1887

Abstract

A study was conducted to investigate the effects of increasing the amount of each retroviral component on vector production. It was found that, while the components of both amphotropic and ecotropic vectors were expressed independently of each other in a transient transfection system, increasing the amount of the / component resulted in a decrease in virus titres for the amphotropic particles but not ecotropic particles. Analyses of the virus stocks produced indicated that the negative effect on titres was closely linked to the availability of envelope proteins for virion incorporation. The negative effect was not observed for ecotropic particle production in 293T cells, where the ecotropic receptor was absent, but was manifested when production was conducted in 293/12 cells expressing the ecotropic receptor. This suggested that the premature interaction between envelope and receptor in producer cells could limit the amount of envelope available for virion incorporation. In designing optimal vector production systems it is essential, therefore, to balance the concentration of the vector components and to ensure that there is never an excess of Gag/Gag–Pol.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-81-9-2195
2000-09-01
2024-04-20
Loading full text...

Full text loading...

/deliver/fulltext/jgv/81/9/0812195a.html?itemId=/content/journal/jgv/10.1099/0022-1317-81-9-2195&mimeType=html&fmt=ahah

References

  1. Albritton L. M., Tseng L., Scadden D., Cunningham J. M. 1989; A putative murine ecotropic retrovirus receptor gene encodes a multiple membrane-spanning protein and confers susceptibility to virus infection. Cell 57:659–666
    [Google Scholar]
  2. Delwart E. L., Panganiban A. T. 1989; Role of reticuloendotheliosis virus envelope glycoprotein in superinfection interference. Journal of Virology 63:273–280
    [Google Scholar]
  3. Felsenstein K. M., Goff S. P. 1988; Expression of the gag pol fusion protein of Moloney murine leukemia virus without gag protein does not induce virion formation or proteolytic processing. Journal of Virology 62:2179–2182
    [Google Scholar]
  4. Goff S., Traktman P., Baltimore D. 1981; Isolation and properties of Moloney murine leukemia virus mutants: use of a rapid assay for release of virion reverse transcriptase. Journal of Virology 38:239–248
    [Google Scholar]
  5. Hayman M. J. 1978; Viral polyproteins in chick embryo fibroblasts infected with avian sarcoma leukosis viruses. Virology 85:241–252
    [Google Scholar]
  6. Henriksson P., Bosch V. 1998; Inhibition of cellular glycoprotein incorporation into human immunodeficiency virus-like particles by co-expression of additional cellular interaction partner. Virology 251:16–21
    [Google Scholar]
  7. Henriksson P., Pfeiffer T., Zentgraf H., Alke A., Bosch V. 1999; Incorporation of wild-type and C-terminally truncated human epidermal growth factor receptor into human immunodeficiency virus-like particles: insight into the processes governing glycoprotein incorporation into retroviral particles. Journal of Virology 73:9294–9302
    [Google Scholar]
  8. Jamjoom G. A., Naso R. B., Arlonghaus R. B. 1977; Further characterization of intracellular precursor polyproteins of Rauscher leukemia virus. Virology 78:11–34
    [Google Scholar]
  9. Katz R. A., Skalka A. M. 1990; Control of retroviral RNA splicing through maintenance of suboptimal processing signals. Molecular and Cellular Biology 10:696–704
    [Google Scholar]
  10. Katz R. A., Kotler M., Skalka A. M. 1988; cis -Acting intron mutations that affect the efficiency of avian retroviral RNA splicing: implications for mechanisms of control. Journal of Virology 62:2686–2695
    [Google Scholar]
  11. Matano T., Odawara M., Ohshima M., Yoshikura H., Iwamoto A. 1993; trans -Dominant interference with virus infection at two different stages by a mutant envelope protein of Friend murine leukemia virus. Journal of Virology 67:2026–2033
    [Google Scholar]
  12. Odawara T., Oshima M., Doi K., Iwamoto A., Yoshikura H. 1998; Threshold number of provirus copies required per cell for efficient virus production and interference in Moloney murine leukemia virus-infected NIH 3T3 cells. Journal of Virology 72:5414–5424
    [Google Scholar]
  13. Oppermann H., Bishop J. M., Varmus H. E., Levintow L. 1977; A joint product of the genes gag and pol of avian sarcoma virus: a possible precursor of reverse transcriptase. Cell 12:993–1005
    [Google Scholar]
  14. Ragheb J. A., Yu H., Hofmann T., Anderson W. F. 1995; The amphotropic and ecotropic murine leukemia virus envelope TM subunits are equivalent mediators of direct membrane fusion: implications for the role of the ecotropic envelope and receptor in syncytium formation and viral entry. Journal of Virology 69:7205–7215
    [Google Scholar]
  15. Soneoka Y., Cannon P. M., Ramsdale E. E., Griffiths J. C., Romano G., Kingsman S. M., Kingsman A. J. 1995; A transient three-plasmid expression system for the production of high titre retroviral vectors. Nucleic Acids Research 23:628–633
    [Google Scholar]
  16. Sonstegard T. S., Hackett P. B. 1996; Autogenous regulation of RNA translation and packaging by Rous sarcoma virus Pr76gag. Journal of Virology 70:6642–6652
    [Google Scholar]
  17. Swanstrom R., Wills J. W. 1997; Synthesis, assembly, and processing of viral proteins. In Retroviruses pp 263–334 Edited by Coffin J. M., Hughes S. H., Varmus H. E. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  18. Willey R. L., Maldarelli F., Martin M. A., Strebel K. 1992; Human immunodeficiency virus type 1 Vpu protein induces rapid degradation of CD4. Journal of Virology 66:7193–7200
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-81-9-2195
Loading
/content/journal/jgv/10.1099/0022-1317-81-9-2195
Loading

Data & Media loading...

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