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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
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.
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:
- 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.