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Papers In Press, published online ahead of print April 20, 2000
Neuroscience, DIBIT-H San Raffaele Scientific Institute, Milan 20132
Corresponding Author: clementi.emilio{at}hsr.it
Tumour Necrosis Factor-a (TNFa)-induced maturation of dendritic cells (DC), with downregulation of their endocytic ability, has been reported to be mediated by the accumulation of the lipid messenger ceramide. We have now studied the effects and mechanisms of action of nitric oxide (NO) on endocytosis, investigated with fluorescein-iso-thiocyanate-labelled dextran, using human, monocyte-derived DC, both immature and after treatment with TNFa. Exposure of DC to NO, released by either bystander phagocytes or NO donors, reversed the inhibition of endocytosis induced by TNFa. The intracellular accumulation of ceramide induced by TNFa was also inhibited by NO. In addition, NO was found to exert an inhibitory effect downstream of the TNFa-triggered ceramide accumulation, since NO donors reversed the inhibition of endocytosis induced by the cell-permeant C2-ceramide. These effects of NO were mimicked by the membrane-permeant cyclic GMP analogue, 8-Br cyclic GMP, and prevented by inhibition of the soluble guanylyl cyclase. At variance with rodents, the inducible isoform of the NO synthase was expressed neither in immature human DC, nor after cell treatment with TNFa, interferon-g and lipopolysaccharide, suggesting that regulation of these cells depends on exogenous NO. NO, working through cyclic GMP, might therefore prolong the ability of human DC to internalise antigens at the site of inflammation, and thus modulate the initial steps leading to antigen-specific immune responses.
J. Biol. Chem, 10.1074/jbc.M000511200
Submitted on January 24, 2000
Revised on March 15, 2000
Accepted on April 20, 2000
Nitric oxide inhibits the tumour necrosis factor a-regulated endocytosis of human dendritic cells in a cyclic GMP-dependent way
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