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Heterogeneity of Pulmonary Endothelial Cyclic Nucleotide Response to Pseudomonas aeruginosa Infection
Author(s) -
Morrow Kyle,
Seifert Roland,
Britain Andrea,
Ochoa Cristhiaan,
Cioffi Eugene,
Frank Dara,
Rich Thomas,
Stevens Troy
Publication year - 2015
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.29.1_supplement.661.6
Subject(s) - intracellular , extracellular , nucleotide , cyclic nucleotide , multiplicity of infection , biology , endothelial stem cell , microbiology and biotechnology , endothelium , chemistry , in vitro , biochemistry , gene , genetics
Here, we tested the hypothesis that a promiscuous bacterial cyclase generates a diverse cyclic nucleotide response in pulmonary endothelium. Pulmonary endothelial cells were infected with a gram‐negative bacterium Pseudomonas aeruginosa . A strain that introduces only ExoY (PA103 ΔexoUexoT::Tc pUCPexoY; ExoY + ) via a type 3 secretion system was used for cellular infections. Pulmonary artery and pulmonary microvascular endothelial cells (PAECs and PMVECs) were infected with the ExoY + strain at a multiplicity of infection of 20:1. PAECs and PMVECs both possess basal levels of 4 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. Increasing levels of cGMP were seen by 1 h post‐infection, with cAMP and cUMP increasing at 3 h followed by cCMP by 4 h. In PMVECs, intercellular gaps did not form until 4 h with increased levels of only cGMP and cUMP at 3 and 6 h post‐infection, respectively. PAECs generated higher cyclic nucleotide levels than PMVECs, and the cyclic nucleotide levels increased much 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.