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Abstract

Human astroviruses (HAstVs) are small, non-enveloped icosahedral RNA viruses that are a significant cause of diarrhoea in young children. Despite their worldwide prevalence, HAstV pathogenesis studies and vaccine development remain challenging due to the lack of an animal model for HAstV infection. The recent development of a murine astrovirus (MuAstV) infection model in mice provides the opportunity to test proof-of-concept vaccines based on MuAstV antigens. To help establish a system in which an astrovirus capsid spike-based vaccine could be tested , we designed and produced a recombinant MuAstV capsid spike protein based on predicted secondary structure homology to HAstV spike proteins. The recombinant MuAstV spike can be expressed with high efficiency in and retains antigenicity to polyclonal antibodies elicited by MuAstV infection. We determined the crystal structure of the MuAstV spike to 1.75 Å and assessed its structural conservation with HAstV capsid spike. Despite low sequence identity between the MuAstV and HAstV spikes and differences in their overall shapes, they share related structural folds. Additionally, we found that vaccination with MuAstV spike induced anti-MuAstV-spike antibodies, highlighting that the recombinant spike is immunogenic. These studies lay a foundation for future MuAstV challenge studies to test whether MuAstV spike can be the basis of an effective vaccine.

Funding
This study was supported by the:
  • National Institute of General Medical Sciences (Award T32 GM133391)
    • Principle Award Recipient: NataliePedicino
  • National Institute of General Medical Sciences (Award T32 GM133391)
    • Principle Award Recipient: SarahLanning
  • Division of Intramural Research, National Institute of Allergy and Infectious Diseases (Award K22 AI156116)
    • Principle Award Recipient: ValerieCortez
  • Division of Intramural Research, National Institute of Allergy and Infectious Diseases (Award R01 AI144090)
    • Principle Award Recipient: RebeccaM. DuBois
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/content/journal/jgv/10.1099/jgv.0.001913
2023-11-01
2024-06-20
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References

  1. Madeley CR, Cosgrove BP. Viruses in infantile gastroenteritis. Lancet 1975; 2:124 [View Article] [PubMed]
    [Google Scholar]
  2. Arias CF, DuBois RM. The astrovirus capsid: a review. Viruses 2017; 9:15 [View Article] [PubMed]
    [Google Scholar]
  3. Olortegui MP, Rouhani S, Yori PP, Salas MS, Trigoso DR et al. Astrovirus infection and diarrhea in 8 countries. Pediatrics 2018; 141:e20171326 [View Article] [PubMed]
    [Google Scholar]
  4. Gaggero A, O’Ryan M, Noel JS, Glass RI, Monroe SS et al. Prevalence of astrovirus infection among Chilean children with acute gastroenteritis. J Clin Microbiol 1998; 36:3691–3693 [View Article] [PubMed]
    [Google Scholar]
  5. Palombo EA, Bishop RF. Annual incidence, serotype distribution, and genetic diversity of human astrovirus isolates from hospitalized children in Melbourne, Australia. J Clin Microbiol 1996; 34:1750–1753 [View Article] [PubMed]
    [Google Scholar]
  6. Shastri S, Doane AM, Gonzales J, Upadhyayula U, Bass DM. Prevalence of astroviruses in a children’s hospital. J Clin Microbiol 1998; 36:2571–2574 [View Article] [PubMed]
    [Google Scholar]
  7. Meyer L, Delgado-Cunningham K, Lorig-Roach N, Ford J, DuBois RM. Human astrovirus 1-8 seroprevalence evaluation in a United States adult population. Viruses 2021; 13:979 [View Article] [PubMed]
    [Google Scholar]
  8. Koopmans MPG, Bijen MHL, Monroe SS, Vinjé J. Age-stratified seroprevalence of neutralizing antibodies to astrovirus types 1 to 7 in humans in The Netherlands. Clin Diagn Lab Immunol 1998; 5:33–37 [View Article]
    [Google Scholar]
  9. Kurtz J, Lee T. Astrovirus gastroenteritis age distribution of antibody. Med Microbiol Immunol 1978; 166:227–230 [View Article] [PubMed]
    [Google Scholar]
