Comparative characterization of transfection-and infection-derived simian immunodeficiency virus challenge stocks for In Vivo nonhuman primate studies

Gregory Q. Del Prete, Matthew Scarlotta, Laura Newman, Carolyn Reid, Laura M. Parodi, James D. Roser, Kelli Oswald, Preston A. Marx, Christopher J. Miller, Ronald C. Desrosiers, Dan H. Barouch, Ranajit Pal, Michael Piatak, Elena Chertova, Luis D. Giavedoni, David H. O'Connor, Jeffrey D. Lifson, Brandon F. Keele

Research output: Contribution to journalArticle

51 Scopus citations

Abstract

Simian immunodeficiency virus (SIV) stocks for in vivo nonhuman primate models of AIDS are typically generated by transfection of 293T cells with molecularly cloned viral genomes or by expansion in productively infected T cells. Although titers of stocks are determined for infectivity in vitro prior to in vivo inoculation, virus production methods may differentially affect stock features that are not routinely analyzed but may impact in vivo infectivity, mucosal transmissibility, and early infection events. We performed a detailed analysis of nine SIV stocks, comprising five infection-derived SIVmac251 viral swarm stocks and paired infection- and transfected-293T-cell-derived stocks of both SIVmac239 and SIVmac766. Representative stocks were evaluated for (i) virus content, (ii) infectious titer, (iii) sequence diversity and polymorphism frequency by single-genome amplification and 454 pyrosequencing, (iv) virion-associated Env content, and (v) cytokine and chemokine content by 36-plex Luminex analysis. Regardless of production method, all stocks had comparable particle/infectivity ratios, with the transfected-293T stocks possessing the highest overall virus content and infectivity titers despite containing markedly lower levels of virion-associated Env than infection-derived viruses. Transfected-293T stocks also contained fewer and lower levels of cytokines and chemokines than infection-derived stocks, which had elevated levels of multiple analytes, with substantial variability among stocks. Sequencing of the infection-derived SIVmac251 stocks revealed variable levels of viral diversity between stocks, with evidence of stock-specific selection and expansion of unique viral lineages. These analyses suggest that there may be underappreciated features of SIV in vivo challenge stocks with the potential to impact early infection events, which may merit consideration when selecting virus stocks for in vivo studies.

Original languageEnglish (US)
Pages (from-to)4584-4595
Number of pages12
JournalJournal of virology
Volume87
Issue number8
DOIs
StatePublished - Apr 1 2013
Externally publishedYes

ASJC Scopus subject areas

  • Microbiology
  • Immunology
  • Insect Science
  • Virology

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  • Cite this

    Del Prete, G. Q., Scarlotta, M., Newman, L., Reid, C., Parodi, L. M., Roser, J. D., Oswald, K., Marx, P. A., Miller, C. J., Desrosiers, R. C., Barouch, D. H., Pal, R., Piatak, M., Chertova, E., Giavedoni, L. D., O'Connor, D. H., Lifson, J. D., & Keele, B. F. (2013). Comparative characterization of transfection-and infection-derived simian immunodeficiency virus challenge stocks for In Vivo nonhuman primate studies. Journal of virology, 87(8), 4584-4595. https://doi.org/10.1128/JVI.03507-12