Pseudomonas aeruginosa-induced production of free radicals by IFNgamma plus TNFalpha-activated human endothelial cells: mechanism of host defense or of bacterial pathogenesis?
We have previously shown that human umbilical vein endothelial cells (HUVEC) can be activated by IFNgamma plus TNFalpha to kill intracellular (IC) Pseudomonas aeruginosa through production of reactive oxygen intermediate, but the cumulative effects of cytokine activation and bacterial infection on host cells has not been extensively addressed. In this study we investigated the fate of IFNgamma plus TNFalpha-activated HUVEC that have harboured IC bacteria for up to 24 h. At 10 h, the endothelial cell killing of P. aeruginosa isolates exceeded 90%. IC bacteria enhanced the expression of inducible nitric oxide synthase (iNOS) and induced overproduction of NO and superoxide by infected HUVEC. P. aeruginosa IC infection also induced a slight decrease in the cellular level of reduced glutathione (GSH). Overproduction of NO correlated with a marked peroxidation of plasma membrane lipids and decline in HUVEC viability. Treatment of cells with the antioxidant alpha-lipoic acid significantly increased the survival of infected cells. Our data suggest that with the failure of adequate scavenger mechanisms, oxidant radicals overproduced in response to bacterial infection were highly toxic to host cells. Therefore, instead of contributing to defence against infectious agents, the upregulation of free radicals production by endothelial cells in response to cytokine activation would be detrimental to the host.
Department of Microbiology and Immunology, State University of Rio de Janeiro, Brazil.
Maria-Cristina
MC
De Assis
Alessandra M
AM
Saliba
Luciana A
LA
Vidipó
João B
JB
De Salles
Maria-Cristina
MC
Plotkowski
D042783:Endothelial Cells / Q000276:immunology / Q000382:microbiology / Q000473:pathology; D004730:Endothelium, Vascular / Q000276:immunology* / Q000382:microbiology*; D005260:Female; D006801:Humans; D007371:Interferon-gamma / Q000494:pharmacology*; D015227:Lipid Peroxidation; D008563:Membrane Lipids / Q000378:metabolism; D009569:Nitric Oxide / Q000096:biosynthesis*; D011247:Pregnancy; D011552:Pseudomonas Infections / Q000276:immunology / Q000382:microbiology; D011550:Pseudomonas aeruginosa / Q000472:pathogenicity*; D017382:Reactive Oxygen Species / Q000378:metabolism; D013481:Superoxides / Q000378:metabolism*; D008063:Thioctic Acid / Q000494:pharmacology; D013997:Time Factors; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology*
D008563:Membrane Lipids; D017382:Reactive Oxygen Species; D014409:Tumor Necrosis Factor-alpha; D013481:Superoxides; D009569:Nitric Oxide; D008063:Thioctic Acid; D007371:Interferon-gamma