Heterogeneity of pulmonary endothelial cyclic nucleotide response to pseudomonas aeruginosa ExoY infection

K. A. Morrow, R. Seifert, V. Kaever, A. L. Britain, S. L. Sayner, C. D. Ochoa, E. A. Cioffi, D. W. Frank, T. C. Rich, T. Stevens

Research output: Contribution to journalArticlepeer-review

16 Scopus citations

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 ̴exoUexoT::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.

Original languageEnglish (US)
Pages (from-to)L1199-L1207
JournalAmerican Journal of Physiology - Lung Cellular and Molecular Physiology
Volume309
Issue number10
DOIs
StatePublished - 2015

Keywords

  • Compartmentation
  • Microtubule
  • Permeability
  • Pneumonia
  • Second messenger

ASJC Scopus subject areas

  • Physiology
  • Pulmonary and Respiratory Medicine
  • Physiology (medical)
  • Cell Biology

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