View Article] [PubMed]
    [Google Scholar]
  43. Knipe DM, Howley P. Fields Virology, 6th edition Lippincott Williams & Wilkins; 2013
    [Google Scholar]
  44. Currier MG, Lee S, Stobart CC, Hotard AL, Villenave R et al. EGFR interacts with the fusion protein of respiratory syncytial virus strain 2-20 and mediates infection and mucin expression. PLoS Pathog 2016; 12:e1005622 [View Article] [PubMed]
    [Google Scholar]
  45. Mehedi M, McCarty T, Martin SE, Le Nouën C, Buehler E et al. Actin-related protein 2 (ARP2) and virus-induced filopodia facilitate human respiratory syncytial virus spread. PLoS Pathog 2016; 12:e1006062 [View Article] [PubMed]
    [Google Scholar]
  46. Mehedi M, Collins PL, Buchholz UJ. A novel host factor for human respiratory syncytial virus. Commun Integr Biol 2017; 10:e1319025 [View Article] [PubMed]
    [Google Scholar]
  47. Krzyzaniak MA, Zumstein MT, Gerez JA, Picotti P, Helenius A. Host cell entry of respiratory syncytial virus involves macropinocytosis followed by proteolytic activation of the F protein. PLoS Pathog 2013; 9:e1003309 [View Article] [PubMed]
    [Google Scholar]
  48. Ulloa L, Serra R, Asenjo A, Villanueva N. Interactions between cellular actin and human respiratory syncytial virus (HRSV). Virus Res 1998; 53:13–25 [View Article] [PubMed]
    [Google Scholar]
  49. Burke E, Dupuy L, Wall C, Barik S. Role of cellular actin in the gene expression and morphogenesis of human respiratory syncytial virus. Virology 1998; 252:137–148 [View Article] [PubMed]
    [Google Scholar]
  50. Bartak M, Bąska P, Chodkowski M, Tymińska B, Bańbura MW et al. Neurons cytoskeletal architecture remodeling during the replication cycle of mouse coronavirus MHV-JHM: a morphological in vitro study. BMC Vet Res 2024; 20:18 [View Article] [PubMed]
    [Google Scholar]
  51. Zhao S, Gao J, Zhu L, Yang Q. Transmissible gastroenteritis virus and porcine epidemic diarrhoea virus infection induces dramatic changes in the tight junctions and microfilaments of polarized IPEC-J2 cells. Virus Res 2014; 192:34–45 [View Article] [PubMed]
    [Google Scholar]
  52. Mehedi M, Smelkinson M, Kabat J, Ganesan S, Collins PL et al. Multicolor stimulated emission depletion (STED) microscopy to generate high-resolution images of respiratory syncytial virus particles and infected cells. Bio Protoc 2017; 7:e2543 [View Article] [PubMed]
    [Google Scholar]
  53. Zhang Y, Zhang X, Li Z, Zhao W, Yang H et al. Single particle tracking reveals SARS-CoV-2 regulating and utilizing dynamic filopodia for viral invasion. Science Bulletin 2023; 68:2210–2224 [View Article]
    [Google Scholar]
  54. Freeman MC, Peek CT, Becker MM, Smith EC, Denison MR. Coronaviruses induce entry-independent, continuous macropinocytosis. mBio 2014; 5:e01340–14 [View Article] [PubMed]
    [Google Scholar]
  55. Zhang Y-Y, Liang R, Wang S-J, Ye Z-W, Wang T-Y et al. SARS-CoV-2 hijacks macropinocytosis to facilitate its entry and promote viral spike-mediated cell-to-cell fusion. J Biol Chem 2022; 298:102511 [View Article] [PubMed]
    [Google Scholar]
  56. Angelini MM, Akhlaghpour M, Neuman BW, Buchmeier MJ. Severe acute respiratory syndrome coronavirus nonstructural proteins 3, 4, and 6 induce double-membrane vesicles. mBio 2013; 4:mBio [View Article] [PubMed]
    [Google Scholar]
  57. Perlman S, Netland J. Coronaviruses post-SARS: update on replication and pathogenesis. Nat Rev Microbiol 2009; 7:439–450 [View Article] [PubMed]
    [Google Scholar]
  58. Saunders N, Monel B, Cayet N, Archetti L, Moreno H et al. Dynamic label-free analysis of SARS-CoV-2 infection reveals virus-induced subcellular remodeling. Nat Commun 2024; 15:4996 [View Article] [PubMed]
    [Google Scholar]
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