Journal of Veterinary Science

  • 제24권4호
  • /
  • Pages.53.1-53.12
  • /
  • 2023
  • /
  • 1229-845X(pISSN)
  • /
  • 1976-555X(eISSN)
대한수의학회 (The Korean Society of Veterinary Science)
권호 표지
DOI QR코드

DOI QR Code

Evaluation of porcine intestinal organoids as an in vitro model for mammalian orthoreovirus 3 infection

  • Se-A Lee (Viral Disease Division, Animal and Plant Quarantine Agency) ;
  • Hye Jeong Lee (Viral Disease Division, Animal and Plant Quarantine Agency) ;
  • Na-Yeon Gu (Viral Disease Division, Animal and Plant Quarantine Agency) ;
  • Yu-Ri Park (Viral Disease Division, Animal and Plant Quarantine Agency) ;
  • Eun-Ju Kim (Viral Disease Division, Animal and Plant Quarantine Agency) ;
  • Seok-Jin Kang (Viral Disease Division, Animal and Plant Quarantine Agency) ;
  • Bang-Hun Hyun (Viral Disease Division, Animal and Plant Quarantine Agency) ;
  • Dong-Kun Yang (Viral Disease Division, Animal and Plant Quarantine Agency)
  • 투고 : 2023.01.16
  • 심사 : 2023.06.16
  • 발행 : 2023.07.31
PDF 다운로드
⟨ 이전 논문 다음 논문 ⟩

초록

Background: Mammalian orthoreovirus type 3 (MRV3), which is responsible for gastroenteritis in many mammalian species including pigs, has been isolated from piglets with severe diarrhea. However, the use of pig-derived cells as an infection model for swine-MRV3 has rarely been studied. Objectives: This study aims to establish porcine intestinal organoids (PIOs) and examine their susceptibility as an in vitro model for intestinal MRV3 infection. Methods: PIOs were isolated and established from the jejunum of a miniature pig. Established PIOs were characterized using polymerase chain reaction (PCR) and immunofluorescence assays (IFAs) to confirm the expression of small intestine-specific genes and proteins, such as Lgr5, LYZI, Mucin-2, ChgA, and Villin. The monolayered PIOs and three-dimensional (3D) PIOs, obtained through their distribution to expose the apical surface, were infected with MRV3 for 2 h, washed with Dulbecco's phosphate-buffered saline, and observed. Viral infection was confirmed using PCR and IFA. We performed quantitative real-time reverse transcription-PCR to assess changes in viral copy numbers and gene expressions linked to intestinal epithelial genes and antiviral activity. Results: The established PIOs have molecular characteristics of intestinal organoids. Infected PIOs showed delayed proliferation with disruption of structures. In addition, infection with MRV3 altered the gene expression linked to intestinal epithelial cells and antiviral activity, and these effects were observed in both 2D and 3D models. Furthermore, viral copy numbers in the supernatant of both models increased in a time-dependent manner. Conclusions: We suggest that PIOs can be an in vitro model to study the infection mechanism of MRV3 in detail, facilitating pharmaceutical development.

키워드

과제정보

This study was supported financially by a grant (N-1543083-2023-32-01) from the Animal and Plant Quarantine Agency, Ministry of Agriculture, Food and Rural Affairs (MAFRA), Korea.

