The usefulness of competitive PCR: Airway gene expression of IL-5, IL-4, IL-4δ2, IL-2, and ifnγ in asthma

E. M. Glare, M. Divjak, M. J. Bailey, E. H. Walters

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7 Citations (Scopus)

Abstract

Background - Asthma has been described as an eosinophilic bronchitis driven by interleukin(IL)-4 and IL-5. The quantification of cytokine mRNA levels in airway samples has been confounded by house-keeping gene expression which differs between and within asthmatics and controls. Methods - The usefulness of competitive reverse transcriptase-polymerase chain reaction (RT-PCR) that is independent of housekeeping gene expression for quantitating the mRNA for interferon (IFN)γ, IL-2, IL-5, IL-4 and its receptor antagonist encoding splicing variant IL-4δ2 was determined in a cross sectional study of 45 normal control subjects and 111 with asthma. Results - Atopic controls and atopic asthmatic subjects expressed more IL-5 than non-atopic controls (p<0.02) in bronchoalveolar lavage (BAL) cells, but not in biopsy specimens. IL-5 mRNA expression in BAL cells from asthmatic subjects using inhaled corticosteroids (ICS) was significantly lower than those not receiving ICS (p=0.04). IL-2 mRNA levels differed with steroid use in biopsy specimens but not in BAL cells. IFNγ, IL-4, and IL-4δ2 mRNA levels did not differ between any groups and were not affected by steroid use. IL-4 and IL-4δ2 mRNA levels were positively correlated (p<0.0001), suggesting coordinated transcription. Conclusions - While the signal differentiation of competitive PCR in asthma may rival that of in situ hybridisation and immunohistochemistry, the method is expensive and wasteful of material.

Original languageEnglish
Pages (from-to)541-548
Number of pages8
JournalThorax
Volume56
Issue number7
DOIs
Publication statusPublished - 9 Jul 2001

Keywords

  • Asthma
  • Competitive PCR
  • Interleukins

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