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Refined methods for propagating vesicular stomatitis virus vectors that are defective for G protein expression

  1. Author:
    Witko, S. E.
    Johnson, J. E.
    Kalyan, N. K.
    Felber, B. K.
    Pavlakis, G. N.
    Sidhu, M. K.
    Hendry, R. M.
    Udem, S. A.
    Parks, C. L.

  2. Author Address

    [Witko, Susan E.; Johnson, J. Erik; Kalyan, Narender K.; Sidhu, Maninder K.; Hendry, R. Michael; Udem, Stephen A.; Parks, Christopher L.] Pfizer Vaccine Res, Pearl River, NY 10965 USA. [Felber, Barbara K.] NCI, Human Retrovirus Pathogenesis Sect, Vaccine Branch, Ctr Canc Res, Frederick, MD 21702 USA. [Pavlakis, George N.] NCI, Human Retrovirus Sect, Vaccine Branch, Ctr Canc Res, Frederick, MD 21702 USA. [Hendry, R. Michael] Ctr Dis Control, Div HIV AIDS Prevent, Atlanta, GA 30329 USA. [Parks, Christopher L.] Int AIDS Vaccine Initiat, Vaccine Design & Dev Lab, Brooklyn, NY 11220 USA.;Kalyan, NK, Pfizer Vaccine Res, 401 N Middletown Rd, Pearl River, NY 10965 USA.;kalyann@wyeth.com
    1. Year: 2010
    2. Date: Mar
  2. Journal: Journal of Virological Methods
    1. 164
    2. 1-2
    3. Pages: 43-50
  4. Type of Article: Article
  5. ISSN: 0166-0934
  1. Abstract:

    Propagation-defective vesicular stomatitis virus (VSV) vectors that encode a truncated G protein (VSV-Gstem) or lack the G gene entirely (VSV-Delta G) are attractive vaccine vectors because they are immunogenic, cannot replicate and spread after vaccination, and do not express many of the epitopes that elicit neutralizing anti-VSV immunity. To consider advancing non-propagating VSV vectors towards clinical assessment, scalable technology that is compliant with human vaccine manufacturing must be developed to produce clinical trial material. Accordingly, two propagation methods were developed for VSV-Gstem and VSV-Delta G vectors encoding HIV gag that have the potential to support large-scale production. One method is based on transient expression of G protein after electroporating plasmid DNA into Vero cells and the second is based on a stable Vero cell line that contains a G gene controlled by a heat shock-inducible transcription unit. Both methods reproducibly supported production of 1 x 10(7) to 1 x 10(8) infectious units (I.U.s) of vaccine vector per milliliter. Results from these studies also showed that optimization of the G gene is necessary for abundant G protein expression from electroporated plasmid DNA or from DNA integrated in the genome of a stable cell line, and that the titers of VSV-Gstem vectors generally exceeded VSV-Delta G. (C) 2009 Elsevier B.V. All rights reserved.

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External Sources

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  2. DOI: 10.1016/j.jviromet.2009.11.023
  3. View record in Web of Science®
  4. WOS: 000276820500008

Library Notes

  1. Fiscal Year: FY2009-2010
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