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Related Experiment Videos

A plaque assay for enumerating antigen-sensitive cells in delayed-type hypersensitivity.

B R Bloom, L Jimenez, P I Marcus

    The Journal of Experimental Medicine
    |July 1, 1970
    PubMed
    Summary

    This study introduces a novel method to count antigen-sensitive lymphocytes by observing RNA virus replication. This technique quantifies immune cells responding to specific antigens, like tuberculin purified protein derivatives (PPD), in delayed hypersensitivity.

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    Area of Science:

    • Immunology
    • Virology
    • Cell Biology

    Background:

    • Resting lymphocytes typically do not support RNA virus replication.
    • Antigen-activated lymphocytes exhibit an increased capacity to support viral replication.
    • Delayed hypersensitivity involves antigen-sensitive lymphocytes.

    Purpose of the Study:

    • To develop and validate a general method for enumerating antigen-sensitive lymphocytes.
    • To quantify lymphocytes responsive to tuberculin purified protein derivatives (PPD) in sensitized guinea pigs.
    • To differentiate antigen-reactive cells from their progeny.

    Main Methods:

    • Lymphocytes from tuberculin-sensitive guinea pigs were cultured with or without PPD.
    • Cells were infected with Newcastle disease virus or vesicular stomatitis virus.

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  • Infectious virus yield was detected via plaque formation in susceptible cell monolayers.

    • Main Results:

    • A significant increase in plaque-forming units (PFU) was observed in PPD-stimulated lymphocytes compared to controls.
    • No increase in PFU was detected in lymphocytes from normal or antibody-producing guinea pigs.
    • The increase in PFU, up to 48 hours, was attributed to initial antigen-reactive cells, not proliferating progeny, confirmed by mitosis inhibitors.

    Conclusions:

    • The described method effectively enumerates antigen-sensitive lymphocytes based on enhanced viral replication.
    • This technique provides a quantitative measure of cellular immune response to specific antigens.
    • The findings highlight the role of initial antigen recognition in delayed hypersensitivity reactions.