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Methotrexate regulates the expression of glucocorticoid receptor alpha and beta isoforms in normal human peripheral

I Annelise Goecke1, Claudia Alvarez, Juan Henríquez

  • 1Rheumatology Service, Internal Medicine Department, Clinical Hospital, University of Chile, Santos Dumont 999, Santiago, Chile. agoecke@med.uchile.cl

Molecular Immunology
|November 23, 2006
PubMed

Insights

Methotrexate (MTX) increases glucocorticoid receptor alpha (GCR alpha) and decreases GCR beta expression in lymphocytes, enhancing cell sensitivity to steroids. These effects are independent of adenosine receptor A2 signaling.

Area of Science:

  • Immunology
  • Pharmacology
  • Cell Biology

Background:

  • Methotrexate (MTX) is a key therapy for autoimmune-inflammatory diseases, often acting as a steroid-sparing agent.
  • MTX improves glucocorticoid (GC) sensitivity in GC-insensitive conditions like asthma.
  • Glucocorticoid receptor (GCR) signaling is central to GC action, with GCR alpha and GCR beta isoforms playing distinct roles.

Purpose of the Study:

  • To investigate whether MTX regulates GCR expression, specifically altering the balance between GCR alpha and GCR beta.
  • To determine if MTX-induced changes in GCR expression enhance lymphocyte sensitivity to glucocorticoids.
  • To elucidate the role of adenosine and adenosine receptor A2 in mediating MTX's effects on GCR expression.

Main Methods:

  • Quantification of GCR alpha and GCR beta mRNA and protein levels in various human lymphocyte cell lines (CEM, Jurkat, Raji) and peripheral blood mononuclear cells (PBMC) after MTX treatment.
  • Assessment of PBMC sensitivity to dexamethasone (DEX)-induced inhibition of lipopolysaccharide (LPS)-stimulated TNF-alpha release post-MTX treatment.
  • Evaluation of adenosine's direct effect on GCR expression and the impact of an A2 receptor antagonist (DMPX) on MTX-induced GCR modulation.

Main Results:

  • MTX significantly increased GCR alpha mRNA and protein levels in cultured lymphocytes.
  • MTX decreased or did not alter GCR beta protein expression in these cells.
  • MTX-treated PBMCs showed enhanced sensitivity to DEX in inhibiting TNF-alpha release; adenosine did not mimic these effects, and DMPX potentiated MTX's impact on GCRs.

Conclusions:

  • MTX treatment shifts the GCR alpha/GCR beta expression ratio favorably in lymphocytes.
  • This modulation of GCR expression likely underlies MTX's ability to improve cellular glucocorticoid sensitivity.
  • The observed effects of MTX on GCR expression are independent of adenosine receptor A2 signaling.

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