Binding of Aspergillus fumigatus spores to lung epithelial cells and basement membrane proteins: relevance to the asthmatic lung.

BACKGROUND

Aspergillus fumigatus is an opportunistic pathogen to which asthmatic subjects are particularly susceptible. The ability of spores of A fumigatus to bind to pulmonary cells and basement membrane proteins was investigated to determine the mechanisms involved in this susceptibility.

METHODS

Cells of the A549 pulmonary epithelial cell line or purified basement membrane proteins were immobilised on the wells of microtitre plates. They were then exposed to spores of A fumigatus in suspension, with or without various pretreatments of the spores, cells, and proteins. Adherent spores were counted by light microscopy.

RESULTS

Spores of A fumigatus bound in a concentration dependent manner to A549 epithelial cells and pretreatment of cells with interferon gamma (2500 units/ml) caused a significant doubling of spore binding. Binding of spores to A549 cells was inhibited by about a third by pre-incubation of the spores with fibrinogen (100 micrograms/ml). Spores bound specifically to extracellular matrix (ECM) components laid down by A549 cells, and pretreatment of the ECM components with hydrogen peroxide (25-80 microM) enhanced spore binding by approximately one third. They also bound specifically and in a saturable manner to purified fibrinogen, fibronectin, laminin, type I collagen, and type IV collagen. Pre-incubation of spores with Arg-Gly-Asp tripeptide (RGD; 50-200 micrograms/ ml) inhibited binding to fibronectin and type I collagen by 50%.

CONCLUSIONS

This study suggests that the presence of activated epithelial cells and the exposure of basement membrane that occurs in asthma, together with oxidant stress, may facilitate the colonisation of the asthmatic lung by A fumigatus. The RGD sequence may be involved in spore binding to some ECM proteins. Free fibrinogen may protect against binding of A fumigatus spores to the pulmonary epithelium.