title: Are free radicals responsible for endothelial cell killing of Staphylococcus aureus?
abstract: We have recently demonstrated that endothelial cells cultured on Gelfoam blocks, but not monolayer matrices can phagocytose and kill Staphylococcus aureus. Experiments determined that penicillin G, included in the endothelial cell growth medium, induces these cells to exhibit the observed bactericidal activity. In this communication, we report on studies aimed at elucidating the mechanism by which penicillin G-induced endothelial cells, cultured on Gelfoam blocks, kill S. aureus. Despite the fact that there is a substantial literature that demonstrates neutrophilic killing of bacteria can be mediated through free radical-dependent and free radical-independent mechanisms, considerably less is known about pathways by which endothelial cells can catalyze similar microbicidal activities. Studies described herein point to the fact that superoxide and products derived from this free radical were not responsible for endothelial killing of S. aureus. Likewise, a possible role for nitric oxide in bacterial killing was explored. As part of this inquiry, we stably transduced a NOS-2 encoding retrovirus into endothelial cells cultured on Gelfoam blocks in the absence of penicillin G. Even though these cells secreted nitric oxide at a rate of 0.5 microM/h per 1 x 10(6) cells, similar to what has been reported for murine macrophages induced with gamma-interferon, in our model, nitric oxide was not found to kill S. aureus. Data presented demonstrate that the microbicidal activity of endothelial cells is mediated through free radical-independent pathways.
authors.affiliation: Department of Pharmaceutical Sciences, Program in Pharmacology and Toxicology, University of Maryland School of Pharmacy, Baltimore 21201, USA.
authors.forename: B
authors.initials: B
authors.lastname: Zhang
authors.forename: M
authors.initials: M
authors.lastname: Centra
authors.forename: G L
authors.initials: GL
authors.lastname: Cao
authors.forename: R E
authors.initials: RE
authors.lastname: Ratych
authors.forename: J B
authors.initials: JB
authors.lastname: Domachowske
authors.forename: H L
authors.initials: HL
authors.lastname: Malech
authors.forename: G M
authors.initials: GM
authors.lastname: Rosen
mesh_terms: D000818:Animals; D002374:Catalase / Q000494:pharmacology; D002417:Cattle; D002478:Cells, Cultured; D004730:Endothelium, Vascular / Q000187:drug effects / Q000382:microbiology / Q000502:physiology*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005609:Free Radicals; D005781:Gelatin Sponge, Absorbable; D007527:Isoenzymes / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D051379:Mice; D017942:Nitric Acid / Q000378:metabolism; D019001:Nitric Oxide Synthase / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D009573:Nitrites / Q000032:analysis; D010400:Penicillin G / Q000494:pharmacology; D010957:Plasmids; D011993:Recombinant Fusion Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism; D013211:Staphylococcus aureus; D013481:Superoxides / Q000378:metabolism*; D014162:Transfection; D019323:omega-N-Methylarginine / Q000494:pharmacology
chemical_list: D004791:Enzyme Inhibitors; D005609:Free Radicals; D007527:Isoenzymes; D009573:Nitrites; D011993:Recombinant Fusion Proteins; D013481:Superoxides; D019323:omega-N-Methylarginine; D017942:Nitric Acid; D002374:Catalase; D019001:Nitric Oxide Synthase; D010400:Penicillin G