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Related Experiment Videos

Transient modulation and internalization of T4 antigen induced by phorbol esters.

J A Hoxie, D M Matthews, K J Callahan

    Journal of Immunology (Baltimore, Md. : 1950)
    |August 15, 1986
    PubMed
    Summary

    Phorbol esters reduce T4 antigen expression on T cells by inducing internalization, a process requiring cellular metabolism. T cells can recover T4 expression and become resistant to further phorbol ester effects.

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    Area of Science:

    • Immunology
    • Cell Biology
    • Molecular Biology

    Background:

    • Phorbol esters modulate surface antigen and receptor expression in mammalian cells.
    • Specifically, phorbol esters decrease T4 antigen expression on T lymphoblastoid cell lines and peripheral blood T cells.

    Purpose of the Study:

    • To investigate the metabolic requirements for phorbol ester-induced T4 antigen downregulation.
    • To examine the relationship between T4 antigen loss and phorbol ester receptor expression on T cells.

    Main Methods:

    • Used purified peripheral blood T4 cells and T lymphoblastoid cell lines (MOLT-4).
    • Assessed T4 antigen expression and phorbol ester binding sites.
    • Utilized immunofluorescence microscopy to visualize T4 antigen localization.
    • Investigated the effects of temperature and metabolic inhibitors (azide, methylamine, sodium fluoride) on T4 loss.

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    Main Results:

    • T4 antigen loss occurred at low phorbol ester concentrations (10-15% receptor occupancy).
    • T4 loss was inhibited by low temperature (4°C) and metabolic inhibitors, but not DNA synthesis inhibitors.
    • T4 antigen reappeared on cells after prolonged phorbol ester exposure (>2 days).
    • Recovered cells showed resistance to phorbol esters, correlated with reduced receptor binding sites.
    • Immunofluorescence revealed T4 antigen internalization into the cytoplasm.

    Conclusions:

    • Phorbol ester binding to lymphocyte receptors triggers metabolically dependent events leading to T4 antigen internalization.
    • These findings suggest a mechanism for T4 antigen regulation relevant to its role in signal transduction.