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Packaging cell lines for simian foamy virus type 1 vectors.

M Wu1, A Mergia

  • 1Department of Pathobiology, College of Veterinary Medicine, University of Florida, Gainesville, Florida 32610, USA.

Journal of Virology
|April 10, 1999
PubMed
Summary

Stable packaging cell lines for simian foamy virus type 1 (SFV-1) vectors were developed. The inducible packaging cell line produced significantly more vector particles, advancing gene therapy vector production.

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The efficiency of simian foamy virus vector type-1 (SFV-1) in nondividing cells and in human PBLs.

Virology·2001

Area of Science:

  • * Virology and Gene Therapy
  • * Molecular Biology and Vector Development

Background:

  • * Foamy viruses, specifically simian foamy virus type 1 (SFV-1), are nonpathogenic retroviruses with potential for gene transfer.
  • * Previous work established SFV-1 as a useful vector system through transient expression assays.
  • * Development of stable packaging cell lines is crucial for scalable production of SFV-1 based vectors.

Purpose of the Study:

  • * To establish the first stable packaging cell lines for SFV-1 based gene transfer vectors.
  • * To compare the efficiency of SFV-1 vector production using constitutive versus inducible promoters.
  • * To investigate the feasibility of pseudotyping SFV-1 vectors with vesicular stomatitis virus envelope protein G (VSV-G).

Main Methods:

  • * Construction of two SFV-1 packaging cell lines utilizing either a constitutive cytomegalovirus (CMV) immediate-early promoter or an inducible tetracycline promoter.

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  • * Evaluation of vector particle production from both constitutive and inducible packaging cell lines.
  • * Assessment of SFV-1 vector transduction efficiency when pseudotyped with VSV-G.

    • Main Results:

    • * The inducible packaging cell line produced four times more SFV-1 vector particles compared to the constitutive cell line.
    • * Higher copy numbers of packaging DNA in the constitutive cell line did not correlate with increased vector production, suggesting potential toxicity or expression level issues.
    • * SFV-1 vectors pseudotyped with VSV-G showed insignificant transduction levels, indicating poor compatibility for high-titer vector generation.

    Conclusions:

    • * Stable packaging cell lines for SFV-1 vectors have been successfully developed.
    • * Inducible promoters offer advantages for SFV-1 vector production over constitutive promoters.
    • * The development of these stable cell lines is a significant advancement for the scaled-up production of SFV-1 vectors for gene therapy applications.
    • * VSV-G is not suitable for pseudotyping SFV-1 vectors to achieve high transduction titers.