Increased interleukin-13 mRNA expression in bronchoalveolar lavage cells of atopic patients with mild asthma after repeated low-dose allergen provocations

Abstract

Immune and inflammatory responses mediated by cytokines are essential in the pathophysiology of asthma. The aim of this study was to analyse the cytokine mRNA profiles in bronchoalveolar lavage (BAL) cells of patients with mild atopic asthma, before and after induction of a subclinical allergic airway inflammation. For this purpose, eight patients with mild atopic asthma received low-dose allergen inhalations equivalent to 10% of a provocational dose causing a 20% fall in forced expiratory flow in 1 sec (PD20) for 7 weekdays. BAL was performed before and after low-dose provocations in patients, and without provocation in five healthy controls. Alveolar macrophages (AM) were enriched by negative selection, using magnetic beads, to enable separate studies of the BAL cells. Using a semiquantitative RT-PCR technique, the mRNA expression of macrophage-derived cytokines interleukin (IL)-1, IL-6, IL-8, IL-10, IL-12, IL-13, interferon (IFN)- γ, tumour necrosis factor (TNF)- α and transforming growth factor (TGF)- β was analysed. After low-dose provocations, we observed a significant increase in the expression of IL-13 mRNA (P=0·01) in BAL cells enriched for AM of the asthmatic patients. The increased IL-13 mRNA positively correlated with the proportion of BAL fluid eosinophils (r=0·7, P=0·05). Moreover, a tendency was found towards an increased IL-1 and a reduced IL-6, IL-8, IFN- γ and TNF- α expression by the BAL cells. Comparing asthmatic patients before low-dose provocations and healthy controls, a significantly higher expression of IL-6 (P<0·003), IL-10 (P<0·005) and TGF- β (P<0·003) and a significantly lower expression of IL-8 (P<0·005) and TNF- α (P<0·01) was detected in the patients. In summary, repeated low-dose allergen provocations of asthmatic patients results in a modified BAL cell cytokine mRNA profile with increased production of IL-13, that may be of importance for the development of a Th2-like immune response. A possible source of the increased IL-13 mRNA is AM, which may have a more active function in the allergic inflammation than previously thought.