  10. Kurtz JB, Lee TW, Craig JW, Reed SE. Astrovirus infection in volunteers. J Med Virol 1979; 3:221–230 [View Article] [PubMed]
    [Google Scholar]
  11. Quan PL, Wagner TA, Briese T, Torgerson TR, Hornig M et al. Astrovirus encephalitis in boy with X-linked agammaglobulinemia. Emerg Infect Dis 2010; 16:918–925 [View Article] [PubMed]
    [Google Scholar]
  12. Brown JR, Morfopoulou S, Hubb J, Emmett WA, Ip W et al. Astrovirus VA1/HMO-C: an increasingly recognized neurotropic pathogen in immunocompromised patients. Clin Infect Dis 2015; 60:881–888 [View Article]
    [Google Scholar]
  13. Cordey S, Vu D-L, Schibler M, L’Huillier AG, Brito F et al. Astrovirus MLB2, a new gastroenteric virus associated with meningitis and disseminated infection. Emerg Infect Dis 2016; 22:846–853 [View Article] [PubMed]
    [Google Scholar]
  14. Koci MD, Moser LA, Kelley LA, Larsen D, Brown CC et al. Astrovirus induces diarrhea in the absence of inflammation and cell death. J Virol 2003; 77:11798–11808 [View Article] [PubMed]
    [Google Scholar]
  15. Koci MD, Kelley LA, Larsen D, Schultz-Cherry S. Astrovirus-induced synthesis of nitric oxide contributes to virus control during infection. J Virol 2004; 78:1564–1574 [View Article] [PubMed]
    [Google Scholar]
  16. Cortez V, Meliopoulos VA, Karlsson EA, Hargest V, Johnson C et al. Astrovirus biology and pathogenesis. Annu Rev Virol 2017; 4:327–348 [View Article] [PubMed]
    [Google Scholar]
  17. DuBois RM, Freiden P, Marvin S, Reddivari M, Heath RJ et al. Crystal structure of the avian astrovirus capsid spike. J Virol 2013; 87:7853–7863 [View Article] [PubMed]
    [Google Scholar]
  18. Kjeldsberg E, Hem A. Detection of astroviruses in gut contents of nude and normal mice. Brief report. Arch Virol 1985; 84:135–140 [View Article] [PubMed]
    [Google Scholar]
  19. Ricart Arbona RJ, Kelly S, Wang C, Dhawan RK, Henderson KS et al. Serendipitous discovery of a novel murine astrovirus contaminating a murine helper T-cell line and incapable of infecting highly immunodeficient mice. comp med 2020; 70:359–369 [View Article]
    [Google Scholar]
  20. Yokoyama CC, Loh J, Zhao G, Stappenbeck TS, Wang D et al. Adaptive immunity restricts replication of novel murine astroviruses. J Virol 2012; 86:12262–12270 [View Article]
    [Google Scholar]
  21. Compton SR, Booth CJ, Macy JD. Murine astrovirus infection and transmission in neonatal CD1 mice. J Am Assoc Lab Anim Sci 2017; 56:402–411 [PubMed]
    [Google Scholar]
  22. Sebire NJ, Malone M, Shah N, Anderson G, Gaspar HB et al. Pathology of astrovirus associated diarrhoea in a paediatric bone marrow transplant recipient. J Clin Pathol 2004; 57:1001–1003 [View Article] [PubMed]
    [Google Scholar]
  23. Cortez V, Sharp B, Yao J, Livingston B, Vogel P et al. Characterizing a murine model for astrovirus using viral isolates from persistently infected immunocompromised mice. J Virol 2019; 93:e00223-19 [View Article] [PubMed]
    [Google Scholar]
  24. Cortez V, Boyd DF, Crawford JC, Sharp B, Livingston B et al. Astrovirus infects actively secreting goblet cells and alters the gut mucus barrier. Nat Commun 2020; 11:2097 [View Article] [PubMed]
    [Google Scholar]
  25. Ingle H, Hassan E, Gawron J, Mihi B, Li Y et al. Murine astrovirus tropism for goblet cells and enterocytes facilitates an IFN-λ response in vivo and in enteroid cultures. Mucosal Immunol 2021; 14:751–761 [View Article] [PubMed]
    [Google Scholar]
  26. Triana S, Stanifer ML, Metz-Zumaran C, Shahraz M, Mukenhirn M et al. Single-cell transcriptomics reveals immune response of intestinal cell types to viral infection. Mol Syst Biol 2021; 17:e9833 [View Article] [PubMed]
    [Google Scholar]
  27. Kolawole AO, Mirabelli C, Hill DR, Svoboda SA, Janowski AB et al. Astrovirus replication in human intestinal enteroids reveals multi-cellular tropism and an intricate host innate immune landscape. PLoS Pathog 2019; 15:e1008057 [View Article] [PubMed]
    [Google Scholar]
