{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,26]],"date-time":"2026-09-26T04:05:36Z","timestamp":1790395536307,"version":"4.1.0"},"reference-count":76,"publisher":"MDPI AG","issue":"2","license":[{"start":{"date-parts":[[2022,2,18]],"date-time":"2022-02-18T00:00:00Z","timestamp":1645142400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000050","name":"National Heart Lung and Blood Institute","doi-asserted-by":"publisher","award":["HL118334"],"award-info":[{"award-number":["HL118334"]}],"id":[{"id":"10.13039\/100000050","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000050","name":"National Heart Lung and Blood Institute","doi-asserted-by":"publisher","award":["HL118334-S1"],"award-info":[{"award-number":["HL118334-S1"]}],"id":[{"id":"10.13039\/100000050","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Toxins"],"abstract":"<jats:p>The Gram-negative, opportunistic pathogen Pseudomonas aeruginosa utilizes a type III secretion system to inject exoenzyme effectors into a target host cell. Of the four best-studied exoenzymes, ExoU causes rapid cell damage and death. ExoU is a phospholipase A2 (PLA2) that hydrolyses host cell membranes, and P. aeruginosa strains expressing ExoU are associated with poor outcomes in critically ill patients with pneumonia. While the effects of ExoU on lung epithelial and immune cells are well studied, a role for ExoU in disrupting lung endothelial cell function has only recently emerged. Lung endothelial cells maintain a barrier to fluid and protein flux into tissue and airspaces and regulate inflammation. Herein, we describe a pulmonary microvascular endothelial cell (PMVEC) culture infection model to examine the effects of ExoU. Using characterized P. aeruginosa strains and primary clinical isolates, we show that strains expressing ExoU disrupt PMVEC barrier function by causing substantial PMVEC damage and lysis, in a PLA2-dependent manner. In addition, we show that strains expressing ExoU activate the pro-inflammatory caspase-1, in a PLA2-dependent manner. Considering the important roles for mitochondria and oxidative stress in regulating inflammatory responses, we next examined the effects of ExoU on reactive oxygen species production. Infection of PMVECs with P. aeruginosa strains expressing ExoU triggered a robust oxidative stress compared to strains expressing other exoenzyme effectors. We also provide evidence that, intriguingly, ExoU PLA2 activity was detectable in mitochondria and mitochondria-associated membrane fractions isolated from P. aeruginosa-infected PMVECs. Interestingly, ExoU-mediated activation of caspase-1 was partially inhibited by reactive oxygen species scavengers. Together, these data suggest ExoU exerts pleiotropic effects on PMVEC function during P. aeruginosa infection that may inhibit endothelial barrier and inflammatory functions.<\/jats:p>","DOI":"10.3390\/toxins14020152","type":"journal-article","created":{"date-parts":[[2022,2,21]],"date-time":"2022-02-21T08:23:29Z","timestamp":1645431809000},"page":"152","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":21,"title":["ExoU Induces Lung Endothelial Cell Damage and Activates Pro-Inflammatory Caspase-1 during Pseudomonas aeruginosa Infection"],"prefix":"10.3390","volume":"14","author":[{"given":"Kierra S.","family":"Hardy","sequence":"first","affiliation":[{"name":"Department of Microbiology and Immunology, College of Medicine, University of South Alabama, Mobile, AL 36688, USA"},{"name":"Center for Lung Biology, College of Medicine, University of South Alabama, Mobile, AL 36688, USA"},{"name":"Department of Systems Biology, Harvard Medical School, Boston, MA 02115, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2778-8753","authenticated-orcid":false,"given":"Amanda N.","family":"Tuckey","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, College of Medicine, University of South Alabama, Mobile, AL 36688, USA"},{"name":"Center for Lung Biology, College of Medicine, University of South Alabama, Mobile, AL 36688, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0230-7068","authenticated-orcid":false,"given":"Phoibe","family":"Renema","sequence":"additional","affiliation":[{"name":"Center for Lung Biology, College of Medicine, University of South Alabama, Mobile, AL 36688, USA"},{"name":"Department of Physiology and Cell Biology, College of Medicine, University of South Alabama, Mobile, AL 36688, USA"},{"name":"Department of Biomedical