title: Neurotoxic factors released by stimulated human monocytes and THP-1 cells.
abstract: Activated monocytes/macrophages are known to release toxic materials. Identification of these materials is important for developing more effective treatments for inflammatory disorders where self attack occurs. We stimulated human monocytes and THP-1 cells with LPS/IFNγ and measured the toxic effects of their conditioned media against differentiated human NT-2 cells. Their cytotoxicity, as measured by LDH release, was reduced by half when their conditioned media was passed through a 3kDa cutoff filter, indicating an equal division between high and low molecular weight materials. When the high molecular weight components tumor necrosis factor-α (TNFα), interleukin-1β (IL-1β), and IL-6 were removed from the conditioned medium by specific antibodies, the toxicity was reduced by 37-38%. When prostaglandin production was blocked by treatment with the COX inhibitors acetylsalicylic acid and ibuprofen, toxicity was reduced by 15-16%. When oxygen free radical production was blocked by the NADPH inhibitor diphenylene iodonium (DPI) the toxicity was reduced by 17-18%. Treatment with the nitric oxide scavenger carboxy-phenyl-tetramethylimidazolineoxyl-oxide, or the NOS inhibitor N(G)-monomethylene-l-arginine, attenuated the toxicity by about 20%. Removal of released glutamate by glutamate decarboxylase also attenuated the toxicity by 12-13%. In combination, these treatments reduced the toxicity by approximately 50% accounting for the low molecular weight component toxicity. About 10% of the overall toxicity, which was associated with the high molecular weight component, was not identified. Optimal antiinflammatory therapy may require combined suppression of these identified toxin-generating pathways as well as relatively minor pathways yet to be identified.
authors.affiliation: Kinsmen Laboratory of Neurological Research, Vancouver, BC, Canada.
authors.forename: Moonhee
authors.initials: M
authors.lastname: Lee
authors.forename: Kyoungho
authors.initials: K
authors.lastname: Suk
authors.forename: Yunhee
authors.initials: Y
authors.lastname: Kang
authors.forename: Edith
authors.initials: E
authors.lastname: McGeer
authors.forename: Patrick L
authors.initials: PL
authors.lastname: McGeer
mesh_terms: D000704:Analysis of Variance; D000906:Antibodies / Q000494:pharmacology; D002461:Cell Line, Transformed; D045744:Cell Line, Tumor; D003497:Cyclic N-Oxides / Q000494:pharmacology; D051545:Cyclooxygenase 1 / Q000235:genetics / Q000378:metabolism*; D051546:Cyclooxygenase 2 / Q000235:genetics / Q000378:metabolism*; D016207:Cytokines / Q000276:immunology / Q000378:metabolism / Q000494:pharmacology; D015232:Dinoprostone / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D005968:Glutamate Decarboxylase / Q000378:metabolism; D018698:Glutamic Acid / Q000378:metabolism; D006801:Humans; D007093:Imidazoles / Q000494:pharmacology; D007371:Interferon-gamma / Q000633:toxicity; D007770:L-Lactate Dehydrogenase / Q000378:metabolism*; D007948:Leukemia, Monocytic, Acute / Q000188:drug therapy / Q000378:metabolism*; D008070:Lipopolysaccharides / Q000633:toxicity; D008970:Molecular Weight; D009000:Monocytes / Q000187:drug effects / Q000378:metabolism*; D019255:NADPH Oxidases / Q000378:metabolism; D009498:Neurotoxins / Q000494:pharmacology; D052248:Nitric Oxide Synthase Type I / Q000235:genetics / Q000378:metabolism; D009573:Nitrites / Q000378:metabolism; D011758:Pyrroles; D012333:RNA, Messenger / Q000378:metabolism; D018243:Teratocarcinoma / Q000473:pathology
chemical_list: C556060:6,7-dihydro-5H-pyrrolo(1,2-a)imidazole; D000906:Antibodies; D003497:Cyclic N-Oxides; D016207:Cytokines; D007093:Imidazoles; D008070:Lipopolysaccharides; D009498:Neurotoxins; D009573:Nitrites; D011758:Pyrroles; D012333:RNA, Messenger; C079391:2-phenyl-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide; D018698:Glutamic Acid; D007371:Interferon-gamma; D007770:L-Lactate Dehydrogenase; D052248:Nitric Oxide Synthase Type I; D051545:Cyclooxygenase 1; D051546:Cyclooxygenase 2; D019255:NADPH Oxidases; D005968:Glutamate Decarboxylase; C401141:glutamate decarboxylase 2; D015232:Dinoprostone