참고문헌

  1. Sato T, Vries RG, Snippert HJ, van de Wetering M, Barker N, Stange DE, et al. Single Lgr5 stem cells build crypt-villus structures in vitro without a mesenchymal niche. Nature. 2009;459(7244):262-265. https://doi.org/10.1038/nature07935
  2. 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(6306):1387-1393. https://doi.org/10.1126/science.aaf5211
  3. Gonzalez LM, Williamson I, Piedrahita JA, Blikslager AT, Magness ST. Cell lineage identification and stem cell culture in a porcine model for the study of intestinal epithelial regeneration. PLoS One. 2013;8(6):e66465.
  4. Derricott H, Luu L, Fong WY, Hartley CS, Johnston LJ, Armstrong SD, et al. Developing a 3D intestinal epithelium model for livestock species. Cell Tissue Res. 2019;375(2):409-424. https://doi.org/10.1007/s00441-018-2924-9
  5. Powell RH, Behnke MS. WRN conditioned media is sufficient for in vitro propagation of intestinal organoids from large farm and small companion animals. Biol Open. 2017;6(5):698-705.
  6. Acharya M, Arsi K, Donoghue AM, Liyanage R, Rath NC. Production and characterization of avian crypt-villus enteroids and the effect of chemicals. BMC Vet Res. 2020;16(1):179.
  7. Alfajaro MM, Kim JY, Barbe L, Cho EH, Park JG, Soliman M, et al. Dual Recognition of Sialic Acid and αGal Epitopes by the VP8* Domains of the Bovine Rotavirus G6P[5] WC3 and of Its Mono-reassortant G4P[5] RotaTeq Vaccine Strains. J Virol. 2019;93(18):e00941-19.
  8. Li L, Fu F, Guo S, Wang H, He X, Xue M, et al. Porcine intestinal enteroids: a new model for studying enteric coronavirus porcine epidemic diarrhea virus infection and the host innate response. J Virol. 2019;93(5):e01682-18.
  9. Chua KB, Voon K, Crameri G, Tan HS, Rosli J, McEachern JA, et al. Identification and characterization of a new orthoreovirus from patients with acute respiratory infections. PLoS One. 2008;3(11):e3803.
  10. Wang L, Fu S, Cao L, Lei W, Cao Y, Song J, et al. Isolation and identification of a natural reassortant mammalian orthoreovirus from least horseshoe bat in China. PLoS One. 2015;10(3):e0118598.
  11. Li Z, Shao Y, Liu C, Liu D, Guo D, Qiu Z, et al. Isolation and pathogenicity of the mammalian orthoreovirus MPC/04 from masked civet cats. Infect Genet Evol. 2015;36:55-61. https://doi.org/10.1016/j.meegid.2015.08.037
  12. Anbalagan S, Spaans T, Hause BM. Genome sequence of the novel reassortant mammalian orthoreovirus strain MRV00304/13, isolated from a calf with diarrhea from the United States. Genome Announc. 2014;2(3):e00451-14.
  13. Kwon HJ, Kim HH, Kim HJ, Park JG, Son KY, Jung J, et al. Detection and molecular characterization of porcine type 3 orthoreoviruses circulating in South Korea. Vet Microbiol. 2012;157(3-4):456-463. https://doi.org/10.1016/j.vetmic.2011.12.032
  14. Thimmasandra Narayanappa A, Sooryanarain H, Deventhiran J, Cao D, Ammayappan Venkatachalam B, Kambiranda D, et al. A novel pathogenic Mammalian orthoreovirus from diarrheic pigs and Swine blood meal in the United States. mBio. 2015;6(3):e00593-e15.
  15. Wang L, Li Y, Walsh T, Shen Z, Li Y, Deb Nath N, et al. Isolation and characterization of novel reassortant mammalian orthoreovirus from pigs in the United States. Emerg Microbes Infect. 2021;10(1):1137-1147. https://doi.org/10.1080/22221751.2021.1933608
  16. Yang DK, An S, Park Y, Yoo JY, Park YR, Park J, et al. Isolation and identification of mammalian orthoreovirus type 3 from a Korean roe deer (Capreolus pygargus). Korean J Vet Res. 2021;61(2):e13.
