Oxygen radicals and nitric oxide production by turkey respiratory macrophages.
The influence of different induction protocols on the recovery of elicited turkey respiratory macrophages (RM), and on their oxygenation activity and nitric oxide (NO) production was examined. RM were induced in three week old specific pathogen free turkeys with Sephadex G-50, Thioglycollate broth, and an emulsion of incomplete Freund's adjuvant (IFA), supplemented either with Mycoplasma hyorhinis grown in Modified Channock broth (IFA-M. hyorhinis) or with Modified Channock broth (IFA-Broth). The RM were recovered by lavage of the lungs and air sacs and were purified by centrifugation through a Percoll suspension. Their oxygenation activity was evaluated in luminol-enhanced chemiluminescence assays, following stimulation with Zymosan A. The NO production was evaluated by incubating the RM with lipopolysaccharide (LPS) from Salmonella enteritidis for 24 or 48 hours. The number of recovered RM was slightly, but not significantly lower for Sephadex G-50 and IFA-Broth than for Thioglycollate broth and IFA-M. hyorhinis. RM elicited with Sephadex G-50 and IFA-Broth showed a significantly higher oxidative burst response to Zymosan A, compared to the Thioglycollate and IFA-M. hyorhinis elicited RM. Although all elicited RM showed a high NO production upon stimulation with LPS, no significant differences were seen in the NO production of the RM obtained following the different induction treatments. Our results point out that care should be taken when applying elicited RM for in vitro assays, as distinct levels of oxygenation activity were obtained using different induction protocols.
Van Nerom
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AE
Department of Pathology, Bacteriology and Avian Diseases, Faculty of Veterinary Medicine, University of Ghent, Belgium. anne.vannerom@rug.ac.be
Ducatelle
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R
Haesebrouck
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F
D000818:Animals; D002469:Cell Separation; D003911:Dextrans; D008070:Lipopolysaccharides / Q000494:pharmacology; D008163:Luminescent Measurements; D008165:Luminol; D016676:Macrophages, Alveolar / Q000378:metabolism*; D009569:Nitric Oxide / Q000096:biosynthesis*; D010100:Oxygen / Q000378:metabolism*; D016897:Respiratory Burst / Q000187:drug effects; D012477:Salmonella enteritidis; D014422:Turkeys; D015054:Zymosan / Q000494:pharmacology
D003911:Dextrans; D008070:Lipopolysaccharides; D009569:Nitric Oxide; D008165:Luminol; D015054:Zymosan; C025614:sephadex; D010100:Oxygen