Source: Annual Congress 2003 - Molecular biology of chronic lung disease and lung cancer
Disease area: Airway diseases

Abstract

RATIONALE: Accumulation of eosinophils is seen at cholinergic nerves in airways, for example during acute exacerbations of asthma. Our previous work has established that this causes hyperreactivity of nerves due to muscarinic M₂ receptor antagonism by the eosinophil protein, major basic protein (MBP). We have recently established that eosinophils adhere to the cholinergic cell line IMR32 in culture, via interactions between CD11/18 and VLA-4 on eosinophils and ICAM-1 and VCAM-1 respectively on IMR32 cells. The aim of this project is to investigate the effects of eosinophil adhesion on nerve cell function, in particular on cholinergic phenotype.
METHODS: Eosinophils were isolated from blood and co-cultured with IMR32 cells for times varying from 1 to 48 hr. Nerve cell RNA was isolated using Trizol and converted to cDNA by AMV reverse transcriptase. Quantitative PCR for the muscarinic M₂ receptor, cholinergic genes choline acetyltransferase (ChAT), vesicular acetyl choline transferase (VaChT) and acetylcholinesterase (AChE) was carried out using the LightCycler system and yield was normalised by comparison to the ‘housekeeping’ gene b-actin. Nerve cell membrane and cytoplasmic protein were isolated and subjected to western blotting analysis for M₂ and ChAT and VaChT expression respectively.
RESULTS: Eosinophil co-culture with IMR32 cells induced up-regulation of gene and protein expression of M₂, ChAT, VaChT and up-regulation of AChE mRNA. Up-regulation of M₂ was dependent on adhesion via CD11/18 and VLA-4. Up-regulation of M₂ was NFkB dependant.
CONCLUSIONS: Eosinophil adhesion to cholinergic nerve cells alters their cholinergic phenotype.

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Citations should be made in the following way:
N. Durcan, M. T. Walsh, R. Costello (Dublin, Ireland). Eosinophil adhesion alters cholinergic nerve cell phenotype. Eur Respir J 2003; 22: Suppl. 45, 1496

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