title: Basolateral invasion and trafficking of Campylobacter jejuni in polarized epithelial cells.
abstract: Campylobacter jejuni is a major cause of bacterial diarrheal disease. Most enteropathogenic bacteria including C. jejuni can invade cultured eukaryotic cells via an actin- and/or microtubule-dependent and an energy-consuming uptake process. Recently, we identified a novel highly efficient C. jejuni invasion pathway that involves bacterial migration into the subcellular space of non-polarized epithelial cells (termed subvasion) followed by invasion from the cell basis. Here we report cellular requirements of this entry mechanism and the subsequent intracellular trafficking route of C. jejuni in polarized islands of Caco-2 intestinal epithelial cells. Advanced microscopy on infected cells revealed that C. jejuni invades the polarized intestinal cells via the subcellular invasion pathway. Remarkably, invasion was not blocked by the inhibitors of microtubule dynamics colchicine or paclitaxel, and was even enhanced after disruption of host cell actin filaments by cytochalasin D. Invasion also continued after dinitrophenol-induced cellular depletion of ATP, whereas this compound effectively inhibited the uptake of invasive Escherichia coli. Confocal microscopy demonstrated that intracellular C. jejuni resided in membrane-bound CD63-positive cellular compartments for up to 24 h. Establishment of a novel luciferase reporter-based bacterial viability assay, developed to overcome the limitations of the classical bacterial recovery assay, demonstrated that a subset of C. jejuni survived intracellularly for up to 48 h. Taken together, our results indicate that C. jejuni is able to actively invade polarized intestinal epithelial cells via a novel actin- and microtubule-independent mechanism and remains metabolically active in the intracellular niche for up to 48 hours.
authors: Department of Infectious Diseases and Immunology, Utrecht University, Utrecht, The Netherlands.
authors: Lieneke I
authors: LI
authors: Bouwman
authors: Paula
authors: P
authors: Niewold
authors: Jos P M
authors: JP
authors: van Putten
mesh_terms: D000199:Actins / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000378:metabolism; D002169:Campylobacter Infections; D016123:Campylobacter jejuni / Q000472:pathogenicity*; D002460:Cell Line; D003967:Diarrhea; D004847:Epithelial Cells / Q000378:metabolism / Q000382:microbiology*; D006801:Humans; D007413:Intestinal Mucosa / Q000378:metabolism / Q000382:microbiology*; D008247:Lysosomes / Q000378:metabolism / Q000382:microbiology; D008870:Microtubules / Q000378:metabolism
chemical_list: D000199:Actins; D000255:Adenosine Triphosphate
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