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Related Experiment Videos

Vesicular stomatitis virus phenotypically mixed with retroviruses: an efficient detection method.

J Závada, G Russ, Z Závadová

    Acta Virologica
    |March 1, 1983
    PubMed
    Summary
    This summary is machine-generated.

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    Two methods for assaying vesicular stomatitis virus (VSV) pseudotypes revealed significant differences in detecting retroviral antigens. Immunoprecipitation identified nearly 100% of VSV particles containing retroviral antigens, unlike neutralization assays.

    Area of Science:

    • Virology
    • Molecular Biology
    • Immunology

    Background:

    • Phenotypic mixing allows viruses to acquire surface antigens from other viruses.
    • Vesicular stomatitis virus (VSV) is frequently used as a model system for studying viral particle assembly and entry.
    • Retroviruses, such as Moloney murine leukemia virus (MuLV), express surface antigens crucial for their life cycle.

    Purpose of the Study:

    • To compare the efficacy of two distinct assay methods for detecting phenotypically mixed VSV particles.
    • To quantify the proportion of VSV virions containing retroviral antigens using different detection techniques.
    • To investigate the discrepancies in pseudotype detection between neutralization and immunoprecipitation assays.

    Main Methods:

    • Phenotypic mixing of VSV with retrovirus (MuLV)-coded antigens in cell cultures (SIRC and L cells).

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  • Assaying for phenotypically mixed particles using neutralization assays with specific antisera (anti-VSV and anti-MuLV).
  • Employing immunoprecipitation with specific antibodies and Staphylococcus aureus for detecting viral antigens on VSV particles.

    • Main Results:

    • Neutralization assays detected a low proportion (10^-4) of VSV pseudotypes resistant to anti-VSV and neutralized by anti-MuLV serum.
    • VSV produced in mouse L cells showed minimal pseudotype activity in neutralization tests.
    • Immunoprecipitation revealed that nearly 100% of VSV virions produced in both cell types contained MuLV-related antigen molecules.

    Conclusions:

    • Immunoprecipitation is a more sensitive method for detecting retroviral antigens on VSV particles compared to neutralization assays.
    • Phenotypic mixing results in a high frequency of VSV particles carrying retroviral antigens, even if not all exhibit functional pseudotype activity.
    • The choice of assay method significantly impacts the perceived frequency of viral pseudotypes and antigen incorporation.