  17. Beaumont M, Blanc F, Cherbuy C, Egidy G, Giuffra E, Lacroix-Lamande S, et al. Intestinal organoids in farm animals. Vet Res. 2021;52(1):33.
  18. van der Hee B, Loonen LM, Taverne N, Taverne-Thiele JJ, Smidt H, Wells JM. Optimized procedures for generating an enhanced, near physiological 2D culture system from porcine intestinal organoids. Stem Cell Res. 2018;28:165-171. https://doi.org/10.1016/j.scr.2018.02.013
  19. Topfer E, Pasotti A, Telopoulou A, Italiani P, Boraschi D, Ewart MA, et al. Bovine colon organoids: from 3D bioprinting to cryopreserved multi-well screening platforms. Toxicol In Vitro. 2019;61:104606.
  20. Zhang M, Lv L, Cai H, Li Y, Gao F, Yu L, et al. Long-term expansion of porcine intestinal organoids serves as an in vitro model for swine enteric coronavirus infection. Front Microbiol. 2022;13:865336.
  21. Yin L, Chen J, Li L, Guo S, Xue M, Zhang J, et al. Aminopeptidase N expression, not interferon responses, determines the intestinal segmental tropism of porcine deltacoronavirus. J Virol. 2020;94(14):e00480-20.
  22. Resende TP, Medida RL, Vannucci FA, Saqui-Salces M, Gebhart C. Evaluation of swine enteroids as in vitro models for Lawsonia intracellularis infection1,2. J Anim Sci. 2020;98(2):skaa011.
  23. Tong T, Qi Y, Bussiere LD, Wannemuehler M, Miller CL, Wang Q, et al. Transport of artificial virus-like nanocarriers through intestinal monolayers via microfold cells. Nanoscale. 2020;12(30):16339-16347. https://doi.org/10.1039/D0NR03680C
  24. Qin P, Li H, Wang JW, Wang B, Xie RH, Xu H, et al. Genetic and pathogenic characterization of a novel reassortant mammalian orthoreovirus 3 (MRV3) from a diarrheic piglet and seroepidemiological survey of MRV3 in diarrheic pigs from east China. Vet Microbiol. 2017;208:126-136. https://doi.org/10.1016/j.vetmic.2017.07.021
  25. Bagga S, Bouchard MJ. Cell Cycle Regulation During Viral Infection. New York: Springer; 2014.
  26. Bomidi C, Robertson M, Coarfa C, Estes MK, Blutt SE. Single-cell sequencing of rotavirus-infected intestinal epithelium reveals cell-type specific epithelial repair and tuft cell infection. Proc Natl Acad Sci U S A. 2021;118(45):e2112814118.
  27. Jung K, Annamalai T, Lu Z, Saif LJ. Comparative pathogenesis of US porcine epidemic diarrhea virus (PEDV) strain PC21A in conventional 9-day-old nursing piglets vs. 26-day-old weaned pigs. Vet Microbiol. 2015;178(1-2):31-40. https://doi.org/10.1016/j.vetmic.2015.04.022
  28. Xiao Y, Zhou Y, Sun S, Wang H, Wu S, Bao W. Effect of promoter methylation on the expression of porcine MUC2 gene and resistance to PEDV infection. Front Vet Sci. 2021;8:646408.
  29. Rose-John S, Winthrop K, Calabrese L. The role of IL-6 in host defence against infections: immunobiology and clinical implications. Nat Rev Rheumatol. 2017;13(7):399-409. https://doi.org/10.1038/nrrheum.2017.83
  30. Jeffery V, Goldson AJ, Dainty JR, Chieppa M, Sobolewski A. IL-6 signaling regulates small intestinal crypt homeostasis. J Immunol. 2017;199(1):304-311. https://doi.org/10.4049/jimmunol.1600960
  31. Dai J, Pan W, Wang P. ISG15 facilitates cellular antiviral response to dengue and West Nile virus infection in vitro. Virol J. 2011;8(1):468.
  32. He XM, Du X, Zhuo JS, Jing XY, Yang XQ, Liu D. Promoter identification and effect on activation of NF-κB of porcine ISG58. Acta Agric Scand A Anim Sci. 2017;67(1-2):40-45. https://doi.org/10.1080/09064702.2017.1341952