1887

Abstract

Summary

Pulse labelling of cells with [S]methionine at different times after infection followed by SDS-PAGE was used to resolve and to identify polypeptides designated as specific to transmissible gastroenteritis virus (TGEV)-infected swine testicular (ST) cell cultures. The major TGEV structural proteins, with apparent molecular weights of 200000 (200K), 47K and 30K were detected in radiolabelled cell extracts by 6 h post-infection. Additionally, a 17K major polypeptide was present in infected cells but not in mock-infected control cultures. Labelling with [H]glucosamine revealed only the 200K and 30K proteins to be glycosylated. TGEV-primed porcine lymphocytes, secondarily stimulated with sucrose gradient-purified virus, produced antibody only to the two glycoproteins (gp) indicating that the 17K polypeptide is not a surface feature of the virion. Two pigs were infected oronasally with the virulent Miller strain of TGEV and their sera were analysed by immunoprecipitation. At 25 days post-infection convalescent sera responded strongly to gp30 and gp200 and there was a weak initial response to the 17K polypeptide. Serum immunoglobulins at 60 days post-infection reacted strongly to the 17K protein while the antibody response to gp30 was significantly reduced and that to gp200 was slightly reduced.

Keyword(s): 17K antigen , immune response and TGEV
Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-67-7-1419
1986-07-01
2022-08-15
Loading full text...

Full text loading...

/deliver/fulltext/jgv/67/7/JV0670071419.html?itemId=/content/journal/jgv/10.1099/0022-1317-67-7-1419&mimeType=html&fmt=ahah

References

  1. Bayliss G. J., Deby G., Wolf H. 1983; An immunoprecipitation blocking assay for the analysis of EBV induced antigens. Journal of Virological Methods 7:229–239
    [Google Scholar]
  2. Bohl E. H. 1981; Transmissible gastroenteritis. In Diseases of Swine 5th edn pp 195–208 Edited by Leman D. A., Glock R. D., Mengeling W. L., Penny R. H. C., Scholl E., Straw B. Ames: Iowa State University Press;
    [Google Scholar]
  3. Boursnell M. E. G., Binns M. M., Brown T. D. K. 1985; Sequencing of coronavirus IBV genomic RNA: three open reading frames in the 5′ ‘unique’ region of mRNA D. Journal of General Virology 66:2253–2258
    [Google Scholar]
  4. Brian D. A., Hogue B., Lapps W., Potts B., Kapke P. 1984; Comparative structure of coronaviruses. In Proceedings, Fourth International Symposium on Neonatal Diarrhea pp 100–116 Saskatoon: Veterinary Infectious Disease Organization;
    [Google Scholar]
  5. Cavanagh D. 1981; Structural polypeptides of coronavirus IBV. Journal of General Virology 53:93–103
    [Google Scholar]
  6. Chamberlain J. P. 1979; Fluorographic detection of radioactivity in polyacrylamide gels with the water-soluble fluor, sodium salicylate. Analytical Biochemistry 98:132–135
    [Google Scholar]
  7. Furuuchi S., Shimuzu Y., Kumagai T. 1978/9; Multiplication of low and high cell culture passaged strains of transmissible gastroenteritis virus in organs of newborn piglets. Veterinary Microbiology 3:169–178
    [Google Scholar]
  8. Garwes D. J., Pocock D. H. 1975; The polypeptide structure of transmissible gastroenteritis virus. Journal of General Virology 29:25–34
    [Google Scholar]
  9. Horzinek M. C., Lutz H., Pedersen N. C. 1982; Antigenic relationships among homologous structural polypeptides of porcine, feline, and canine coronaviruses. Infection and Immunity 37:1148–1155
    [Google Scholar]
  10. Hu S., Bruszewski J., Boone T., Souza L. 1984; Cloning and expression of the surface glycoprotein gp 195 of porcine transmissible gastroenteritis virus. In Modern Approaches to Vaccines pp 219–223 Edited by Chanock R. M., Lerner R. A. New York: Cold Spring Harbor Laboratory;
    [Google Scholar]
  11. Laemmli U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, London 227:680–685
    [Google Scholar]
  12. Laude H., Charley B., Gelfi J. 1984; Replication of transmissible gastroenteritis coronavirus (TGEV) in swine alveolar macrophages. Journal of General Virology 65:327–332
    [Google Scholar]
  13. Lomniczi B., Morser J. 1981; Polypeptides of infectious bronchitis virus. I. Polypeptides of the virion. Journal of General Virology 55:155–164
    [Google Scholar]
  14. Mcclurkin A. W., Norman J. O. 1966; Studies on transmissible gastroenteritis of swine. II. Selected characteristics of a cytopathogenic virus common to five isolates from transmissible gastroenteritis. Canadian Journal of Comparative Medicine and Veterinary Science 30:190–198
    [Google Scholar]
  15. Siddell S., Wege H., Barthel A., Ter Meulen V. 1981; Coronavirus JHM: intracellular protein synthesis. Journal of General Virology 53:145–155
    [Google Scholar]
  16. Siddell S. G., Anderson R., Cavanagh D., Fujiwara K., Klenk H. D., Macnaughton M. R., Pensaert M., Stohlman S. A., Sturman L., Van Der Zeijst B. A. M. 1983; Coronaviridae. Intervirology 20:181–189
    [Google Scholar]
  17. Skinner M. A., Siddell S. G. 1985; Coding sequence of coronavirus MHV-JHM mRNA 4. Journal of General Virology 66:593–596
    [Google Scholar]
  18. Stern D. F., Burgess L., Sefton B. M. 1982; Structural analysis of virion proteins of the avian coronavirus infectious bronchitis virus. Journal of Virology 42:208–219
    [Google Scholar]
  19. Underdahl N. R., Mebus C. A., Torres-Medina A. 1975; Recovery of transmissible gastroenteritis virus from chronically infected experimental pigs. American Journal of Veterinary Research 36:1473–1476
    [Google Scholar]
  20. Wesley I. V., Koster M. J., Knudsen R. C. 1984; A hemolytic plaque assay for foot-and-mouth disease virus. Annals of the New York Academy of Sciences 435:361–363
    [Google Scholar]
  21. Wesley R. D., Wesley I. V., Paul P. S., Hall M. R., Woods R. D. 1986; Analysis of the humoral immune response in vitro to transmissible gastroenteritis virus. Annals of the New York Academy of Sciences 463:412414
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-67-7-1419
Loading
/content/journal/jgv/10.1099/0022-1317-67-7-1419
Loading

Data & Media loading...

Most cited this month Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error