Sciences, College of Allied Health, University of South Alabama Mobile, Mobile, AL 36688, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mita","family":"Patel","sequence":"additional","affiliation":[{"name":"Center for Lung Biology, College of Medicine, University of South Alabama, Mobile, AL 36688, USA"},{"name":"Department of Pharmacology, College of Medicine, University of South Alabama, Mobile, AL 36688, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Abu-Bakr","family":"Al-Mehdi","sequence":"additional","affiliation":[{"name":"Center for Lung Biology, College of Medicine, University of South Alabama, Mobile, AL 36688, USA"},{"name":"Department of Pharmacology, College of Medicine, University of South Alabama, Mobile, AL 36688, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Domenico","family":"Spadafora","sequence":"additional","affiliation":[{"name":"Flow Cytometry Core Lab, College of Medicine, University of South Alabama, Mobile, AL 36688, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Cody A.","family":"Schlumpf","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, College of Medicine, University of South Alabama, Mobile, AL 36688, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Robert A.","family":"Barrington","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, College of Medicine, University of South Alabama, Mobile, AL 36688, USA"},{"name":"Center for Lung Biology, College of Medicine, University of South Alabama, Mobile, AL 36688, USA"},{"name":"Flow Cytometry Core Lab, College of Medicine, University of South Alabama, Mobile, AL 36688, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mikhail F.","family":"Alexeyev","sequence":"additional","affiliation":[{"name":"Center for Lung Biology, College of Medicine, University of South Alabama, Mobile, AL 36688, USA"},{"name":"Department of Physiology and Cell Biology, College of Medicine, University of South Alabama, Mobile, AL 36688, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Troy","family":"Stevens","sequence":"additional","affiliation":[{"name":"Center for Lung Biology, College of Medicine, University of South Alabama, Mobile, AL 36688, USA"},{"name":"Department of Physiology and Cell Biology, College of Medicine, University of South Alabama, Mobile, AL 36688, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jean-Francois","family":"Pittet","sequence":"additional","affiliation":[{"name":"Department of Anesthesiology and Perioperative Medicine, Birmingham School of Medicine, University of Alabama, Birmingham, AL 35294, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7889-1526","authenticated-orcid":false,"given":"Brant M.","family":"Wagener","sequence":"additional","affiliation":[{"name":"Department of Anesthesiology and Perioperative Medicine, Birmingham School of Medicine, University of Alabama, Birmingham, AL 35294, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jon D.","family":"Simmons","sequence":"additional","affiliation":[{"name":"Center for Lung Biology, College of Medicine, University of South Alabama, Mobile, AL 36688, USA"},{"name":"Department of Pharmacology, College of Medicine, University of South Alabama, Mobile, AL 36688, USA"},{"name":"Department of Surgery, College of Medicine, University of South Alabama, Mobile, AL 36688, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8244-629X","authenticated-orcid":false,"given":"Diego F.","family":"Alvarez","sequence":"additional","affiliation":[{"name":"Center for Lung Biology, College of Medicine, University of South Alabama, Mobile, AL 36688, USA"},{"name":"Department of Physiology and Cell Biology, College of Medicine, University of South Alabama, Mobile, AL 36688, USA"},{"name":"Department of Physiology and Pharmacology, College of Osteopathic Medicine, Sam Houston State University, Conroe, TX 77304, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jonathon P.","family":"Audia","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, College of Medicine, University of South Alabama, Mobile, AL 36688, USA"},{"name":"Center for Lung Biology, College of Medicine, University of South Alabama, Mobile, AL 36688, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,2,18]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"2659","DOI":"10.1099\/00221287-147-10-2659","article-title":"Prevalence of type III secretion genes in clinical and environmental isolates of Pseudomonas 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