  28. Banos-Lara M del R, Méndez E. Role of individual caspases induced by astrovirus on the processing of its structural protein and its release from the cell through a non-lytic mechanism. Virology 2010; 401:322–332 [View Article] [PubMed]
    [Google Scholar]
  29. Dryden KA, Tihova M, Nowotny N, Matsui SM, Mendez E et al. Immature and mature human astrovirus: structure, conformational changes, and similarities to hepatitis E virus. J Mol Biol 2012; 422:650–658 [View Article] [PubMed]
    [Google Scholar]
  30. Ricemeyer L, Aguilar-Hernández N, López T, Espinosa R, Lanning S et al. Structures of two human astrovirus capsid/neutralizing antibody complexes reveal distinct epitopes and inhibition of virus attachment to cells. J Virol 2022; 96:e0141521 [View Article] [PubMed]
    [Google Scholar]
  31. Espinosa R, López T, Bogdanoff WA, Espinoza MA, López S et al. Isolation of neutralizing monoclonal antibodies to human astrovirus and characterization of virus variants that escape neutralization. J Virol 2019; 93:jvi [View Article]
    [Google Scholar]
  32. Bass DM, Upadhyayula U. Characterization of human serotype 1 astrovirus-neutralizing epitopes. J Virol 1997; 71:8666–8671 [View Article] [PubMed]
    [Google Scholar]
  33. Sanchez-Fauquier A, Carrascosa AL, Carrascosa JL, Otero A, Glass RI et al. Characterization of a human astrovirus serotype 2 structural protein (VP26) that contains an epitope involved in virus neutralization. Virology 1994; 201:312–320 [View Article] [PubMed]
    [Google Scholar]
  34. Zimmermann L, Stephens A, Nam S-Z, Rau D, Kübler J et al. A completely reimplemented MPI bioinformatics toolkit with a new HHpred server at its core. J Mol Biol 2018; 430:2237–2243 [View Article] [PubMed]
    [Google Scholar]
  35. Emsley P, Lohkamp B, Scott WG, Cowtan K. Features and development of Coot. Acta Crystallogr D Biol Crystallogr 2010; 66:486–501 [View Article] [PubMed]
    [Google Scholar]
  36. Liebschner D, Afonine PV, Baker ML, Bunkóczi G, Chen VB et al. Macromolecular structure determination using X-rays, neutrons and electrons: recent developments in Phenix. Acta Crystallogr D Struct Biol 2019; 75:861–877 [View Article]
    [Google Scholar]
  37. Edgar RC. MUSCLE: multiple sequence alignment with high accuracy and high throughput. Nucleic Acids Res 2004; 32:1792–1797 [View Article] [PubMed]
    [Google Scholar]
  38. Jones DT, Taylor WR, Thornton JM. The rapid generation of mutation data matrices from protein sequences. Bioinformatics 1992; 8:275–282 [View Article]
    [Google Scholar]
  39. Kumar S, Stecher G, Li M, Knyaz C, Tamura K. MEGA X: Molecular Evolutionary Genetics Analysis across computing platforms. Mol Biol Evol 2018; 35:1547–1549 [View Article] [PubMed]
    [Google Scholar]
  40. Dong J, Dong L, Méndez E, Tao Y. Crystal structure of the human astrovirus capsid spike. Proc Natl Acad Sci U S A 2011; 108:12681–12686 [View Article] [PubMed]
    [Google Scholar]
  41. Zheng H, Cooper DR, Porebski PJ, Shabalin IG, Handing KB et al. CheckMyMetal: a macromolecular metal-binding validation tool. Acta Crystallogr D Struct Biol 2017; 73:223–233 [View Article] [PubMed]
    [Google Scholar]
  42. Zhang Y, Skolnick J. TM-align: a protein structure alignment algorithm based on the TM-score. Nucleic Acids Res 2005; 33:2302–2309 [View Article] [PubMed]
    [Google Scholar]
  43. Delgado-Cunningham K, López T, Khatib F, Arias CF, DuBois RM. Structure of the divergent human astrovirus MLB capsid spike. Structure 2022; 30:1573–1581 [View Article] [PubMed]
    [Google Scholar]
  44. Schmidt K, Butt J, Mauter P, Vogel K, Erles-Kemna A et al. Development of a multiplex serological assay reveals a worldwide distribution of murine astrovirus infections in laboratory mice. PLoS ONE 2017; 12:e0187174 [View Article]
    [Google Scholar]
  45. Kawagishi T, Sánchez-Tacuba L, Feng N, Costantini VP, Tan M et al. Mucosal and systemic neutralizing antibodies to norovirus induced in infant mice orally inoculated with recombinant rotaviruses. Proc Natl Acad Sci U S A 2023; 120:e2214421120 [View Article] [PubMed]
    [Google Scholar]
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