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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 1

Human norovirus–positive fecal samples tested on 80 jejunal HIEs in study of human norovirus replication in HIEs

Genotype* No. samples
Mean norovirus RNA copies/μL
Patient age group Collection date†
Storage condition‡
Outbreak or sporadic
No. inactivated/ no. samples %
Capsid
RdRp
Years
No.
GI.1
GI.P1
4
0.5–19.3 × 103
>18
4
2000
−70°C
Sporadic
0/4 (0)
GI.3
GI.P3 2 0.3–59.4 × 104 0–12 1 2013 Jul −70°C Sporadic 0/4 (0)
>18 1 2005 −70°C Outbreak
GI.Pd
2
1.2–8.7 × 104
0–12
1 2015 Nov −70°C Sporadic
1
2017 Feb
−70°C
Sporadic
GI.4
GI.P4
1
2.2 × 102
>18
1
2000
−70°C
Outbreak
0/1 (0)
GI.7
GI.P7
3
2.6–16.4 × 103
0–12
1 2010 Nov −70°C Sporadic 0/3 (0)
1 2014 Mar −70°C Sporadic
1
2016 Dec
−70°C
Sporadic
GII.1
GII.Pg
1
1.4 × 105
0–12
1
2017 May
−70°C
Sporadic
1/1 (100)
GII.2
GII.P16
3
0.2–52.6 × 103
0–12
2 2017 Feb −70°C Sporadic 1/2 (50)
1
2017 Mar
−70°C
Sporadic
0/1 (0)
GII.3
GII.P21 2 1.0–6.4 × 105 0–12 1 2012 Mar −70°C Sporadic 0/1 (0)
1 2015 May −70°C Sporadic
GII.P12 2 1.6–4.4 × 106 0–12 2 2012 Aug −70°C Sporadic 1/2 (50)
GII.P16
4
0.2–141.1 × 104
0–12
1 2012 Mar −70°C Sporadic 0/4 (0)
1 2012 Jun −70°C Sporadic
1 2015 Oct −70°C Sporadic
1
2016 Dec
−70°C
Sporadic
GII.4 Den Haag
GII.P4 Den Haag
3
1.3–161.6 × 104
>18 1 2010 May 4°C Outbreak 0/1 (0)
0–12
1 2013 Jun −70°C Sporadic 0/1 (0)
1
2015 May
−70°C
Sporadic
1/1 (100)
GII.4 New Orleans
GII.P4 New Orleans
1
4.1 × 105
0–12
1
2013 Apr
−70°C
Sporadic
0/1 (0)
GII.4 Sydney
GII.Pe 22 3.5 × 103–2.1 × 107 0–12 3 2012 −70°C Sporadic 1/3 (30)
1 2015 Jan −70°C Sporadic 1/1 (100)
7 2015 Feb −70°C Sporadic 3/7 (40)
1 2015 Mar −70°C Sporadic 1/1 (100)
1 2015 Apr −70°C Sporadic 1/1 (100)
1 2015 Oct −70°C Sporadic 0/1 (0)
1 2016 Apr −70°C Sporadic 0/1 (0)
>18 1 2011 −70°C Outbreak 0/1 (0)
1 2013 −70°C Outbreak 0/1 (0)
1 2012 May 4°C Outbreak 0/1 (0)
2 2016 Apr 4°C Outbreak 1/2 (50)
2 2016 May 4°C Outbreak 0/2 (0)
GII.P16 13 0.4 × 103–6.1 × 106 0–12 2 2016 Dec −70°C Sporadic 1/2 (50)
6 2017 Feb −70°C Sporadic 0/6 (0)
5 2017 Mar −70°C Sporadic 0/5 (0)
GII.P4 New Orleans
4
0.6–14.9 × 104
0–12
1 2015 Jan −70°C Sporadic 0/4 (0)
1 2017 Jan −70°C Sporadic
1 2017 Feb −70°C Sporadic
1
2017 Mar
−70°C
Sporadic
GII.5
GII.P22
1
1.1 × 104
0–12
1
2010 Nov
−70°C
Sporadic
0/1 (0)
GII.6B
GII.P7
1
7.6 × 104
0–12
1
2015 Jan
−70°C
Sporadic
0/1 (0)
GII.6
GII.P7
4
0.1–8.4 × 106
0–12
1 2012 Dec −70°C Sporadic 0/4 (0)
2 2015 Jan −70°C Sporadic
1
2017 Jan
−70°C
Sporadic
GII.7
GII.P7
2
0.3–9.1 × 106
>18 1 2010 Oct −70°C Sporadic 0/2 (0)
0–12
1
2012 Aug
−70°C
Sporadic
GII.14
GII.P7
1
6.1 × 104
0–12
1
2016 Dec
−70°C
Sporadic
1/1 (100)
GII.17
GII.Pe
1
3.4 × 105
0–12
1
2010 Oct
−70°C
Sporadic
1/1 (100)
GIV
GIV.P1
3
0.3–13.6 × 103
>18
3
2016 May
4°C
Outbreak
0/3 (0)
Total 80 80 16/80 (20)

*Dual genotyping based on sequencing partial RdRp and capsid regions (26,27). HIE, human intestinal enteroids; RdRp, RNA-dependent RNA polymerase.
†When month was not available, only year of collection is reported.
‡Samples were stored at the indicated temperature.

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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;14:72530.DOIPubMed
  2. Duizer  E, Schwab  KJ, Neill  FH, Atmar  RL, Koopmans  MP, Estes  MK. Laboratory efforts to cultivate noroviruses. J Gen Virol. 2004;85:7987.DOIPubMed
  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;13:396403.DOIPubMed
  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;19:4318.DOIPubMed
  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.DOIPubMed
  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;159:25766.DOIPubMed
  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.DOIPubMed
  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;406:111.DOIPubMed
  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;346:7559.DOIPubMed
  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;10:193947.DOIPubMed
  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;97:2291300.DOIPubMed
  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;141:176272.DOIPubMed
  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;90:4356.DOIPubMed
  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;64:91120.DOIPubMed
  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;291:375966.DOIPubMed
  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;353:138793.DOIPubMed
  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.DOIPubMed
  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;77:547682.DOIPubMed
  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;50:2614.PubMed
  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;76:100616.DOIPubMed
  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;8:112.PubMed
  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;78:2249.DOIPubMed
  23. Bozkurt  H, D’Souza  DH, Davidson  PM. Thermal inactivation of foodborne enteric viruses and their viral surrogates in foods. J Food Prot. 2015;78:1597617.DOIPubMed
  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;80:574351.DOIPubMed
  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;69:27615.DOIPubMed
  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;55:220821.DOIPubMed
  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;52:14755.DOIPubMed
  28. Reed  LJ, Muench  H. A simple method of estimating fifty per cent endpoints. Am J Hyg. 1938;27:49397.
  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;114:16727.DOIPubMed
  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:E5709.DOIPubMed
  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.DOIPubMed
  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;11:10612.DOIPubMed

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Page updated: July 18, 2018
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