1887

Abstract

A nucleoprotein (NP) preparation purified from the chorioallantoic membrane of chicken eggs infected with fowl plague virus (A/FPV/Rostock/34, H7N1) yielded, in addition to the commonly known 56K protein, a 42K component that could not be detected in virus particles. After testing with a series of NP-specific monoclonal antibodies it was found that some reacted with both proteins and others were bound only by the 56K protein. Among both types of NP-specific monoclonal antibodies only a limited number were bound to infected murine cells. Some antibodies bound to cells infected with a given subtype failed to react with the surface of cells infected with a different subtype. Binding was demonstrated by cellular ELISA, radioimmunoassay and immunofluorescence. The results indicate that only restricted antigenic domains of the native NP and perhaps NP fragments are exposed at the surface of infected murine cells. Additionally, the purified NP preparation was used to immunize mice in order to determine the protective capacity of cell- associated NP. In parallel, and as a relevant control, mice were immunized with a vaccinia virus recombinant containing the gene for NP prior to challenge with infectious virus. High levels of monospecific antibodies and a cytotoxic T cell activity was found in mice immunized with purified NP or infected with the vaccinia recombinant after secondary restimulation . After treatment with specific antibodies the cytotoxic cells were shown to be classical CD8 cytotoxic T lymphocytes. Despite the elicitation of a humoral and a cellular immune response by the forms of NP employed mice were not protected from influenza virus infection.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-71-5-1169
1990-05-01
2022-11-27
Loading full text...

Full text loading...

/deliver/fulltext/jgv/71/5/JV0710051169.html?itemId=/content/journal/jgv/10.1099/0022-1317-71-5-1169&mimeType=html&fmt=ahah

