title: Role of primary human alveolar epithelial cells in host defense against Francisella tularensis infection.
abstract: Francisella tularensis, an intracellular pathogen, is highly virulent when inhaled. Alveolar epithelial type I (ATI) and type II (ATII) cells line the majority of the alveolar surface and respond to inhaled pathogenic bacteria via cytokine secretion. We hypothesized that these cells contribute to the lung innate immune response to F. tularensis. Results demonstrated that the live vaccine strain (LVS) contacted ATI and ATII cells by 2 h following intranasal inoculation of mice. In culture, primary human ATI or ATII cells, grown on transwell filters, were stimulated on the apical (AP) surface with virulent F. tularensis Schu 4 or LVS. Basolateral (BL) conditioned medium (CM), collected 6 and 24 h later, was added to the BL surfaces of transwell cultures of primary human pulmonary microvasculature endothelial cells (HPMEC) prior to the addition of polymorphonuclear leukocytes (PMNs) or dendritic cells (DCs) to the AP surface. HPMEC responded to S4- or LVS-stimulated ATII, but not ATI, CM as evidenced by PMN and DC migration. Analysis of the AP and BL ATII CM revealed that both F. tularensis strains induced various levels of a variety of cytokines via NF-kappaB activation. ATII cells pretreated with an NF-kappaB inhibitor prior to F. tularensis stimulation substantially decreased interleukin-8 secretion, which did not occur through Toll-like receptor 2, 2/6, 4, or 5 stimulation. These data indicate a crucial role for ATII cells in the innate immune response to F. tularensis.
authors.affiliation: Department of Microbiology and Immunology, University of Texas Medical Branch, 301 University Blvd., Galveston, TX 77555-1070, USA.
authors.forename: Megan
authors.initials: M
authors.lastname: Gentry
authors.forename: Joanna
authors.initials: J
authors.lastname: Taormina
authors.forename: Richard B
authors.initials: RB
authors.lastname: Pyles
authors.forename: Linsey
authors.initials: L
authors.lastname: Yeager
authors.forename: Michelle
authors.initials: M
authors.lastname: Kirtley
authors.forename: Vsevolod L
authors.initials: VL
authors.lastname: Popov
authors.forename: Gary
authors.initials: G
authors.lastname: Klimpel
authors.forename: Tonyia
authors.initials: T
authors.lastname: Eaves-Pyles
mesh_terms: D000818:Animals; D002465:Cell Movement; D002478:Cells, Cultured; D018925:Chemokines / Q000096:biosynthesis; D017077:Culture Media, Conditioned; D016207:Cytokines / Q000096:biosynthesis; D003713:Dendritic Cells / Q000276:immunology; D004195:Disease Models, Animal; D042783:Endothelial Cells / Q000166:cytology; D004847:Epithelial Cells / Q000166:cytology / Q000276:immunology* / Q000382:microbiology*; D005260:Female; D005604:Francisella tularensis / Q000276:immunology*; D006801:Humans; D007113:Immunity, Innate; D016209:Interleukin-8 / Q000276:immunology; D008168:Lung / Q000382:microbiology / Q000648:ultrastructure; D051379:Mice; D008807:Mice, Inbred BALB C; D046529:Microscopy, Electron, Transmission; D016328:NF-kappa B / Q000378:metabolism; D009504:Neutrophils / Q000276:immunology; D011650:Pulmonary Alveoli / Q000166:cytology / Q000276:immunology* / Q000382:microbiology*; D051193:Toll-Like Receptors / Q000276:immunology; D014406:Tularemia / Q000276:immunology*
chemical_list: D018925:Chemokines; D017077:Culture Media, Conditioned; D016207:Cytokines; D016209:Interleukin-8; D016328:NF-kappa B; D051193:Toll-Like Receptors