The usefulness of competitive PCR: airway gene expression of IL-5, IL-4, IL-4[delta]2, IL-2, and IFN[gamma] in asthma. (Statistical Data Included)
Glare, EM and Divjak, M and Bailey, MJ and Walters, EH, The usefulness of competitive PCR: airway gene expression of IL-5, IL-4, IL-4[delta]2, IL-2, and IFN[gamma] in asthma. (Statistical Data Included), Thorax, 56, (7) pp. 541-548. ISSN 0040-6376 (2001) [Refereed Article]
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.