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HT-29 cells: a new substrate for rotavirus growth

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Summary

Susceptibilities of a continuous rhesus monkey kidney cell line (MA-104) and that of a human colon carcinoma cell line (HT-29) to infection by different human and animal rotavirus strains were compared.

HT-29 cells appeared to be more sensitive to human rotavirus infection than MA-104 cells, whereas the latter cell line was more susceptible to animal rotavirus replication. The greater sensitivity to human rotavirus infection of HT-29 cells was confirmed by the successful, direct isolation of these viruses from faecal specimens. Human rotavirus infection of HT-29 cells was also followed by transmission electron microscopy. In ultra-thin sections, unenveloped particles of rotaviruses, representing infectious mature virions, were observed in large number. Moreover, many “double-shelled” particles were detected in negative-stained supernatants from infected cultures. Scanning electron microscopy of uninfected HT-29 cells showed that in the presence of Ca+ +, required for rotavirus growth, they are able to express some of the features of mature intestinal cells.

In view of these results, HT-29 cells appear to be a useful in vitro model for the study of rotavirus infection.

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References

  1. Albert MJ, Bishop RF (1984) Cultivation of human rotaviruses in cell culture. J Med Virol 13: 377–383

    Google Scholar 

  2. Albert MJ, Unicomb LE, Barnes GL, Bishop RF (1987) Cultivation and characterization of rotavirus strains infecting newborn babies in Melbourne, Australia, from 1975 to 1979. J Clin Microbiol 25: 1635–1640

    Google Scholar 

  3. Bastardo JW, McKimm-Breschkin JL, Sonza S, Mercer LD, Holmes IH (1981) Preparation and characterization of antisera to electrophoretically purified SA-11 virus polypeptides. Infect Immun 34: 641–647

    Google Scholar 

  4. Birch CJ, Rodger SM, Marshall JA, Gust ID (1983) Replication of human rotavirus in cell culture. J Med Virol 11: 241–250

    Google Scholar 

  5. Brandt CD, Kim HW, Rodriguez WJ, Arrobio JO, Jeffries BC, Stallings EP, Lewis C, Miles AJ, Chanock RM, Kapikian AZ, Parrott RH (1983) Pediatric viral gastroenteritis during eight years of study. J Clin Microbiol 18: 71–78

    Google Scholar 

  6. Bridger JC, Woode GN (1976) Characterization of two particle types of calf rotavirus. J Gen Virol 31: 245–250

    Google Scholar 

  7. Chasey D (1977) Different particle types in tissue culture and intestinal epithelium infected with rotavirus. J Gen Virol 37: 443–451

    Google Scholar 

  8. Davidson GP, Gall DG, Petric M, Butler DG, Hamilton JR (1977) Human rotavirus enteritis induced in conventional piglets: intestinal structure and transport. J Clin Invest 60: 1402–1409

    Google Scholar 

  9. Donelli G, Ruggeri FM, Tinari A, Marziano ML, Menichella D, Caione D, Concato C, Rocchi G, Vella S (1988) A three-year diagnostic and epidemiological study on viral infantile diarrhoea in Rome. Epidemiol Infect 100: 311–320

    Google Scholar 

  10. Estes MK (1990) Rotaviruses and their replication. In: Fields BN (ed) Virology, vol 2, 2nd edn. Raven, New York, pp 1329–1352

    Google Scholar 

  11. Hasegawa A, Matsuno S, Inouye S, Kono R, Tsurukubo Y, Mukoyama A, Saito Y (1982) Isolation of human rotaviruses in primary cultures of monkey kidney cells. J Clin Microbiol 16: 387–390

    Google Scholar 

  12. Huet C, Sahuquillo-Merino C, Coudrier E, Louvard D (1987) Absorptive and mucus-secreting subclones isolated from a multipotent intestinal cell line (HT-29) provide new models for cell polarity and terminal differentiation. J Cell Biol 105: 345–357

