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Related Experiment Videos

Mitogenic activation of EL-4 cells does not require surface Thy-1 expression.

K C Gunter1, R N Germain, O Leo

  • 1Laboratory of Immunology, National Institute of Allergy and Infectious Diseases, Bethesda, Maryland 20892.

Cellular Immunology
|March 1, 1988
PubMed
Summary

Thy-1 expression is not essential for T-cell activation. Mutagenesis of T lymphoma cells unexpectedly created non-responsive variants, indicating other mutations, not Thy-1 loss, caused this defect.

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Area of Science:

  • Immunology
  • Molecular Biology
  • Cell Biology

Background:

  • Monoclonal anti-Thy-1 antibodies stimulate IL-2 production and T-cell proliferation.
  • This suggests a potential role for Thy-1 in T-cell activation.

Purpose of the Study:

  • To investigate the role of Thy-1 in T-cell activation.
  • To produce and characterize Thy-1-negative T lymphoma cell lines.

Main Methods:

  • Generated Thy-1-negative variants of murine T lymphoma EL-4 using ethyl methanesulfonate (EMS) mutagenesis and negative selection.
  • Assessed IL-2 production in response to various stimuli (Con A, PHA, anti-Thy-1, anti-T3, PMA).
  • Reconstituted Thy-1 expression via gene transfection and 5-azadeoxycytidine treatment.

Main Results:

Related Experiment Videos

  • Thy-1-negative cell lines produced IL-2 only in response to PMA, unlike parental cells.
  • Loss of responsiveness correlated with Thy-1 loss, but reconstitution did not restore response to mitogens.
  • Independently derived Thy-1-negative lines responded normally to mitogens.

Conclusions:

  • Thy-1 expression is not required for T-cell activation.
  • EMS mutagenesis likely introduced secondary mutations affecting T-cell activation pathways.
  • Thy-1-negative, mitogen-unresponsive cell lines are valuable tools for studying T-cell activation.