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Updated: Jun 10, 2026

Human In Vitro Suppression as Screening Tool for the Recognition of an Early State of Immune Imbalance
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An in vitro test for immunomodulators?

S Steer1, W Lasek, R H Clothier

  • 1Department of Human Morphology, University of Nottingham Medical School, Queen's Medical Centre, Nottingham NG7 2UH, UK.

Toxicology in Vitro : an International Journal Published in Association with BIBRA
|August 13, 2010
PubMed
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Researchers developed a new assay using interleukin-2 (IL-2) dependent cells to screen chemicals for potential immune effects. This method distinguishes immunomodulatory activity from general cytotoxicity, aiding in the discovery of novel immune-modulating compounds.

Area of Science:

  • Immunology
  • Cell Biology
  • Pharmacology

Background:

  • Interleukin-2 (IL-2) is a critical cytokine mediating immune responses.
  • EL-4 thymoma cells can be stimulated to produce IL-2.
  • IL-2 dependent T-cell lines, like CTLL-2, are crucial for studying IL-2 activity.

Purpose of the Study:

  • To establish and validate a cell-based assay for screening chemicals with potential immunomodulatory effects.
  • To differentiate between direct cytotoxicity and specific immune response modulation by chemical compounds.
  • To assess the utility of the CTLL-2 cell line as a reporter system for IL-2 activity.

Main Methods:

  • Stimulation of EL-4 cells to produce IL-2 using mitogens (phorbol-12-myristate-13-acetate and concanavalin A).
  • Culture of IL-2 dependent CTLL-2 cells in conditioned medium (CM) from stimulated EL-4 cells.

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  • Assessment of CTLL-2 cell response via [(3)H]thymidine incorporation.
  • Determination of ID(50) values for fourteen test chemicals in the CTLL-2 system.
  • Evaluation of basic cytotoxicity using 3T3-L1 fibroblast-like cells.

    • Main Results:

    • The CTLL-2 cell response to CM was measurable via thymidine incorporation.
    • Fourteen chemicals were tested, and their ID(50) values in the CTLL-2 system were determined.
    • Basic cytotoxicity was assessed for the same chemicals using 3T3-L1 cells.
    • Significant discrepancies between CTLL-2 response and basic cytotoxicity were observed for certain chemicals.

    Conclusions:

    • The CTLL-2 responding system shows potential for detecting effects beyond general cytotoxicity.
    • This assay system may be developed to evaluate the immunomodulatory potential of chemicals.
    • Further examination is warranted to fully establish this system as a reliable immunotoxicity screening tool.