    Google Scholar 

  13. Kapikian AZ, Wyatt RG, Greenberg HB, Kalica AR, Kim HW, Brandt CD, Rodriguez WJ, Parrott RH, Chanock RM (1980) Approaches to immunization of infants and young children against gastroenteritis due to rotaviruses. Rev Infect Dis 2: 459–469

    Google Scholar 

  14. Kapikian AZ, Greenberg HB, Wyatt RG, Kalica AR, Kim HW, Brandt CD, Rodriguez WJ, Parrott RH, Chanock RM (1982) Viral gastroenteritis. In: Evans AS (ed) Viral infections of humans, 2nd edn. Plenum, New York, pp 283–236

    Google Scholar 

  15. Kapikian AZ, Chanock RM (1990) Rotaviruses. In: Fields BN (ed) Virology, vol 2, 2nd edn. Raven, New York, pp 1353–1404

    Google Scholar 

  16. Kerzner B, Kelly MH, Gall DG, Butler DG, Hamilton JR (1977) Transmissible gastroenteritis: sodium transport in the intestinal epithelium during the course of viral enteritis. Gastroenterology 72: 457–461

    Google Scholar 

  17. Konno T, Suzuki H, Imai A, Kutsuzawa T, Ishida N, Katsushima N, Sakamoto M, Kitaoka S, Tsuboi R, Adachi M (1978) A long-term survey of rotavirus infection in Japanese children with acute gastroenteritis. J Infect Dis 138: 569–576

    Google Scholar 

  18. Kutsuzawa T, Konno T, Suzuki H, Kapikian AZ, Ebina T, Ishida N (1982) Isolation of human rotavirus subgroups 1 and 2 in cell cultures. J Clin Microbiol 16: 727–730

    Google Scholar 

  19. Ludert JE, Michelangeli F, Gil F, Liprandi F, Esparza J (1987) Penetration and uncoating of rotaviruses in cultured cells. Intervirology 27: 95–101

    Google Scholar 

  20. McNulty MS (1976) The morphogenesis of a cytopathic bovine rotavirus in Madin-Darby bovine kidney cells. J Gen Virol 33: 503–508

    Google Scholar 

  21. Naguib T, Wyatt RG, Mohieldin MS, Zaki AM, Imam IZ, DuPont HL (1984) Cultivation and subgroup determination of human rotaviruses from Egyptian infants and young children. J Clin Microbiol 19: 210–212

    Google Scholar 

  22. Patel JR, Daniel J, Mathan VI (1985) A comparison of the susceptibility of three human gut tumour-derived differentiated epithelial cell lines, primary monkey kidney cells and human rhabdomyosarcoma cell line to 66 prototype strains of human enterovirus. J Virol Methods 12: 209–216

    Google Scholar 

  23. Petrie BL, Graham DY, Estes MK (1981) Identification of rotavirus particle types. Intervirology 16: 20–28

    Google Scholar 

  24. Petrie BL, Graham DY, Haussen H, Estes MK (1982) Localization of rotavirus antigens in infected cells by ultrastructural immunocytochemistry. J Gen Virol 63: 457–467

    Google Scholar 

  25. Petrie BL, Greenberg HB, Graham DY, Estes MK (1984) Ultrastructural localization of rotavirus antigens using colloidal gold. Virus Res 1: 133–152

    Google Scholar 

  26. Pinto M, Appay MD, Simon-Assmann P, Chevalier G, Dracopoli N, Fogh J, Zweibaum A (1982) Enterocytic differentiation of cultured human colon cancer cells by replacement of glucose by galactose in the medium. Biol Cell 44: 193–196

    Google Scholar 

  27. Quaroni A, May RJ (1980) Establishment and characterization of intestinal epithelial cell cultures. In: Harris GC, Trump BF, Stoner GD (eds) Methods in cell biology, vol 21 B. Academic Press, New York, pp 403–427

