title: Nitric oxide production in mouse and rat macrophages: a rapid and efficient assay for screening of drugs immunostimulatory effects in human cells.
abstract: Activation of inducible nitric oxide (NO) synthase (iNOS) and resulting high-output NO release is known to depend on the action of cytokines. We investigated in vitro production of NO by resident peritoneal macrophages from mice and rats, and secretion of cytokines by these cells as well as by human peripheral blood mononuclear cells (PBMC). The cells were cultured in the presence of a selected group of acyclic nucleoside phosphonates that have previously been shown to possess immunobiological potential. Several of the compounds enhanced production of NO in animal macrophages. This activity was associated with stimulatory effects on secretion of cytokines such as TNF-alpha in all mouse and rat macrophages and human PBMC, and IL-10 in mouse and human cells. Statistically highly significant correlation between the range of NO biosynthesis in rodent cells and extent of cytokine stimulation in human PBMC has been observed. It is suggested that the NO assay may be regarded as an efficient, economical and relatively reliable tool in primary screening for intrinsic immunostimulatory activity of compounds in human cell system, at least from the point of view of cytokine secretion.
authors.affiliation: Institute of Experimental Medicine, Department of Pharmacology, Academy of Sciences, v.v.i., Vídenská 1083, 142 20 Prague 4, Czech Republic.
authors.forename: Eva
authors.initials: E
authors.lastname: Kmonícková
authors.forename: Petra
authors.initials: P
authors.lastname: Melkusová
authors.forename: Hassan
authors.initials: H
authors.lastname: Farghali
authors.forename: Antonín
authors.initials: A
authors.lastname: Holý
authors.forename: Zdenek
authors.initials: Z
authors.lastname: Zídek
mesh_terms: D000225:Adenine / Q000494:pharmacology; D000276:Adjuvants, Immunologic / Q000494:pharmacology*; D000818:Animals; D002478:Cells, Cultured; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D005260:Female; D006801:Humans; D007371:Interferon-gamma / Q000378:metabolism / Q000494:pharmacology; D016753:Interleukin-10 / Q000378:metabolism; D007963:Leukocytes, Mononuclear / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D008070:Lipopolysaccharides / Q000494:pharmacology; D017737:Macrophages, Peritoneal / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D009569:Nitric Oxide / Q000378:metabolism*; D009573:Nitrites / Q000378:metabolism; D063065:Organophosphonates / Q000494:pharmacology; D051381:Rats; D011917:Rats, Inbred Lew; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism
chemical_list: D000276:Adjuvants, Immunologic; D008070:Lipopolysaccharides; C120401:N(6)-cyclopropyl-9-(2-phosphonylmethoxyethyl)-2,6-diaminopurine; D009573:Nitrites; D063065:Organophosphonates; D014409:Tumor Necrosis Factor-alpha; C482199:lipopolysaccharide, E coli O55-B5; D016753:Interleukin-10; D009569:Nitric Oxide; D007371:Interferon-gamma; D000225:Adenine