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Related Experiment Videos

Interferon and phorbol esters down-regulate sIgM expression by independent pathways.

F M Schaffer1, S H Benedict, D Petsche

  • 1Division of Immunology, Hospital for Sick Children, Toronto, Canada.

Journal of Cellular Physiology
|February 1, 1988
PubMed
Summary
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Recombinant interferon alpha 2 (IFN-alpha 2) and phorbol esters like TPA significantly reduce surface IgM (sIgM) expression on Daudi cells. These agents act through independent pathways, affecting mRNA levels and enzyme activity differently.

Area of Science:

  • Immunology
  • Molecular Biology
  • Cell Biology

Background:

  • Surface immunoglobulin M (sIgM) expression is a key marker on B cells.
  • Interferons and phorbol esters are known to modulate cellular functions.
  • Daudi cells are a human B lymphoblastoid cell line often used in immunological studies.

Purpose of the Study:

  • To investigate the effects of recombinant interferon alpha 2 (IFN-alpha 2) and phorbol esters (TPA, PDB) on sIgM expression in Daudi cells.
  • To elucidate the molecular mechanisms underlying the down-regulation of sIgM.
  • To determine if the pathways for sIgM down-regulation by IFN-alpha 2 and phorbol esters are independent.

Main Methods:

  • Flow cytometry was used to quantify sIgM expression via relative fluorescence index (RFI).

Related Experiment Videos

  • Cell proliferation and cell cycle progression were monitored.
  • Quantitative molecular techniques assessed mRNA levels for the IgM heavy chain (c mu).
  • Enzyme activity assays measured 2',5'-oligoadenylate (2-5A) synthetase activity.

    • Main Results:

    • IFN-alpha 2 decreased sIgM expression by 60% without affecting proliferation.
    • TPA and PDB decreased sIgM expression by 67% while enhancing proliferation.
    • Combined IFN-alpha 2 and TPA treatment resulted in a >75% decrease in sIgM expression.
    • Both IFN-alpha 2 and TPA reduced steady-state mRNA levels of c mu, indicating pretranslational down-regulation.
    • IFN-alpha 2 increased 2-5A synthetase activity, whereas TPA decreased it when combined with IFN-alpha 2.

    Conclusions:

    • IFN-alpha 2 and phorbol esters independently down-regulate sIgM expression at the pretranslational level in Daudi cells.
    • The agents exhibit additive effects on sIgM down-regulation but opposing effects on cellular processes like proliferation and 2-5A synthetase activity.
    • These findings suggest distinct molecular pathways are involved in the modulation of sIgM by interferons and phorbol esters.