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Transepithelial Bacterial-Induced Neutrophil Are Used To Coordinate 4 Leukotriene B and 3 Distinct Cellular Sources of Hepoxilin A

Neutrophilic inﬁltration is a leading contributor to pathology in a number of pulmonary disease states, including cystic ﬁbrosis. Hepoxilin A 3 (HXA 3 ) is a chemotactic eicosanoid shown to mediate the transepithelial passage of neutrophils in response to infection in several model systems and at multiple mucosal surfaces. Another well-known eicosanoid mediating general neutrophil chemotaxis is leukotriene B 4 (LTB 4 ). We sought to distinguish the roles of each eicosanoid in the context of infection of lung epithelial monolayers by Pseudomonas aeruginosa . Using human and mouse in vitro transwell model systems, we used a combination of biosynthetic inhibitors, receptor antagonists, as well as mutant sources of neutrophils to assess the contribution of each chemoattractant in driving neutrophil transepithelial migration. We found that following chemotaxis to epithelial-derived HXA 3 signals, neutrophil-derived LTB 4 is required to amplify the magnitude of neutrophil migration. LTB 4 signaling is not required for migration to HXA 3 signals, but LTB 4 generation by migrated neutrophils plays a signiﬁcant role in augmenting the initial HXA 3 mediated migration. We conclude that HXA 3 and LTB 4 serve independent roles to collectively coordinate an effective neutrophilic transepithelial migratory response. The Journal of Immunology , 2015, 194: 000–000.