title: Heparin-mediated inhibition of Chlamydia psittaci adherence to HeLa cells.
abstract: The adherence of human strains of Chlamydia trachomatis has been recently shown to be inhibitable by heparin and heparitinase, leading to the proposal that Chlamydia binding to host cells may be mediated by a glycosaminoglycan (GAG)-dependent mechanism. We here describe the adherence of the guinea-pig pathogen, Chlamydia psittaci GPIC, to HeLa cells, which was measured by cytofluorometry with chlamydiae whose DNA was fluorescently labelled. Adherence could be inhibited by heat or trypsin pretreatment of the bacteria, and binding was much faster at 37 degrees C (reaching a plateau within 1 h) than 4 degrees C. Little binding remained when host cells were pre-fixed with paraformaldehyde, suggesting that host cell receptor mobility may be required for effective adherence. Visualization by confocal microscopy confirmed that the bacteria were at or near the host cell surface during the entire time-course of these experiments. Adherence increased as a function of pH between pH 6 and pH 8.0-8.5. Both adherence and infection of HeLa cells could be inhibited with heparin when the adherence step was performed at 4 degrees C, but only infection was inhibited when the adherence step was performed at 37 degrees C, even though heparitinase could block adherence at either 4 degrees C or 37 degrees C. Even at 4 degrees C, heparin-mediated inhibition was significantly lower at pH 8 than pH 7.4, suggesting that GAG-independent mechanisms may play a role in the higher adherence observed at basic pH. These results therefore demonstrate that a GAG-dependent adherence step may be operative in C. psittaci, and raise the possibility that other adherence mechanisms may also contribute to binding by this chlamydial strain. Furthermore, they suggest that there may not be a strict correlation between C. psittaci adherence and the ability to cause productive infections.
authors.affiliation: Institut Pasteur, Unité de Biologie des Interactions Cellulaires, CNRS URA, 1960, 75724 Paris.
authors.forename: C B
authors.initials: CB
authors.lastname: Gutiérrez-Martín
authors.forename: D M
authors.initials: DM
authors.lastname: Ojcius
authors.forename: R
authors.initials: R
authors.lastname: Hsia
authors.forename: R
authors.initials: R
authors.lastname: Hellio
authors.forename: P M
authors.initials: PM
authors.lastname: Bavoil
authors.forename: A
authors.initials: A
authors.lastname: Dautry-Varsat
mesh_terms: D001422:Bacterial Adhesion / Q000187:drug effects*; D002478:Cells, Cultured; D002691:Chlamydophila psittaci / Q000254:growth & development / Q000378:metabolism / Q000472:pathogenicity*; D004269:DNA, Bacterial / Q000378:metabolism; D005343:Fibrinolytic Agents / Q000494:pharmacology; D005434:Flow Cytometry; D005557:Formaldehyde / Q000494:pharmacology; D006025:Glycosaminoglycans / Q000378:metabolism; D006367:HeLa Cells; D006361:Heating; D006493:Heparin / Q000494:pharmacology; D006801:Humans; D006863:Hydrogen-Ion Concentration; D018613:Microscopy, Confocal; D011108:Polymers / Q000494:pharmacology; D011133:Polysaccharide-Lyases / Q000494:pharmacology; D009956:Psittacosis / Q000382:microbiology*; D013997:Time Factors; D014357:Trypsin / Q000494:pharmacology
chemical_list: D004269:DNA, Bacterial; D005343:Fibrinolytic Agents; D006025:Glycosaminoglycans; D011108:Polymers; D005557:Formaldehyde; D006493:Heparin; D014357:Trypsin; D011133:Polysaccharide-Lyases; C021349:heparitinsulfate lyase; C003043:paraform