title: Interferon-gamma selectively increases epithelial permeability to large molecules by activating different populations of paracellular pores.
abstract: Impairment of the gut epithelial barrier by agents such as IFNgamma may play a key role in the pathogenesis of inflammatory disorders by increasing the paracellular penetration of luminal macromolecules, potentially including bacterial antigens. Owing to limitations of current paracellular probes, little is known about the precise functional changes induced by IFNgamma and how these relate to the development of increased macromolecular permeability. Here we investigate how IFNgamma modulates this pathway in T84 monolayers using a novel profiling technique that resolves different populations of paracellular pores by simultaneous analysis of 24 permeability probes of defined molecular size. Two types of functional pore present in control monolayers, an abundant restrictive pore with a radius of approximately 4.5 Angstrom and a much larger but infrequent, non-restrictive pore, were differentially regulated by IFNgamma. Incubation with IFNgamma dose-dependently and reversibly increased the frequency of the non-restrictive pores while having no significant effect on the restrictive component. Cytokine-induced increases in beta, the descriptor of the non-restrictive pore, correlated closely with increased permeability to large molecules (10 kDa) including E. coli-derived lipopolysaccharide, but not small (0.182 kDa) molecules. This effect was associated with changes in expression of the tight junction proteins occludin and claudin-1. These data suggest that IFNgamma selectively increases the transepithelial flux of large molecules by activating specific pathways within the junctional pore. One hypothesis is that this process may be activated in the early stages of the inflammatory response, facilitating the passage of large and potentially antigenic molecules across the gut without gross disruption of the barrier to small molecules.
authors.affiliation: Gut Barrier Group, Injury Research, Salford Royal Hospitals NHS Trust and Faculty of Medicine and Human Sciences, University of Manchester, Clinical Sciences Building, Hope Hospital, Salford, M6 8HD, UK.
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: Watson
authors.forename: Catherine J
authors.initials: CJ
authors.lastname: Hoare
authors.forename: David R
authors.initials: DR
authors.lastname: Garrod
authors.forename: Gordon L
authors.initials: GL
authors.lastname: Carlson
authors.forename: Geoffrey
authors.initials: G
authors.lastname: Warhurst
mesh_terms: D018938:Caco-2 Cells; D002463:Cell Membrane Permeability / Q000187:drug effects*; D062445:Claudin-1; D062465:Claudin-3; D057167:Claudins; D017097:Electric Impedance; D004705:Endocytosis / Q000187:drug effects; D004847:Epithelial Cells / Q000166:cytology* / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D007371:Interferon-gamma / Q000494:pharmacology*; D008565:Membrane Proteins / Q000378:metabolism; D062793:Occludin; D016062:Porosity / Q000187:drug effects; D011994:Recombinant Proteins; D019108:Tight Junctions / Q000187:drug effects / Q000378:metabolism
chemical_list: C568108:CLDN1 protein, human; C491875:CLDN2 protein, human; C568122:CLDN3 protein, human; D062445:Claudin-1; D062465:Claudin-3; D057167:Claudins; D008565:Membrane Proteins; C568985:OCLN protein, human; D062793:Occludin; D011994:Recombinant Proteins; D007371:Interferon-gamma