Skip directly to site content Skip directly to page options Skip directly to A-Z link Skip directly to A-Z link Skip directly to A-Z link
Volume 24, Number 8—August 2018
Research

Human Norovirus Replication in Human Intestinal Enteroids as Model to Evaluate Virus Inactivation

Veronica CostantiniComments to Author , Esther K. Morantz, Hannah Browne, Khalil Ettayebi, Xi-Lei Zeng, Robert L. Atmar, Mary K. Estes, and Jan Vinjé
Author affiliations: Centers for Disease Control and Prevention, Atlanta, Georgia, USA (V. Costantini, J. Vinjé); Synergy America, Inc., Atlanta (E.K. Morantz); Oak Ridge Institute for Science and Education, Oak Ridge, Tennessee, USA (H. Browne); Baylor College of Medicine, Houston, Texas, USA (K. Ettayebi, X.-L. Zeng, R.L. Atmar, M.K. Estes)

Main Article

Table 2

Norovirus fecal filtrates successfully cultivated in study of human norovirus replication in human intestinal enteroids

Sample ID Genotype, RdRp-Capsid* Mean norovirus RNA copies/μL Participant age, mo Collection date† Storage condition‡ Fold virus RNA increase, log10§
NP01015 GII.Pg-GII.1 1.4 × 105 4 2017 Mar −70°C 696 (2.8)
NP36980 GII.P16-GII.2 1.9 × 103 132 2017 Feb −70°C 56 (1.8)
N741656 GII.P12-GII.3 1.4 × 105 9 2012 Aug −70°C 181 (2.3)
G3868 GII.P4 Den Haag-GII.4 Den Haag 2.0 × 105 12 2015 May −70°C 155 (2.2)
G3829 GII.P4 New Orleans-GII.4 New Orleans 4.1 × 105 5 2013 Apr −70°C 227 (2.4)
A5413 GII.Pe-GII.4 Sydney 1.6 × 106 12 2012 −70°C 487 (2.7)
R3702 GII.Pe-GII.4 Sydney 7.7 × 106 46 2015 Jan −70°C 1236 (3.0)
V0882 GII.Pe-GII.4 Sydney 4.0 × 106 20 2015 Feb −70°C 998 (3.0)
V0900 GII.Pe-GII.4 Sydney 5.1 × 105 17 2015 Feb −70°C 1102 (3.0)
H4423 GII.Pe-GII.4 Sydney 5.9 × 105 12 2015 Feb −70°C 125 (2.1)
H4587 GII.Pe-GII.4 Sydney 4.4 × 106 21 2015 Apr −70°C 448 (2.7)
C8993 GII.Pe-GII.4 Sydney 1.7 × 107 19 2015 Mar −70°C 223 (2.3)
CDC830¶ GII.Pe-GII.4 Sydney 2.9 × 106 996 2016 Apr 4°C 121 (2.1)
NP36995 GII.P16-GII.4 Sydney 2.1 × 106 14 2016 Dec −70°C 340 (2.5)
NP36994 GII.P7-GII.14 6.1 × 104 22 2016 Dec −70°C 47 (1.6)
G3734 GII.Pe-GII.17 3.4 × 105 11 2010 Oct −70°C 96 (1.9)

*Dual genotyping based on sequencing partial RdRp and capsid regions (26,27). RdRp, RNA-dependent RNA polymerase.
†When month was not available, only year of collection is reported.
‡Samples were kept from collection time until testing at the indicated temperature.
§Virus RNA increase after 3 days postinfection expressed as fold increase and log10 fold increase. For each fecal filtrate, data represent mean viral RNA increase from 3 experiments with 3 technical replicates each.
¶CDC830 was stored at 4°C for 2 mo before cultivation.

