Application of Immunohistochemistry to Study of Avian Leukosis Virus Free

Abstract

Summary

A modification of the peroxidase-labelled antibody technique was applied to study the distribution of virus antigens in chicken cells infected with two serotypes (A and B) of avian leukosis virus (ALV). Type-specific chicken antisera reacted only with cells infected with virus of the homologous envelope serotype. When unfixed cells were exposed to type-specific antivirus serum, only antigens located at the cell surface were stained, while cells exposed to type-specific antibodies after fixation revealed both surface and intracytoplasmic virus antigen. Cytoplasmic antigen was usually concentrated in discrete granules which often had a vesicular structure.

Hamster antibodies against ALV group-specific () antigen reacted with fixed infected cells regardless of envelope serotype, and the distribution of antigen was as shown with chicken type-specific antibodies. Intranuclear antigen may have been present in a few cells. Unfixed infected cells did not react with hamster antibody and confirmed the location of ALV antigen within the virus particle.

Some chicken antisera with neutralizing antibodies against a single envelope serotype of ALV contained both type-specific and antibodies. These antisera reacted with fixed cells infected with virus of either the homologous or heterologous serotype and stained both surface and cytoplasmic antigens. With unfixed infected cells, these antisera combined only with the surface of cells infected with virus of the same serotype as the neutralizing antibodies in the serum. Thus, the reaction in heterologously infected fixed cells was with internal antigen. This confirms independently that chickens are not naturally tolerant to their homologous ‘C type’ antigens.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-15-2-149
1972-05-01
2024-03-28
Loading full text...

Full text loading...

/deliver/fulltext/jgv/15/2/JV0150020149.html?itemId=/content/journal/jgv/10.1099/0022-1317-15-2-149&mimeType=html&fmt=ahah