    Google Scholar 

  28. Quaroni A (1985) Development of fetal rat intestine in organ and monolayer culture. J Cell Biol 100: 1601–1610

    Google Scholar 

  29. Riepenhoff-Talty M, Saif LJ, Barrett HJ, Suzuki H, Ogra PL (1983) Potential spectrum of etiological agents of viral gastroenteritis in hospitalized infants. J Clin Microbiol 17: 352–356

    Google Scholar 

  30. Robine S, Huet C, Moll R, Sahuquillo-Merino C, Coudrier E, Zweibaum A, Louvard D (1985) Can villin be used to identify malignant and undifferentiated normal digestive epithelial cells? Proc Natl Acad Sci USA 82: 8488–8492

    Google Scholar 

  31. Rousset M (1986) The human colon carcinoma cell lines HT-29 and Caco-2: two in vitro models for the study of intestinal differentiation. Biochimie 68: 1035–1040

    Google Scholar 

  32. Sato K, Inaba Y, Shinozaki T, Fujii R, Matumotu M (1981) Isolation of human rotavirus in cell cultures. Arch Virol 69: 155–160

    Google Scholar 

  33. Shahrabadi MS, Lee PWK (1986) Bovine rotavirus maturation is a calcium-dependent process. Virology 152: 298–307

    Google Scholar 

  34. Shahrabadi MS, Babink LA, Lee PWK (1986) Further analysis of the role of calcium in rotavirus morphogenesis. Virology 158: 103–111

    Google Scholar 

  35. Shepherd RW, Gall DG, Butler DG, Hamilton JR (1979) Determinants of diarrhea in viral enteritis: the role of ion transport and epithelial changes in ileum in trasmissible gastroenteritis in piglets. Gastroenterology 76: 20–24

    Google Scholar 

  36. Shirley JA, Beards GM, Thouless ME, Flewett TH (1981) The influence of divalent cations on the stability of human rotavirus. Arch Virol 67: 1–9

    Google Scholar 

  37. Snodgrass DR, Angus KW, Gray EW (1977) Rotavirus infection in lambs: pathogenesis and pathology. Arch Virol 55: 263–274

    Google Scholar 

  38. Tinari A, Ruggeri FM, Divizia M, Panà A, Donelli G (1989) Morphological changes in HAV-infected Frp/3 cells and immunolocalization of HAAg. Arch Virol 104: 209–224

    Google Scholar 

  39. Tufvesson B, Johnsson T (1976) Occurrence of reo-like viruses in young children with acute gastroenteritis. Acta Pathol Microbiol Scand 84: 22–28

    Google Scholar 

  40. Urasawa T, Urasawa S, Taniguchi K (1981) Sequential passages of human rotavirus in MA-104 cells. Microbiol Immunol 25: 1025–1035

    Google Scholar 

  41. Ward RL, Knowlton DR, Pierce MJ (1984) Efficiency of human rotavirus propagation in cell culture. J Clin Microbiol 19: 748–753

    Google Scholar 

  42. Wyatt RG, James HD jr, Pittman AL, Hoshino Y, Greenberg HB, Kalica AR, Flores J, Kapikian AZ (1983) Direct isolation in cell culture of human rotaviruses and their characterization into four serotypes. J Clin Microbiol 18: 310–317

    Google Scholar 

  43. Zweibaum A, Pinto M, Chevalier G, Dussaulx E, Triadou N, Sacroix B, Haffen K, Brun JL, Rousset M (1985) Enterocytic differentiation of a subpopulation of the human colon tumour cell line HT-29 selected for growth in sugar-free medium and its inhibition by glucose. J Cell Physiol 122: 21–29

    Google Scholar 

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  1. Laboratorio di Ultrastrutture, Istituto Superiore di Sanità, Roma, Italy

    F. Superti, A. Tinari, L. Baldassarri & G. Donelli

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  1. F. Superti
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Superti, F., Tinari, A., Baldassarri, L. et al. HT-29 cells: a new substrate for rotavirus growth. Archives of Virology 116, 159–173 (1991). https://doi.org/10.1007/BF01319239

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