Induction of nitric oxide synthase in bovine mononuclear phagocytes is differentiation stage-dependent

Bovine monocytes and monocyte-derived macrophages (MDM) activated by various means were assessed for induction of inducible nitric oxide synthase (iNOS), using the Griess assay, Northern blotting and reverse transcription/polymerase chain reaction (RT-PCR). Interferon-gamma (IFN-gamma) induced little, if any, iNOS expression and NO production in MDM, although these cells responded to IFN-gamma in other regards. In contrasts, MDM produced copious amounts of NO when stimulated with LPS or Salmonella dublin, and this was paralleled by high steady state levels of iNOS mRNA. Heat-killed Listeria monocytogenes induced more iNOS mRNA and nitrite than IFN-gamma, but much less than L. mono-cytogenes and IFN-gamma combined. Monocytes differed from M phi with respect to iNOS induction and nitrite production in several regards: (i) LPS and S. dublin induced only low levels of iNOS mRNA and nitrite in monocytes, although cells responded to these stimuli in various other ways: (ii) IFN-gamma alone induced in monocytes iNOS mRNA generation and NO formation, although to a low and variable degree; (iii) upon maximal stimulation (e.g. by L. monocytogenes and IFN-gamma combined), monocytes produced much less nitrite than MDM, and mRNA levels were lower. Regulation of macrophage iNOS varies considerably between species. We provide the first evidence in any species that the steady state levels of iNOS mRNA and NO generation in monocytes and macrophages activated by various means depend on the stage of mononuclear phagocyte differentiation.