title: Superantigen activation of immune cells evokes epithelial (T84) transport and barrier abnormalities via IFN-gamma and TNF alpha: inhibition of increased permeability, but not diminished secretory responses by TGF-beta2.
abstract: Superantigens (SAgs) are extremely potent stimulants of T cell activity that have been implicated in the etiopathophysiology of inflammatory disease. Here, we tested the hypothesis that Staphylococcus aureus enterotoxin B (SEB), a model SAg, can alter epithelial transport and/or barrier functions via immune stimulation. Confluent monolayers of the human colonic T84 epithelial cell line, grown on filter supports, were cocultured with SEB +/- PBMC. Subsequently, T84 transport (consisting of baseline short-circuit current (Isc, indicates net ion transport) and secretory responses to carbachol and forskolin) and barrier functions (consisting of transepithelial resistance and fluxes of 3H-labeled mannitol and 51Cr-EDTA) were examined in Ussing chambers. T84 monolayers cocultured with SEB-activated PBMC displayed a time- and dose-dependent decrease in secretory responses to carbachol and forskolin and a significant increase in permeability. These dramatic changes in epithelial function were not due to reduced epithelial viability. Neutralizing Abs to IFN-gamma partially prevented the transport abnormalities, and Abs to TNF-alpha inhibited the increase in epithelial permeability. Abs to IL-1beta and IL-6 did not modulate the SEB-activated PBMC-induced T84 pathophysiology. Addition of TGF-beta2 to conditioned medium from SEB-activated PBMC partially inhibited the increase in T84 permeability but did not affect the transport abnormalities. We conclude that SAgs can elicit epithelial irregularities characteristic of enteric inflammation and that IFN-gamma and TNF-alpha are key mediators in this coculture model of epithelial dysfunction. Additionally, we would highlight the role that TGF-beta2 may play in preventing prolonged increases in epithelial permeability.
authors.affiliation: Intestinal Disease Research Programme, McMaster University, Hamilton, Ontario, Canada. mckayd@fhs.McMaster.ca
authors.forename: D M
authors.initials: DM
authors.lastname: McKay
authors.forename: P K
authors.initials: PK
authors.lastname: Singh
mesh_terms: D000328:Adult; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000494:pharmacology; D000942:Antigens, Bacterial / Q000276:immunology*; D001692:Biological Transport; D002217:Carbachol / Q000494:pharmacology; D002460:Cell Line; D002463:Cell Membrane Permeability; D018920:Coculture Techniques; D005576:Colforsin / Q000494:pharmacology; D003092:Colitis / Q000378:metabolism; D003106:Colon / Q000166:cytology* / Q000378:metabolism; D017077:Culture Media, Conditioned; D004768:Enterotoxins / Q000276:immunology*; D004847:Epithelial Cells; D004848:Epithelium / Q000378:metabolism; D005260:Female; D006801:Humans; D007371:Interferon-gamma / Q000037:antagonists & inhibitors / Q000276:immunology / Q000502:physiology*; D007408:Intestinal Absorption; D007963:Leukocytes, Mononuclear / Q000276:immunology* / Q000378:metabolism; D008297:Male; D008875:Middle Aged; D011994:Recombinant Proteins / Q000494:pharmacology; D018089:Superantigens / Q000276:immunology*; D016212:Transforming Growth Factor beta / Q000494:pharmacology; D014409:Tumor Necrosis Factor-alpha / Q000037:antagonists & inhibitors / Q000276:immunology / Q000502:physiology*
chemical_list: D000911:Antibodies, Monoclonal; D000942:Antigens, Bacterial; D017077:Culture Media, Conditioned; D004768:Enterotoxins; D011994:Recombinant Proteins; D018089:Superantigens; D016212:Transforming Growth Factor beta; D014409:Tumor Necrosis Factor-alpha; D005576:Colforsin; C031818:enterotoxin B, staphylococcal; D007371:Interferon-gamma; D002217:Carbachol