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

Glare, E. M.; Divjak, M.; Bailey, M. J.; Walters, E. H.
July 2001
Thorax;Jul2001, Vol. 56 Issue 7, p541
Academic Journal
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 housekeeping gene expression which differs between and within asthmatics and controls. Methods--The usefulness of cometitive reverse transcriptase-polymerase chain reaction (RT-PCR) that is indepedent of housekeeping gene expression for quantitating the mRNA for interferon (IFN)Y, IL-2, IL-5, IL-4 and its receptor antagonist encoding splicing variant IL-482 was determined in a cross sectional study of 45 normal control subjects and 111 with asthma. Result-Atopic controls and atopic asthmatic subject expressed more IL-5 than non-atopic controls (p<0.02) in bronchoalveolar 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. IFNy, IL-4, and IL-482 mRNA levels did not differ between any groups and were not differ between any groups and were not affected by steroid use. IL-4 and IL-482 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.


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