Quantitative nitric oxide production by rat, bovine and porcine macrophages.
The aim of this work was to compare in vitro nitric oxide (NO) production by rat, bovine and porcine macrophages. NO production was induced by lipopolysaccharide (LPS) or by phorbol 12-myristate 13-acetate (PMA) with ionomycin or recombinant interferon gamma (rIFN-gamma) and was assessed by Griess reaction. NO synthase type II (NOS II) expression was quantified by immunocytochemistry, Western blot and real-time polymerase chain reaction (RT-PCR). There were differences in NO production by pulmonary alveolar macrophages (PAM) in all species tested. The largest amounts of NO were produced by rat PAM. Less NO was produced by bovine PAM. Moreover, PAM in rats and cows differed in their abilities to respond to various stimulators. Neither porcine PAM nor Kupffer cells produced NO. Stimulation of porcine PAM with alternative concentrations of LPS did not lead to inducing NO production. Stimulation of porcine PAM with rIFN-gamma together with LPS led to a significant increase in the expression of NOS II mRNA, albeit without detectable NO production or NOS II expression on the protein level.
Zelnickova
Petra
P
Department of Immunology, Veterinary Research Institute, Hudcova 70, 621 00 Brno, Czech Republic.
Matiasovic
Jan
J
Pavlova
Barbora
B
Kudlackova
Hana
H
Kovaru
Frantisek
F
Faldyna
Martin
M
D000818:Animals; D001681:Biological Assay; D015153:Blotting, Western; D002417:Cattle; D002478:Cells, Cultured; D005029:Ethylenediamines; D015971:Gene Expression Regulation, Enzymologic; D007150:Immunohistochemistry; D007371:Interferon-gamma / Q000633:toxicity; D015759:Ionomycin / Q000633:toxicity; D008070:Lipopolysaccharides / Q000633:toxicity; D016676:Macrophages, Alveolar / Q000378:metabolism / Q000473:pathology; D009569:Nitric Oxide / Q000032:analysis / Q000378:metabolism; D052247:Nitric Oxide Synthase Type II / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D011994:Recombinant Proteins; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013424:Sulfanilamides; D013552:Swine; D013755:Tetradecanoylphorbol Acetate / Q000633:toxicity
D005029:Ethylenediamines; C095000:Griess reagent; D008070:Lipopolysaccharides; D012333:RNA, Messenger; D011994:Recombinant Proteins; D013424:Sulfanilamides; D009569:Nitric Oxide; D015759:Ionomycin; D007371:Interferon-gamma; D052247:Nitric Oxide Synthase Type II; D013755:Tetradecanoylphorbol Acetate