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Related Experiment Videos

Improving the in vitro antigen specific T cell proliferation assay: the use of interferon-alpha to elicit antigen

V von Baehr1, W Mayer, C Liebenthal

  • 1Institute of Medical Immunology, Charité Medical School, Campus Mitte, Humboldt University Berlin, D-10098, Berlin, Germany. baehr@ber.ladr.de

Journal of Immunological Methods
|April 9, 2001
PubMed
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This study enhances the lymphocyte transformation assay (LTT) for T cell immune responsiveness. Modifications improve the stimulation index, aiding diagnosis in immunocompromised patients and detecting low-frequency antigen-specific T cells.

Area of Science:

  • Immunology
  • Cellular immunology
  • T cell assays

Background:

  • The lymphocyte transformation assay (LTT) measures T cell immune responsiveness.
  • Low stimulation index (<3-5) in LTT complicates interpretation of T cell proliferation.
  • Assay limitations hinder diagnosis in immunocompromised individuals and detection of low-frequency antigen-specific T cells.

Purpose of the Study:

  • To improve the sensitivity and reliability of the T cell proliferation assay.
  • To enhance the stimulation index in the lymphocyte transformation assay.
  • To optimize T cell response measurement for clinical diagnostics.

Main Methods:

  • Modified the lymphocyte transformation assay (LTT) by adding interferon-alpha.
  • Utilized fresh autologous serum instead of pooled human AB serum.

Related Experiment Videos

  • Evaluated assay performance with tetanus toxoid, Candida albicans, and TBE viral antigen.

    • Main Results:

    • Significantly enhanced stimulation index with common antigens in sensitized patients.
    • No increase in false positive results observed.
    • Reduced nonspecific background proliferation, particularly between days 4-6.
    • Confirmed positive effects of interferon-alpha in contact allergy studies (nickel, gold).

    Conclusions:

    • The modified proliferation assay significantly increases the signal-to-noise ratio.
    • This enhanced assay is valuable for immunocompromised patients.
    • The improved assay aids in detecting very low frequencies of antigen-specific T cells.