Main Article

References
  1. Ahmed  SM, Hall  AJ, Robinson  AE, Verhoef  L, Premkumar  P, Parashar  UD, et al. Global prevalence of norovirus in cases of gastroenteritis: a systematic review and meta-analysis. Lancet Infect Dis. 2014;14:725–30.DOIPubMedGoogle Scholar
  2. Duizer  E, Schwab  KJ, Neill  FH, Atmar  RL, Koopmans  MP, Estes  MK. Laboratory efforts to cultivate noroviruses. J Gen Virol. 2004;85:79–87.DOIPubMedGoogle Scholar
  3. Straub  TM, Höner zu Bentrup  K, Orosz-Coghlan  P, Dohnalkova  A, Mayer  BK, Bartholomew  RA, et al. In vitro cell culture infectivity assay for human noroviruses. Emerg Infect Dis. 2007;13:396–403.DOIPubMedGoogle Scholar
  4. Herbst-Kralovetz  MM, Radtke  AL, Lay  MK, Hjelm  BE, Bolick  AN, Sarker  SS, et al. Lack of norovirus replication and histo-blood group antigen expression in 3-dimensional intestinal epithelial cells. Emerg Infect Dis. 2013;19:431–8.DOIPubMedGoogle Scholar
  5. Papafragkou  E, Hewitt  J, Park  GW, Greening  G, Vinjé  J. Challenges of culturing human norovirus in three-dimensional organoid intestinal cell culture models. PLoS One. 2013;8:e63485.DOIPubMedGoogle Scholar
  6. Takanashi  S, Saif  LJ, Hughes  JH, Meulia  T, Jung  K, Scheuer  KA, et al. Failure of propagation of human norovirus in intestinal epithelial cells with microvilli grown in three-dimensional cultures. Arch Virol. 2014;159:257–66.DOIPubMedGoogle Scholar
  7. Wobus  CE, Karst  SM, Thackray  LB, Chang  KO, Sosnovtsev  SV, Belliot  G, et al. Replication of Norovirus in cell culture reveals a tropism for dendritic cells and macrophages. PLoS Biol. 2004;2:e432.DOIPubMedGoogle Scholar
  8. Lay  MK, Atmar  RL, Guix  S, Bharadwaj  U, He  H, Neill  FH, et al. Norwalk virus does not replicate in human macrophages or dendritic cells derived from the peripheral blood of susceptible humans. Virology. 2010;406:1–11.DOIPubMedGoogle Scholar
  9. Jones  MK, Watanabe  M, Zhu  S, Graves  CL, Keyes  LR, Grau  KR, et al. Enteric bacteria promote human and mouse norovirus infection of B cells. Science. 2014;346:755–9.DOIPubMedGoogle Scholar
  10. Jones  MK, Grau  KR, Costantini  V, Kolawole  AO, de Graaf  M, Freiden  P, et al. Human norovirus culture in B cells. Nat Protoc. 2015;10:1939–47.DOIPubMedGoogle Scholar
  11. Karandikar  UC, Crawford  SE, Ajami  NJ, Murakami  K, Kou  B, Ettayebi  K, et al. Detection of human norovirus in intestinal biopsies from immunocompromised transplant patients. J Gen Virol. 2016;97:2291–300.DOIPubMedGoogle Scholar
  12. Sato  T, Stange  DE, Ferrante  M, Vries  RG, Van Es  JH, Van den Brink  S, et al. Long-term expansion of epithelial organoids from human colon, adenoma, adenocarcinoma, and Barrett’s epithelium. Gastroenterology. 2011;141:1762–72.DOIPubMedGoogle Scholar
  13. Saxena  K, Blutt  SE, Ettayebi  K, Zeng  XL, Broughman  JR, Crawford  SE, et al. Human intestinal enteroids: a new model to study human rotavirus infection, host restriction, and pathophysiology. J Virol. 2015;90:43–56.DOIPubMedGoogle Scholar
  14. VanDussen  KL, Marinshaw  JM, Shaikh  N, Miyoshi  H, Moon  C, Tarr  PI, et al. Development of an enhanced human gastrointestinal epithelial culture system to facilitate patient-based assays. Gut. 2015;64:911–20.DOIPubMedGoogle Scholar
  15. Zachos  NC, Kovbasnjuk  O, Foulke-Abel  J, In  J, Blutt  SE, de Jonge  HR, et al. Human enteroids/colonoids and intestinal organoids functionally recapitulate normal intestinal physiology and pathophysiology. J Biol Chem. 2016;291:3759–66.DOIPubMedGoogle Scholar
  16. Ettayebi  K, Crawford  SE, Murakami  K, Broughman  JR, Karandikar  U, Tenge  VR, et al. Replication of human noroviruses in stem cell-derived human enteroids. Science. 2016;353:1387–93.DOIPubMedGoogle Scholar
  17. Zou  WY, Blutt  SE, Crawford  SE, Ettayebi  K, Zeng  XL, Saxena  K, et al. Human intestinal enteroids: new models to study gastrointestinal virus infections. Methods Mol Biol. 2017.DOIPubMedGoogle Scholar
