title: Auto-protective redox buffering systems in stimulated macrophages.
abstract: BACKGROUND
  Macrophages, upon encounter with micro-organisms or stimulated by cytokines, produce various effector molecules aimed at destroying the foreign agents and protecting the organism. Reactive oxygen species (ROS) and reactive nitrogen species (RNS) are front line molecules exerting strong cytotoxic activities against micro-organisms and many cells, including macrophages themselves. Using cells of the murine macrophage cell line (RAW 264.7) stimulated in vitro with lipopolysaccharide (LPS) and/or interferon (IFN-gamma), which induce strong endogenous NO production, we examined by which mechanisms a fraction of activated macrophages protect themselves from nitrosative stress and manage to escape destruction?
  
  
  RESULTS
  We observed that survivors (10-50% depending on the experiments) had acquired a resistant phenotype being capable to survive when further exposed in vitro to an apoptosis inducing dose of the NO donor compound DETA-NO. These cells expressed an increased steady-state levels of Mn SOD, CuZn SOD and catalase mRNA (130-200%), together with an increased activity of the corresponding enzymes. Intracellular concentration of glutathione was also increased (x 3.5 fold at 6 hours, still maintained x 5.2 fold at 48 hours). Neither mRNA for glutathione peroxydase, gamma-glutamylcysteine synthase and glutathione reductase, nor thioredoxine and thioredoxine reductase, were significantly modified. Additional experiments in which RAW 264.7 cells were stimulated with LPS and/or IFN-gamma in the presence of relatively specific inhibitors of both Mn and Cu/Zn SOD, aminotriazol (ATZ) catalase inhibitor and buthionine sulfoximine (BSO) glutathione inhibitor, showed that inhibiting LPS-induced up-regulation of intracellular redox buffering systems also prevented acquisition of the resistant phenotype.
  
  
  CONCLUSIONS
  Our data suggest a direct causal relationship between survival of a fraction of macrophages and a up-regulation of key sets of auto-protective intracellular redox buffering systems, occurring simultaneously with modulation of expression of apoptotic molecules of the Bcl2-Bcl-XL/Bax-Bad family.
authors.affiliation: Université René Descartes, Paris V Inserm U477, Hôpital Cochin, 75014 Paris, France. pjferret@gmx.net
authors.forename: Pierre-Jacques
authors.initials: PJ
authors.lastname: Ferret
authors.forename: Emmanuelle
authors.initials: E
authors.lastname: Soum
authors.forename: Olivier
authors.initials: O
authors.lastname: Negre
authors.forename: Didier
authors.initials: D
authors.lastname: Fradelizi
mesh_terms: D000818:Animals; D002021:Buffers; D002460:Cell Line; D016898:Interferon-alpha / Q000378:metabolism*; D008070:Lipopolysaccharides / Q000494:pharmacology; D008262:Macrophage Activation; D008264:Macrophages / Q000187:drug effects / Q000378:metabolism*; D051379:Mice; D009569:Nitric Oxide / Q000378:metabolism*; D010084:Oxidation-Reduction; D017382:Reactive Oxygen Species / Q000378:metabolism
chemical_list: D002021:Buffers; D016898:Interferon-alpha; D008070:Lipopolysaccharides; D017382:Reactive Oxygen Species; D009569:Nitric Oxide