Summary
The development of rotavirus vaccines against acute gastroenteritis for human infants has been accorded a very high priority. Several vaccine candidates all of which are live cultivated strains of animal origin have been tested in humans. However the nature of attenuation of these viruses for humans is unknown. In this study we have attenuated a pig rotavirus by 15 sequential passages in cell culture after which the virus no longer causes diarrhoea in piglets. The pathogenesis of infection of the attenuated rotavirus strain (AT/76 P15) in gnotobiotic piglets was compared with that of the virulent parent strain (AT/76). The pattern of virus replication in the small intestine was judged by histology, disaccharidase assay, immunoperoxidase labelling of gut sections using group A specific rotavirus antibody, and rotavirus antigen assay of gut contents. The parent strain caused variable but extensive infection that resulted in the complete destruction of mature small intestinal enterocytes and villous contraction within 3 days. Membrane bound digestive enzymes were lost, and profound watery diarrhoea and dehydration resulted in causing piglets to become moribund. In contrast attenuated virus appeared to propagate at a much slower pace. Fewer infected epithelial cells were detected at any one time. Destruction of enterocytes was never extensive enough to cause marked mucosal changes in histology. Membrane bound digestive enzymes remained near normal levels and there was little or no diarrhoea. Virus replication ceased after 6 days.
It is concluded that attenuation of the porcine rotavirus strain studied was associated with its decreased ability to propagate in enterocytes after adaption to culture.
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Tzipori S (1985) The relative importance of enteric pathogens affecting neonates of domestic animals. Adv Vet Sci Comp Med 29: 103–206
Wyatt RG, James HD, 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
Hoshino Y, Wyatt RG, Greenberg HB (1984) Serotypic similarity and diversity of rotaviruses of mammalian and avian origin as studied by plaque reduction neutralization. J Infect Dis 149: 694–702
Bridger JC, Pocock DH (1986) Variation in virulence of bovine rotaviruses. J Hyg 96: 257–264
Institute of Medicine (1986) Prospects for immunizing against rotavirus. In: New vaccine development: establishing priorities, vol 2, diseases of importance in developing countries. National Academy Press, Washington, DC, D-13–1–12
Midthun K, Greenberg HB, Hoshino Y, Kapikian AZ, Chanock RM (1985) Reassortant rotaviruses as potential live rotavirus vaccine candidates. J Virol 53: 949–954
Clark HF, Furukawa R, Bell LM, Offit PA, Perrella PA, Plotkin SA (1986) Immune response of infants and children to low-passage bovine rotavirus (strain WC3). Am J Dis Child 140: 350–356
Fernelius AL, Ritchie AE, Classick LG, Norman JO, Mebus CA (1972) Cell culture adapation and propagation of a reovirus-like agent of calf diarrhea from a field outbreak in Nebraska. Arch Ges Virusforsch 37: 114–130
Vesikari T, Isolauri E, D'Hondt E, Delem A, Andre FE, Zissis G (1984) Protection of infants against rotavirus diarrhoea by RIT 4237 attenuated bovine rotavirus strain vaccine. Lancet 1: 977–981
Woode GN, Bew ME, Dennis MJ (1978) Studies on cross-protection induced in calves by rotaviruses from calves, children and fowls. Vet Rec 103: 32–34
Mebus CA, Wyatt RG, Sharpee KS, Sereno MM, Kalica AR, Kapikian AZ, Twiehaus MT (1976) Diarrhoea in gnotobiotic calves caused by reovirus-like agent of human infantile gastroenteritis. Infect Immun 14: 471–479
Tzipori S, Makin T, Smith M (1980) The clinical response of gnotobiotic calves, pigs and lambs to inoculation with human, calf, pig and foal rotavirus isolates. Aust J Exp Biol Med Sci 58: 309–318
Bishop RF, Barnes GL, Cipriani E, Lund JS (1983) Clinical immunity after neonatal rotavirus infection. A prospective longitudinal study in young children. N Engl J Med 309: 72–76
Chrystie IL, Totterdell B, Banatvala JE (1978) Asymptomatic endemic rotavirus infections in the newborn. Lancet 1: 1176–1178
Perez-Schael I, Daoud G, White L, Urbina G, Daoud N, Perez M, Flores J (1984) Rotavirus shedding by newborn children. J Med Virol 14: 127–136
Tzipori S, Williams IH (1978) Diarrhoea in piglets inoculated with rotavirus. Aust Vet J 54: 188–192
Smith M, Tzipori S (1979) Electrophoresis of rotavirus RNA derived from 6 different animal species. Aust J Exp Biol Med Sci 57: 583–585
Albert MJ, Unicomb LE, Tzipori S, Bishop RF (1987) Isolation and serotyping of animal rotaviruses and antigenic comparison with human rotaviruses. Arch Virol 93: 123–130
Makin T, Tzipori S (1980) Inexpensive technique for the production and maintenance of gnotobiotic piglets, calves and lambs. Aust Vet J 56: 353–359
Tzipori S, Robins-Browne RM, Gonis G, Hayes J, Whithers M, McCartney E (1984) EnteropathogenicEscherichia coli enteritis: evaluation of the gnotobiotic piglet as a model of human infection. Gut 26: 570–578
Coulson BS, Unicomb LE, Pitson GA, Bishop RF (1987) Simple and specific enzyme immunoassay using monoclonal antibodies for serotyping human rotaviruses. J Clin Microbiol 25: 509–515
Dahlquist A (1964) Method for assay of intestinal disaccharidases. Anal Biochem 7: 18–25
Anderson EL, Belshe RB, Bartram J, Crookshanks-Newman F, Chanock RM, Kapikian AZ (1986) Evaluation of rhesus rotavirus vaccine (MMU 18006) in infants and young children. J Infect Dis 153: 823–831
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Tzipori, S., Unicomb, L., Bishop, R. et al. Studies on attenuation of rotavirus. Archives of Virology 109, 197–205 (1989). https://doi.org/10.1007/BF01311081
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DOI: https://doi.org/10.1007/BF01311081