  18. Leon  JS, Kingsley  DH, Montes  JS, Richards  GP, Lyon  GM, Abdulhafid  GM, et al. Randomized, double-blinded clinical trial for human norovirus inactivation in oysters by high hydrostatic pressure processing. Appl Environ Microbiol. 2011;77:5476–82.DOIPubMedGoogle Scholar
  19. Keswick  BH, Satterwhite  TK, Johnson  PC, DuPont  HL, Secor  SL, Bitsura  JA, et al. Inactivation of Norwalk virus in drinking water by chlorine. Appl Environ Microbiol. 1985;50:261–4.PubMedGoogle Scholar
  20. Hoelzer  K, Fanaselle  W, Pouillot  R, Van Doren  JM, Dennis  S. Virus inactivation on hard surfaces or in suspension by chemical disinfectants: systematic review and meta-analysis of norovirus surrogates. J Food Prot. 2013;76:1006–16.DOIPubMedGoogle Scholar
  21. Zonta  W, Mauroy  A, Farnir  F, Thiry  E. Comparative virucidal efficacy of seven disinfectants against murine norovirus and feline calicivirus, surrogates of human norovirus. Food Environ Virol. 2016;8:1–12.PubMedGoogle Scholar
  22. Mormann  S, Heißenberg  C, Pfannebecker  J, Becker  B. Tenacity of human norovirus and the surrogates feline calicivirus and murine norovirus during long-term storage on common nonporous food contact surfaces. J Food Prot. 2015;78:224–9.DOIPubMedGoogle Scholar
  23. Bozkurt  H, D’Souza  DH, Davidson  PM. Thermal inactivation of foodborne enteric viruses and their viral surrogates in foods. J Food Prot. 2015;78:1597–617.DOIPubMedGoogle Scholar
  24. Cromeans  T, Park  GW, Costantini  V, Lee  D, Wang  Q, Farkas  T, et al. Comprehensive comparison of cultivable norovirus surrogates in response to different inactivation and disinfection treatments. Appl Environ Microbiol. 2014;80:5743–51.DOIPubMedGoogle Scholar
  25. Cannon  JL, Papafragkou  E, Park  GW, Osborne  J, Jaykus  LA, Vinjé  J. Surrogates for the study of norovirus stability and inactivation in the environment: aA comparison of murine norovirus and feline calicivirus. J Food Prot. 2006;69:2761–5.DOIPubMedGoogle Scholar
  26. Cannon  JL, Barclay  L, Collins  NR, Wikswo  ME, Castro  CJ, Magaña  LC, et al. Genetic and epidemiologic trends of norovirus outbreaks in the United States from 2013 to 2016 demonstrated emergence of novel GII.4 recombinant viruses. J Clin Microbiol. 2017;55:2208–21.DOIPubMedGoogle Scholar
  27. Vega  E, Barclay  L, Gregoricus  N, Shirley  SH, Lee  D, Vinjé  J. Genotypic and epidemiologic trends of norovirus outbreaks in the United States, 2009 to 2013. J Clin Microbiol. 2014;52:147–55.DOIPubMedGoogle Scholar
  28. Reed  LJ, Muench  H. A simple method of estimating fifty per cent endpoints. Am J Hyg. 1938;27:493–97.
  29. Drummond  CG, Bolock  AM, Ma  C, Luke  CJ, Good  M, Coyne  CB. Enteroviruses infect human enteroids and induce antiviral signaling in a cell lineage-specific manner. Proc Natl Acad Sci U S A. 2017;114:1672–7.DOIPubMedGoogle Scholar
  30. Saxena  K, Simon  LM, Zeng  XL, Blutt  SE, Crawford  SE, Sastri  NP, et al. A paradox of transcriptional and functional innate interferon responses of human intestinal enteroids to enteric virus infection. Proc Natl Acad Sci U S A. 2017;114:E570–9.DOIPubMedGoogle Scholar
  31. Kingsley  DH, Fay  JP, Calci  K, Pouillot  R, Woods  J, Chen  H, et al. Evaluation of chlorine treatment levels for inactivation of human norovirus and MS2 bacteriophage during sewage treatment. Appl Environ Microbiol. 2017;83:e01270-17.DOIPubMedGoogle Scholar
  32. Yin  X, Farin  HF, van Es  JH, Clevers  H, Langer  R, Karp  JM. Niche-independent high-purity cultures of Lgr5+ intestinal stem cells and their progeny. Nat Methods. 2014;11:106–12.DOIPubMedGoogle Scholar

Main Article

Page created: July 18, 2018
Page updated: July 18, 2018
Page reviewed: July 18, 2018
The conclusions, findings, and opinions expressed by authors contributing to this journal do not necessarily reflect the official position of the U.S. Department of Health and Human Services, the Public Health Service, the Centers for Disease Control and Prevention, or the authors' affiliated institutions. Use of trade names is for identification only and does not imply endorsement by any of the groups named above.
file_external