References

  1. Andrew M. E., Coupar B. E. H. 1988; Efficacy of influenza haemagglutinin and nucleoprotein as protective antigens against influenza virus infection in mice. Scandinavian Journal of Immunology 28:81–85
    [Google Scholar]
  2. Axen R., Ernback S. 1971; Chemical fixation of enzymes to cyanogen halide activated polysaccharide carriers. European Journal of Biochemistry 18:351–360
    [Google Scholar]
  3. Becht H., Malole B. 1975; Comparative evaluation of different fixation procedures and different coupling reagents for the demonstration of influenza virus-specific antibodies by the indirect haemagglutination test. Medical Microbiology and Immunology 162:43–53
    [Google Scholar]
  4. Braciale T. J. 1977; Immunologic recognition of influenza virus- infected cells. I. Generation of a virus-strain specific and a crossreactive subpopulation of cytotoxic T cells in the response to type A influenza viruses of different subtypes. Cellular Immunology 33:423–436
    [Google Scholar]
  5. Cobbold S. P., Jayasuriya A., Nash A., Prospero T. D., Waldmann H. 1984; Therapy with monoclonal antibodies by elimination of T-cell subsets in vivo . Nature; London: 314548–551
    [Google Scholar]
  6. Davenport F. M., Rott R., Schäfer W. 1960; Physical and biological properties of influenza virus components obtained after ether treatment. Journal of Experimental Medicine 112:765–782
    [Google Scholar]
  7. Effros R. B., Doherty P. C., Gerhard W., Bennink J. 1977; Generation of both cross-reactive and virus-specific T-cell populations after immunization with serologically distinct influenza A viruses. Journal of Experimental Medicine 145:557–568
    [Google Scholar]
  8. Fazekas De St Groth U., Scheidegger D. 1980; Production of monoclonal antibodies: strategies and tactics. Journal of Immunological Methods 35:1–20
    [Google Scholar]
  9. Fleischer B., Becht H., Rott R. 1985; Recognition of viral antigens by human influenza A virus-specific T-lymphocyte clones. Journal of Immunology 135:2800–2804
    [Google Scholar]
  10. Hany M., Boehen S., Schulz M., Hengartner H., Mackett M., Bishop D. H. L., Overton H., Zinkernagel R. 1989; Antiviral protection and prevention of lymphocytic choriomeningitis or of local footpad swelling reaction in mice by immunization with vaccinia recombinant virus expressing LCMV-WE nucleoprotein or glycoprotein. European Journal of Immunology 19:417–429
    [Google Scholar]
  11. Hioe C. E., Hinshaw V. S. 1989; Induction and activity of class-II restricted, Lyt-2+ cytolytic T lymphocytes specific for the influenza H5 haemagglutinin. Journal of Immunology 142:2482–2488
    [Google Scholar]
  12. Jensenius J. C., Andersen I., Hau J., Crone M., Koch C. 1981; Eggs: conveniently packaged antibodies. Methods for purification of yolk IgG. Journal of Immunological Methods 46:63–68
    [Google Scholar]
  13. Kearney J. F., Radbruch A., Liesegang B., Rajewsky K. 1979; A new mouse myeloma cell line that lost immunoglobulin expression but permits the construction of antibody secreting hybrid cell lines. Journal of Immunology 123:1548–1550
    [Google Scholar]
  14. Köhler G., Milstein C. 1975; Continuous cultures of cells secreting antibody of predefined specificity. Nature; London: 256495–497
    [Google Scholar]
  15. Lin Y. L., Askonas B. A. 1981; Biological properties of an influenza A virus-specific killer T cell clone. Inhibition of virus replication in vivo and induction of delayed-type hypersensitivity reactions. Journal of Experimental Medicine 154:225–234
    [Google Scholar]
  16. Luckacher A. E., Braciale V. L., Braciale T. J. 1984; In vivo effector function of influenza virus-specific cytotoxic T lymphocyte clones is highly specific. Journal of Experimental Medicine 160:814826
    [Google Scholar]
  17. McMichael A. J., Michie C. A., Gotch F. M., Smith G. L. 1986; Recognition of influenza A virus nucleoprotein by human cytotoxic T lymphocytes. Journal of General Virology 67:719–726
    [Google Scholar]
  18. Morrison L. A., Braciale V. L., Braciale T. J. 1985; Expression of H-2 I region-restricted cytolytic activity by an Lyt-2+ influenza virus-specific T cell clone. Journal of Immunology 135:3691–3698
    [Google Scholar]
  19. Panicali D., Paoletti E. 1982; Construction of poxviruses as cloning vectors: insertion of the thymidine kinase gene from herpes simplex virus into the DNA of infectious vaccinia virus. Proceedings of the National Academy of Sciences U.S.A.: 794927–4931
    [Google Scholar]
  20. Panicali D., Davis S. W., Weinberg R. L., Paoletti E. 1983; Construction of live vaccines by using genetically engineered poxviruses: biological activity of recombinant vaccinia virus expressing influenza virus haemagglutinin. Proceedings of the National Academy of Sciences U.S.A.: 805364–5368
    [Google Scholar]
  21. Porath J., Sundberg L. 1972; High capacity chemosorbents for protein immobilization. Nature New Biology 238:261–262
    [Google Scholar]
  22. Schild G. C., Wood J. M., Newman R. W. 1975; A single-radial- immunodiffusion technique for the assay of influenza haemagglutinin antigen. Bulletin of the World Health Organization 52:223–230
    [Google Scholar]
  23. Stitz L., Huang R. T. C., Hengartner H., Rott R., Zinkernagel R. 1985; Cytotoxic T cell lysis of target cells fused with liposomes containing influenza virus haemagglutinin and neuraminidase. Journal of General Virology 66:1333–1339
    [Google Scholar]
  24. Townsend A. R. M., Skehel J. J. 1982; Influenza A specific cytotoxic T cell clones that do not recognise viral glycoproteins. Nature; London: 300655–657
    [Google Scholar]
  25. Townsend A. R. M., Skehel J. J. 1984; The influenza A virus nucleoprotein gene controls the induction of both subtype specific and crossreactive cytotoxic T cells. Journal of Experimental Medicine 160:552–563
    [Google Scholar]
  26. Townsend A. R. M., McMichael A. J., Carter N. P., Huddles-Tone J. A., Brownlee G. G. 1984; Cytotoxic T cell recognition of the influenza nucleoprotein and hemagglutinin expressed in transfected cells. Cell 39:13–25
    [Google Scholar]
  27. Townsend A. R. M., Gotch F. M., Davey J. 1985; Cytotoxic T cells recognize fragments of the influenza nucleoprotein. Cell 42:457–467
    [Google Scholar]
  28. Townsend A. R. M., Rothbard J., Gotch F. M., Bahadur G., Wraith D., McMichael A. J. 1986; The epitopes of influenza nucleoprotein recognized by cytotoxic T lymphocytes can be defined with short synthetic peptides. Cell 44:959–968
    [Google Scholar]
  29. Van Wyke K., Hackett C. J., Askonas B. A., Webster R. G. 1980; Monoclonal antibodies to influenza matrix protein: detection of low levels of matrix protein on abortively infected cells. Journal of General Virology 47:497–501
    [Google Scholar]
  30. Virelizier J. L., Allison A. C., Oxford J. S., Schild G. C. 1977; Early presence of ribonucleoprotein antigen on surface of influenza virus-infected cells. Nature; London: 26652–54
    [Google Scholar]
  31. Wraith D. C., Askonas B. A. 1985; Induction of influenza A virus cross-reactive cytotoxic T cells by a nucleoprotein/haemagglutinin preparation. Journal of General Virology 66:1327–1331
    [Google Scholar]
  32. Wraith D. C., Vessey A. E., Askonas B. A. 1987; Purified influenza virus nucleoprotein protects mice from lethal infection. Journal of General Virology 68:433–440
    [Google Scholar]
  33. Zhirnov O. P., Bukrinskaya A. G. 1981; Two forms of influenza nucleoprotein in infected cells and virions. Virology 109:174–179
    [Google Scholar]
  34. Zweerink H. J., Courtneidge S. A., Skehel J. J., Crumpton M. J., Askonas B. A. 1977; Cytotoxic T cells kill influenza virus infected cells but do not distinguish between serologically distinct type A viruses. Nature; London: 267354–356
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-71-5-1169
Loading
/content/journal/jgv/10.1099/0022-1317-71-5-1169
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