title: Heterogeneity of pulmonary endothelial cyclic nucleotide response to Pseudomonas aeruginosa ExoY infection.
abstract: Here, we tested the hypothesis that a promiscuous bacterial cyclase synthesizes purine and pyrimidine cyclic nucleotides in the pulmonary endothelium. To test this hypothesis, pulmonary endothelial cells were infected with a strain of the Gram-negative bacterium Pseudomonas aeruginosa that introduces only exoenzyme Y (PA103 ΔexoUexoT::Tc pUCPexoY; ExoY(+)) via a type III secretion system. Purine and pyrimidine cyclic nucleotides were simultaneously detected using mass spectrometry. Pulmonary artery (PAECs) and pulmonary microvascular (PMVECs) endothelial cells both possess basal levels of four different cyclic nucleotides in the following rank order: cAMP > cUMP ≈ cGMP ≈ cCMP. Endothelial gap formation was induced in a time-dependent manner following ExoY(+) intoxication. In PAECs, intercellular gaps formed within 2 h and progressively increased in size up to 6 h, when the experiment was terminated. cGMP concentrations increased within 1 h postinfection, whereas cAMP and cUMP concentrations increased within 3 h, and cCMP concentrations increased within 4 h postinfection. In PMVECs, intercellular gaps did not form until 4 h postinfection. Only cGMP and cUMP concentrations increased at 3 and 6 h postinfection, respectively. PAECs generated higher cyclic nucleotide levels than PMVECs, and the cyclic nucleotide levels increased earlier in response to ExoY(+) intoxication. Heterogeneity of the cyclic nucleotide signature in response to P. aeruginosa infection exists between PAECs and PMVECs, suggesting the intracellular milieu in PAECs is more conducive to cNMP generation.
authors.affiliation: Department of Physiology and Cell Biology, University of South Alabama, Mobile, Alabama; Center for Lung Biology, University of South Alabama, Mobile, Alabama;
authors.forename: K A
authors.initials: KA
authors.lastname: Morrow
authors.affiliation: Institute of Pharmacology, Hannover Medical School, Hannover, Germany;
authors.forename: R
authors.initials: R
authors.lastname: Seifert
authors.affiliation: Research Core Unit Metabolomics, Hannover Medical School, Hannover, Germany;
authors.forename: V
authors.initials: V
authors.lastname: Kaever
authors.affiliation: Department of Pharmacology, University of South Alabama, Mobile, Alabama; Center for Lung Biology, University of South Alabama, Mobile, Alabama;
authors.forename: A L
authors.initials: AL
authors.lastname: Britain
authors.affiliation: Department of Physiology and Cell Biology, University of South Alabama, Mobile, Alabama; Center for Lung Biology, University of South Alabama, Mobile, Alabama;
authors.forename: S L
authors.initials: SL
authors.lastname: Sayner
authors.affiliation: Physician-Scientist Training Program, Department of Medicine, University of Texas-Southwestern Medical Center, Dallas, Texas; Physician-Scientist Training Program, Division of Pulmonary and Critical Care, University of Texas-Southwestern Medical Center, Dallas, Texas;
authors.forename: C D
authors.initials: CD
authors.lastname: Ochoa
authors.affiliation: Department of Pharmacology, University of South Alabama, Mobile, Alabama; Center for Lung Biology, University of South Alabama, Mobile, Alabama;
authors.forename: E A
authors.initials: EA
authors.lastname: Cioffi
authors.affiliation: Department of Medicine, Medical College of Wisconsin, Milwaukee, Wisconsin; and Center for Infectious Disease Research, Medical College of Wisconsin, Milwaukee, Wisconsin.
authors.forename: D W
authors.initials: DW
authors.lastname: Frank
authors.affiliation: Department of Pharmacology, University of South Alabama, Mobile, Alabama; Center for Lung Biology, University of South Alabama, Mobile, Alabama;
authors.forename: T C
authors.initials: TC
authors.lastname: Rich
authors.affiliation: Department of Physiology and Cell Biology, University of South Alabama, Mobile, Alabama; Department of Medicine, University of South Alabama, Mobile, Alabama; Center for Lung Biology, University of South Alabama, Mobile, Alabama; tstevens@southalabama.edu.
authors.forename: T
authors.initials: T
authors.lastname: Stevens
mesh_terms: D002199:Capillary Permeability; D002478:Cells, Cultured; D042783:Endothelial Cells / Q000378:metabolism* / Q000382:microbiology; D054884:Host-Pathogen Interactions; D055806:Microvessels / Q000166:cytology; D009712:Nucleotides, Cyclic / Q000502:physiology*; D011550:Pseudomonas aeruginosa / Q000201:enzymology*; D011651:Pulmonary Artery / Q000166:cytology
chemical_list: D009712:Nucleotides, Cyclic
keywords: compartmentation; microtubule; permeability; pneumonia; second messenger