Skip to main content
PMC home page
Journal of Virology logoLink to Journal of Virology
. 1981 Jul;39(1):325–329. doi: 10.1128/jvi.39.1.325-329.1981

Isolation of monoclonal antibodies against avian oncornaviral protein p19.

I Greiser-Wilke, K M Owada, K Moelling
PMCID: PMC171295  PMID: 6268819

Abstract

For the production of monoclonal antibodies against pp60src and the gag precursor protein Pr76gag, the spleens of mice bearing tumors that had been induced by avian sarcoma virus Schmidt-Ruppin D-transformed cells were used. One hybridoma culture produced antibodies that were directed against the p19 portion of the gag precursor. However, no antibodies directed against pp60src could be detected in any of the hybridoma supernatants. The anti-p19-producing hybridoma culture was cloned twice in soft agar, and a stable clone was used for the production of high-titer ascites fluid in mice. The monoclonal antibodies belonged to the immunoglobulin G subclass 2b. The antibodies precipitated Pr76gag and the processed virion-associated p19, as well as the 75,000-molecular-weight gag fusion protein from avian erythroblastosis virus-transformed bone marrow cells. Also, viral ribonucleoprotein complexes were specifically precipitable, indicating that they contain p19 molecules.

Full text

PDF
328

Images in this article

326Image
on p.326
327Image
on p.327
328Image
on p.328

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Baltimore D. Tumor viruses: 1974. Cold Spring Harb Symp Quant Biol. 1975;39(Pt 2):1187–1200. doi: 10.1101/sqb.1974.039.01.137. [DOI] [PubMed] [Google Scholar]
  2. Beug H., Kitchener G., Doederlein G., Graf T., Hayman M. J. Mutant of avian erythroblastosis virus defective for erythroblast transformation: deletion in the erb portion of p75 suggests function of the protein in leukemogenesis. Proc Natl Acad Sci U S A. 1980 Nov;77(11):6683–6686. doi: 10.1073/pnas.77.11.6683. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Brugge J. S., Erikson R. L. Identification of a transformation-specific antigen induced by an avian sarcoma virus. Nature. 1977 Sep 22;269(5626):346–348. doi: 10.1038/269346a0. [DOI] [PubMed] [Google Scholar]
  4. Chen M., Garon C. F., Papas T. S. Native ribonucleoprotein is an efficient transcriptional complex of avian myeloblastosis virus. Proc Natl Acad Sci U S A. 1980 Mar;77(3):1296–1300. doi: 10.1073/pnas.77.3.1296. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Collett M. S., Erikson R. L. Protein kinase activity associated with the avian sarcoma virus src gene product. Proc Natl Acad Sci U S A. 1978 Apr;75(4):2021–2024. doi: 10.1073/pnas.75.4.2021. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Eisenman R., Vogt V. M., Diggelmann H. Synthesis of avian RNA tumor virus structural proteins. Cold Spring Harb Symp Quant Biol. 1975;39(Pt 2):1067–1075. doi: 10.1101/sqb.1974.039.01.122. [DOI] [PubMed] [Google Scholar]
  7. Hayman M. J., Royer-Pokora B., Graf T. Defectiveness of avian erythroblastosis virus: synthesis of a 75K gag-related protein. Virology. 1979 Jan 15;92(1):31–45. doi: 10.1016/0042-6822(79)90212-5. [DOI] [PubMed] [Google Scholar]
  8. Kennett R. H. Cell fusion. Methods Enzymol. 1979;58:345–359. doi: 10.1016/s0076-6879(79)58149-x. [DOI] [PubMed] [Google Scholar]
  9. Köhler G., Milstein C. Continuous cultures of fused cells secreting antibody of predefined specificity. Nature. 1975 Aug 7;256(5517):495–497. doi: 10.1038/256495a0. [DOI] [PubMed] [Google Scholar]
