Polymerase chain reaction quantification of cytokine messenger RNA expression in peripheral blood mononuclear cells of patients with acute exacerbations of asthma: effect of glucocorticoid therapy

Clin Exp Allergy. 1994 Sep;24(9):854-67. doi: 10.1111/j.1365-2222.1994.tb01808.x.

Abstract

We have measured the expression of messenger ribonucleic acid (RNA) (mRNA) encoding interleukin-5 (IL-5), IL-4, IL-2 and interferon-gamma (IFN gamma) in peripheral blood mononuclear cells (PBMC) from 10 patients with acute exacerbations of asthma and nine non-asthmatic controls. Measurements were repeated in seven of the asthmatics following 7 days of oral glucocorticoid therapy. Total RNA was extracted from the PBMC, reverse transcribed using oligo-(dT) primers and aliquots of the resulting complementary DNA (cDNA) amplified using the polymerase chain reaction (PCR) in the presence of cytokine-specific primers under non-saturating conditions. PCR products were quantified on a relative basis after Southern blotting and probing with radiolabelled internal oligonucleotide probes by computer assisted densitometry of blot autoradiographs. The relative amounts of IL-5 mRNA in PBMC from the asthmatic patients prior to glucocorticoid therapy were greater (P < 0.01) than those in PBMC from non-asthmatic controls. In contrast, there were no differences in the relative amounts of IL-4, IL-2 and IFN gamma mRNA. In the asthmatics, the relative amounts of IL-5 mRNA correlated with the peripheral blood eosinophil counts (P = 0.02). After oral glucocorticoid therapy of the asthmatics, lung function improved and the relative amounts of PBMC IL-5 mRNA were reduced (P = 0.04) and no longer differed from those in PBMC from non-asthmatic controls. Glucocorticoid therapy was not associated with significant changes in the relative amounts of PBMC IL-4, IL-2 and IFN gamma mRNA. PBMC from atopic subjects contained significantly greater quantities of IL-4 mRNA (P = 0.04) but not IL-5, IL-2 and IFN gamma mRNA compared with non-atopic subjects regardless of their asthmatic status. We conclude that PBMC of patients with acute exacerbations of asthma demonstrate elevated expression of mRNA encoding IL-5, but not IL-2, IL-4 and IFN gamma and that the clinical improvement associated with glucocorticoid therapy is associated with a reduction of IL-5 mRNA expression. We further conclude that elevated expression in PBMC of mRNA encoding IL-4 is a feature of atopy but not of asthma. These observations suggest that IL-5 synthesis by activated T-lymphocytes may be relevant to the pathogenesis of asthma, and that inhibition of this release by glucocorticoids may at least partly explain their therapeutic effect in this disease.

Publication types

  • Clinical Trial
  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Actins / genetics
  • Adolescent
  • Adult
  • Aged
  • Aged, 80 and over
  • Asthma / blood*
  • Asthma / drug therapy*
  • Base Sequence
  • Child
  • Cytokines / genetics*
  • DNA, Complementary / analysis
  • DNA, Complementary / genetics
  • Female
  • Glucocorticoids / therapeutic use*
  • Humans
  • Leukocytes, Mononuclear / chemistry
  • Leukocytes, Mononuclear / physiology*
  • Male
  • Middle Aged
  • Molecular Sequence Data
  • Polymerase Chain Reaction / methods
  • RNA, Messenger / blood*
  • Reference Values
  • Reproducibility of Results
  • Sensitivity and Specificity

Substances

  • Actins
  • Cytokines
  • DNA, Complementary
  • Glucocorticoids
  • RNA, Messenger