References

  1. Abelson H. T., Smith G. H., Hoffman H. A., Rowe W. P. 1969; Use of enzyme-labeled antibody for electron microscope localization of lymphocytic choriomeningitis virus antigens in infected cell cultures. Journal of the National Cancer Institute 42:497
    [Google Scholar]
  2. Armstrong D. 1969; Group-specific components of avian tumor viruses detected with chicken and hamster sera. Journal of Virology 3:133
    [Google Scholar]
  3. Avrameas S. 1969; Coupling of enzymes to proteins with glutaraldehyde. Use of the conjugates for the detection of antigens and antibodies. Immunochemistry 6:43
    [Google Scholar]
  4. Benedict A. A. 1967; Purification of chicken immunoglobulins. In Methods in Immunology and Immunochemistry vol 1 p 233 Ed. by Williams C. A., Chase M. W. New York: Academic Press;
    [Google Scholar]
  5. Dougherty R. M., Distefano H. S. 1966; Lack of relationship between infection with avian leukosis virus and presence of COFAL antigen in chick embryos. Virology 29:586
    [Google Scholar]
  6. Dougherty R. M., Distefano H. S., Roth F. K. 1967; Virus particles and viral antigens in chicken tissues free of infectious avian leukosis virus. Proceedings of the National Academy of Sciences of the United States of America 58:808
    [Google Scholar]
  7. Dougherty R. M., Rasmussen R. 1964; Properties of a strain of Rous sarcoma virus that infects mammals. National Cancer Institute Monograph 17:337
    [Google Scholar]
  8. Fink M. A., Malmgren R. A. 1963; Fluorescent antibody studies of the viral antigen in a murine leukemia (Rauscher). Journal of the National Cancer Institute 31:1111
    [Google Scholar]
  9. Goldstein G., Slizys S., Chase M. W. 1961; Studies on fluorescent antibody Staining. I. Non-specific fluorescence with fluorescein-coupled sheep anti-rabbit globulins. Journal of Experimental Medicine 114:89
    [Google Scholar]
  10. Goodman M., Wolfe H. R., Norton S. 1951; Precipitin production in chickens. IV. The effect of varying concentrations of NaCl on precipitate formation. Journal of Immunology 66:225
    [Google Scholar]
  11. Graham R. C. Jun, Karnovsky M. J. 1966; The early stages of absorption of injected horseradish peroxidase in the proximal tubules of mouse kidney: Ultrastructural cytochemistry by a new technique. Journal of Histochemistry and Cytochemistry 14:291
    [Google Scholar]
  12. Hanafusa H., Miyamoto T., Hanafusa T. 1970; A cell-associated factor essential for formation of an infectious form of Rous sarcoma virus. Proceedings of the National Academy of Sciences of the United States of America 66:314
    [Google Scholar]
  13. Hanafusa T., Hanafusa H., Miyamoto T. 1970; Recovery of a new virus from apparently normal chick cells by infection with avian tumor viruses. Proceedings of the National Academy of Sciences of the United States of America 67:1797
    [Google Scholar]
  14. Huebner R. J., Sarma P. S., Kelloff G. J., Gilden R. V., Meier H., Peters R. L. 1970; Immunological tolerance to RNA tumor virus genome expressions: Significance of tolerance and prenatal expressions in embryogenesis and tumorigenesis. Annals of the New York Academy of Sciences 181:246
    [Google Scholar]
  15. Huebner R. J., Todaro G. J. 1969; Oncogenes of RNA tumor viruses as determinants of cancer. Proceedings of the National Academy of Sciences of the United States of America 64:1087
    [Google Scholar]
  16. Ishizaki R., Vogt P. K. 1966; Immunological relationships among envelope antigens and avian tumor viruses. Virology 30:375
    [Google Scholar]
  17. Kelloff G., Vogt P. K. 1966; Localization of avian tumor virus group specific antigen in cell and virus. Virology 29:377
    [Google Scholar]
  18. Klein E., Klein G. 1964; Antigenic properties of lymphomas induced by the Moloney agent. Journal of the National Cancer Institute 32:547
    [Google Scholar]
  19. Nakane P. K., Pierce G. B. 1967; Enzyme-labeled antibodies for light and electron microscopic localization of tissue antigens. Journal of Cell Biology 33:307
    [Google Scholar]
  20. Payne F. E., Solomon J. J., Purchase H. G. 1966; Immunofluorescent studies of group-specific antigen of the avian sarcoma-leukosis viruses. Proceedings of the National Academy of Sciences of the United States of America 55:341
    [Google Scholar]
  21. Payne L. N., Chubb R. C. 1968; Studies on the nature and genetic control of an antigen in normal chick embryos which reacts in the COFAL test. Journal of General Virology 3:379
    [Google Scholar]
  22. Peters T. Jun, Ashley A. A. 1967; An artifact in radioautography due to binding of free amino acids to tissues by fixatives. Journal of Cell Biology 33:53
    [Google Scholar]
  23. Rabotti G. F., Blackham E. 1970; Immunological determinants of avian sarcoma viruses: Presence of group-specific antibodies in fowl sera demonstrated by complement-fixation inhibition test. Journal of the National Cancer Institute 44:985
    [Google Scholar]
  24. Rice C. E. 1948; Some factors influencing selection of complement-fixation method; parallel use of direct and indirect techniques. Journal of Immunology 60:11
    [Google Scholar]
  25. Roth F. K., Meyers P., Dougherty R. M. 1971; The presence of avian leukosis group-specific antibodies in chicken sera. Virology 45:265
    [Google Scholar]
  26. Roth F. K., Dougherty R. M. 1969; Multiple antigenic components of the group-specific antigen of the avian leukosis-sarcoma viruses. Virology 38:278
    [Google Scholar]
  27. Rubin H., Vogt P. K. 1962; An avian leukosis virus associated with stocks of Rous sarcoma virus. Virology 17:184
    [Google Scholar]
  28. Singer S. J., Schick A. F. 1961; The properties of specific stains for electron microscopy prepared by the conjugation of antibody molecules with ferritin. Journal of Biochemical and Biophysical Cytology 9:519
    [Google Scholar]
  29. Sternberger L., Hardy P. H., Cuculis J. J., Meyer H. G. 1970; The unlabeled antibody enzyme method of immunohistochemistry. Preparation and properties of soluble antigen-antibody complex (horseradish peroxidase-anti-horseradish peroxidase) and its use in identification of spirochetes. Journal of Histochemistry and Cytochemistry 18:315
    [Google Scholar]
  30. Vogt P. K., Friis R. R. 1971; An avian leukosis virus related to RSV (O): Properties and evidence for helper activity. Virology 43:223
    [Google Scholar]
  31. Vogt P. K., Ishizaki R. 1965; Reciprocal patterns of genetic resistance to avian tumor viruses in two lines of chickens. Virology 26:664
    [Google Scholar]
  32. Vogt P. K., Lukyx N. 1963; Observations on the surface of cells infected with Rous sarcoma virus. Virology 20:75
    [Google Scholar]
  33. Vogt P. K., Rubin H. 1961; Localization of infectious virus and viral antigen in chick fibroblasts during successive stages of infection with Rous sarcoma virus. Virology 13:528
    [Google Scholar]
  34. Weiss R. A. 1969; The host range of Bryan strain Rous sarcoma virus synthesized in the absence of helper virus. Journal of General Virology 5:511
    [Google Scholar]
  35. Worthington Biochemical Corporation 1966; Enzyme catalogue. Freehold, New Jersey.:
    [Google Scholar]
  36. Yohn D. S., Weber J., Mccammon J. R. 1971; Avian leukosis group-specific (ALG-S) antibodies in COFAL-negative sera. Proceedings of the American Association for Cancer Research 12:22
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-15-2-149
Loading
/content/journal/jgv/10.1099/0022-1317-15-2-149
Loading

Data & Media loading...

Most cited Most Cited RSS feed