  10. Köhler G., Milstein C. Derivation of specific antibody-producing tissue culture and tumor lines by cell fusion. Eur J Immunol. 1976 Jul;6(7):511–519. doi: 10.1002/eji.1830060713. [DOI] [PubMed] [Google Scholar]
  11. LITTLEFIELD J. W. SELECTION OF HYBRIDS FROM MATINGS OF FIBROBLASTS IN VITRO AND THEIR PRESUMED RECOMBINANTS. Science. 1964 Aug 14;145(3633):709–710. doi: 10.1126/science.145.3633.709. [DOI] [PubMed] [Google Scholar]
  12. Leis J. P., McGinnis J., Green R. W. Rous sarcoma virus p19 binds to specific double-stranded regions of viral RNA: effect of p19 on cleavage of viral RNA by RNase III. Virology. 1978 Jan;84(1):87–98. doi: 10.1016/0042-6822(78)90220-9. [DOI] [PubMed] [Google Scholar]
  13. Moelling K., Scott A., Dittmar K. E., Owada M. Effect of p15-associated protease from an avian RNA tumor virus on avian virus-specific polyprotein precursors. J Virol. 1980 Feb;33(2):680–688. doi: 10.1128/jvi.33.2.680-688.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Moelling K., Sykora K. W., Dittmar K. E., Scott A., Watson K. F. The isolation of avian viral RNA and polypeptides. J Biol Chem. 1979 May 25;254(10):3738–3742. [PubMed] [Google Scholar]
  15. Nowinski R. C., Lostrom M. E., Tam M. R., Stone M. R., Burnette W. N. The isolation of hybrid cell lines producing monoclonal antibodies against the p15(E) protein of ecotropic murine leukemia viruses. Virology. 1979 Feb;93(1):111–126. doi: 10.1016/0042-6822(79)90280-0. [DOI] [PubMed] [Google Scholar]
  16. Owada M., Donner P., Dittmar K. E., Moelling K. Comparison of protein kinase activities in normal cells and cells transformed by a temperature-sensitive mutant of avian sarcoma virus to those of cell-free viral translational products. Cold Spring Harb Symp Quant Biol. 1980;44(Pt 2):959–965. doi: 10.1101/sqb.1980.044.01.103. [DOI] [PubMed] [Google Scholar]
  17. Owada M., Moelling K. Temperature-sensitive kinase activity associated with various mutants of avian sarcoma viruses which are temperature sensitive for transformation. Virology. 1980 Feb;101(1):157–168. doi: 10.1016/0042-6822(80)90492-4. [DOI] [PubMed] [Google Scholar]
  18. Parsons S. J., Riley S. C., Mullen E. E., Brock E. J., Benjamin D. C., Kuehl W. M., Parsons J. T. Immune response to the src gene product in mice bearing tumors induced by injection of avian sarcoma virus-transformed mouse cells. J Virol. 1979 Oct;32(1):40–46. doi: 10.1128/jvi.32.1.40-46.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Potter M. Immunoglobulin-producing tumors and myeloma proteins of mice. Physiol Rev. 1972 Jul;52(3):631–719. doi: 10.1152/physrev.1972.52.3.631. [DOI] [PubMed] [Google Scholar]
  20. Purchio A. F., Erikson E., Brugge J. S., Erikson R. L. Identification of a polypeptide encoded by the avian sarcoma virus src gene. Proc Natl Acad Sci U S A. 1978 Mar;75(3):1567–1571. doi: 10.1073/pnas.75.3.1567. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Sen A., Todaro G. J. The genome-associated, specific RNA binding proteins of avian and mammalian type C viruses. Cell. 1977 Jan;10(1):91–99. doi: 10.1016/0092-8674(77)90143-x. [DOI] [PubMed] [Google Scholar]
  22. Strand M. Transformation-related antigens identified by monoclonal antibodies. Proc Natl Acad Sci U S A. 1980 Jun;77(6):3234–3238. doi: 10.1073/pnas.77.6.3234. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Williams A. F., Galfrè G., Milstein C. Analysis of cell surfaces by xenogeneic myeloma-hybrid antibodies: differentiation antigens of rat lymphocytes. Cell. 1977 Nov;12(3):663–673. doi: 10.1016/0092-8674(77)90266-5. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Virology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES