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Related Experiment Videos

The active E-rosette test: a sensitive in vitro correlate for human delayed-type hypersensitivity.

P J Felsburg, R Edelman

    Journal of Immunology (Baltimore, Md. : 1950)
    |January 1, 1977
    PubMed
    Summary
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    This study adapted the "active" rosette test for in vitro use, correlating it with human delayed cutaneous hypersensitivity (DCH) to microbial antigens. The new assay shows promise for diagnosing antigen sensitization and studying cell-mediated immunity.

    Area of Science:

    • Immunology
    • Cellular Immunity

    Background:

    • Delayed cutaneous hypersensitivity (DCH) is a key indicator of cell-mediated immunity.
    • Assessing DCH typically requires in vivo skin testing, which can be invasive.
    • There is a need for reliable in vitro methods to evaluate cellular immune responses to microbial antigens.

    Purpose of the Study:

    • To adapt the "active" rosette test as an in vitro assay.
    • To correlate this in vitro assay with human DCH reactions to microbial antigens.
    • To explore its potential as a diagnostic tool for antigen sensitization.

    Main Methods:

    • Peripheral lymphocytes were isolated from individuals with known DCH responses to PPD-tuberculin or tularemia.
    • Lymphocytes were incubated with specific microbial antigens.

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  • The formation of active rosette-forming cells (A-RFC) was measured as an indicator of immune response.

    • Main Results:

    • Antigen-sensitive lymphocytes showed a significant increase in A-RFC formation compared to controls.
    • Lymphocytes from non-sensitive donors did not exhibit increased A-RFC.
    • The assay demonstrated specificity for the sensitizing antigen and an optimal antigen dose of approximately 100 ng/ml.
    • Peak A-RFC levels were observed between 3 and 4 hours of incubation at 37°C.

    Conclusions:

    • The adapted "active" rosette test serves as a valid in vitro correlate of human DCH.
    • This assay offers a potential new method for investigating cell-mediated immunity mechanisms.
    • It may provide a rapid diagnostic test for sensitization to microbial antigens.