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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)

journal contribution
posted on 2023-05-16, 12:50 authored by Glare, EM, Divjak, M, Bailey, MJ, Eugene WaltersEugene Walters
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.

History

Publication title

Thorax

Volume

56

Issue

7

Pagination

541-548

ISSN

0040-6376

Department/School

Tasmanian School of Medicine

Publisher

British Medical Journal Publishing Group

Place of publication

London, England

Repository Status

  • Restricted

Socio-economic Objectives

Clinical health not elsewhere classified

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