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Campylobacter upsaliensis exerts a cytolethal distending toxin effect on HeLa cells and T lymphocytes

Children’s Research Centre, Our Lady’s Hospital for Sick Children and Department of Paediatrics, The Conway Institute of Biomolecular and Biomedical Research, University College Dublin, Crumlin, Dublin 12, Ireland¹

Author for correspondence: B. Bourke. Tel: +353 1 4556901. Fax: +353 1 4555307. e-mail: BILLY.BOURKE{at}ucd.ie

Campylobacter upsaliensis is an emerging human enteropathogen. However, little is known about the pathogenesis of C. upsaliensis infection. In this study the authors demonstrate that C. upsaliensis whole-cell preparations and extracts produce a cytolethal distending toxin (CDT)-like effect on HeLa cells characterized by progressive distension and nuclear fragmentation culminating in cell death over 5 d. To further delineate the nature of this toxic effect in relation to CDT from other pathogens, the effect of C. upsaliensis on cellular events in epithelial cells and immunocytes was investigated. C. upsaliensis lysate-treated HeLa cells subjected to FACScan analysis using carboxyfluorescein diacetete succinimidyl ester (CFDA-SE) as a cell tracer demonstrated cell division arrest. Propidium iodide (PI) staining of HeLa cells revealed that cell cycle arrest occurred in G₂/M. Human T lymphocytes exposed to C. upsaliensis lysates also showed cell cycle arrest in G₂/M. Using a combination of Annexin V/PI staining and TUNEL assay, cytodistended HeLa cells were shown to undergo apoptotic cell death. These data provide the first insights into the virulence mechanisms of this novel enteropathogen.

Keywords: enteropathogen, CDT, virulence factors, apoptosis

Abbreviations: CDT, cytolethal distending toxin; CFDA-SE, carboxyfluorescein diacetete succinimidyl ester; CFSE; carboxyfluorescein succinimidyl ester; PHA, phytohaemagglutinin; PI, propidium iodide

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