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  • Title: Failure to induce nitric oxide production by human monocyte-derived macrophages. Manipulation of biochemical pathways.
    Author: Arias M, Zabaleta J, Rodríguez JI, Rojas M, París SC, García LF.
    Journal: Allergol Immunopathol (Madr); 1997; 25(6):280-8. PubMed ID: 9469204.
    Abstract:
    Production of nitrix oxide (NO-) by human macrophages is controversial. In the present study, the ability of human monocyte-derived macrophages (M phi) to produce NO- in response to M phi modulators was tested. M phi cultured for up to nine days and stimulated for 48 with different concentrations of LPS and/or IFN-gamma failed to produce significant amounts of NO2- compared to unstimulated cultures. Inhibition of the cyclo-oxygenase pathway with indomethacin did not increase NO2- production by LPS stimulated M phi. Since human M phi lack biopterin, needed for NO- synthesis by murine M phi, human M phi stimulated with LPS plus IFN-gamma were additionally cultured in the presence of neopterin or biopterin. These treatments did not induce NO2- production. Furthermore, simultaneous treatment with indomethacin and neopterin or biopterin also failed to induce NO2- production. However, human M phi, stimulated with IFN-gamma and LPs, produced TNF-alpha suggesting that the lack of increment in NO2- production was not due to an absence of response of M phi to the stimuli used. As an indirect approach to explore the NO- production, human M phi were infected with virulent Mycobacterium tuberculosis H37Rv and simultaneously treated with the competitive inhibitor NGmonomethyl-L-arginine (NGMMA). Mycobacterial intracellular replication was measured by 3H-uracil incorporation. NGMMA did not have any effect on mycobacterial replication. These results further suggest that human M phi do not produce NO- at least by the inducible pathway.
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