The role of primary human alveolar epithelial cells in host defense against Francisella tularensis infection

Department of Microbiology and Immunology, University of Texas Medical Branch, 301 University Blvd., Galveston, Texas 77555-1070; Department of Pediatrics, Sealy Center for Vaccine Development, University of Texas Medical Branch, 301 University Blvd., Galveston, Texas 77555-1436; Department of Pathology, University of Texas Medical Branch, 301 University Blvd., Galveston, Texas 77555-0609

^* To whom correspondence should be addressed. Email: tdeavesp{at}utmb.edu.

	Abstract

F. tularensis (Ft), 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 Ft. Results demonstrated that the live vaccine strain (LVS) contacted ATI and ATII cells by 2h 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 Ft Schu 4 or LVS. Basolateral (BL) conditioned medium (CM), collected 6 and 24 hours later, was added to the BL surface of transwell cultures of primary human pulmonary microvasculature endothelial cells (HPMEC) prior to the addition of neutrophils (PMN) 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 Ft strains induced various levels of a variety of cytokines via NF-kB activation. ATII cells pretreated with an NF-kB inhibitor prior to Ft stimulation substantially decreased IL-8 secretion, which did not occur through Toll-like receptors (TLR) 2, 2/6, 4, or 5 stimulation. These data represent a crucial role for ATII cells in the innate immune response to Ft.