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Role of heparan sulphate proteoglycans as potential receptors for non-piliated Pseudomonas aeruginosa adherence to non-polarised airway epithelial cells

MARIA C. PLOTKOWSKI, ANGELA O. COSTA, VERÔNICA MORANDI^*, HELENE S. BARBOSA, HELENA B. NADER, SOPHIE DE BENTZMANN and EDITH PUCHELLE

Department of Microbiology and Immunology, and *Department of Cell Biology and Genetics, Universidade do Estado do Rio de Janeiro UERJ, Department of Ultrastructure and Cell Biology, FIOCRUZ, Rio de Janeiro, Departament of Biochemistry, Escola Paulista de Medicina, São Paulo, Brazil and U 514 INSERM, IFR 53 Reims, France

Corresponding author: Dr M.C. Plotkowski (e-mail mcplot{at}uerj.br).

Received 3 April 2000; revised version received 30 June 2000; accepted 10 July 2000.

Abstract

Tight junctions seal polarised surface epithelial respiratory cells so as to prevent the passage of bacteria and toxins through the epithelial sheet. Disruption of tight junctions, which may occur during injury and repair processes of airway epithelium, favours potential bacterial interaction with receptors from cell basolateral membranes. Earlier studies reported that non-polarised and untight epithelial respiratory cells are highly susceptible to Pseudomonas aeruginosa adherence and internalisation. As heparan sulphate proteoglycans (HSP) from cell basolateral membranes in epithelial cells without tight junctions may become accessible to bacterial ligands, the present study investigated their role as potential receptors for non-piliate P. aeruginosa ligands. Treatment of cells with heparitinase I and II significantly reduced (51.2% and 51.7%, respectively) P. aeruginosa adherence to epithelial respiratory cells without tight junctions. The internalisation of bacteria was not affected by treatment with heparitinases. Treatment of the bacteria with heparin and heparan sulphate also significantly reduced their adherence to respiratory cells (34.3% and 43.7%, respectively). Treatment of cells with other enzymes (trypsin, lipase and chondroitinase ABC) or treatment of bacteria with chondroitin-4-sulphate did not modify the adherence to respiratory cells significantly. Both affinity chromatography and Western blotting assays showed the interaction of different P. aeruginosa outer-membrane proteins (OMPs) with heparin. Several bacterial strains showed differences in their profile of heparin-binding OMPs, but all exhibited low mol. wt (<5mu305mukDa) reactive proteins. Reactivity of whole bacterial cells with heparin was also observed by transmission electron microscopy. These results suggest that HSP are potential receptors for P. aeruginosa adherence to non-polarised and untight epithelial respiratory cells.

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