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Related Experiment Videos

Infectious Sindbis virus transient expression vectors for studying antigen processing and presentation.

C S Hahn1, Y S Hahn, T J Braciale

  • 1Department of Molecular Microbiology, Washington University School of Medicine, St. Louis, MO 63110-1093.

Proceedings of the National Academy of Sciences of the United States of America
|April 1, 1992
PubMed
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A novel Sindbis virus (SIN) vector, dsSIN, enables rapid expression of foreign genes and epitopes. This system effectively sensitizes cells for cytotoxic T lymphocyte (CTL) recognition and primes influenza-specific T-cell responses in vivo.

Area of Science:

  • Virology
  • Molecular Biology
  • Immunology

Background:

  • Sindbis virus (SIN) is a versatile RNA virus.
  • Transient expression systems are crucial for research.

Purpose of the Study:

  • To engineer a Sindbis virus (SIN) vector, dsSIN, for enhanced transient expression.
  • To evaluate the vector's efficacy in expressing foreign proteins and T-cell epitopes.
  • To assess the induction of T-cell responses using the dsSIN system.

Main Methods:

  • Engineering of a dsSIN vector with an additional subgenomic promoter.
  • Construction of dsSIN recombinants expressing chloramphenicol acetyltransferase, truncated influenza hemagglutinin (HA), or HA-derived CTL epitopes.
  • Infection of murine cell lines and mice with dsSIN recombinants.

Related Experiment Videos

  • Assessment of protein expression, target cell sensitization for CTL lysis, and in vivo T-cell priming.

    • Main Results:

    • dsSIN recombinants efficiently expressed high levels of foreign proteins (e.g., chloramphenicol acetyltransferase).
    • Infected cells were effectively sensitized for lysis by major histocompatibility complex (MHC)-restricted cytotoxic T lymphocyte (CTL) clones.
    • Immunization with dsSIN recombinants induced influenza-specific T-cell responses in mice.
    • High-titered recombinant virus stocks were generated rapidly.

    Conclusions:

    • The dsSIN expression system provides a rapid and efficient method for generating recombinant viruses.
    • This system facilitates the study of endogenous antigen processing and presentation of T-cell epitopes.
    • dsSIN is a valuable tool for mapping and analyzing class I MHC-restricted T-cell epitopes.