Nitric Oxide Inhibits the Tumor Necrosis Factor α-regulated Endocytosis of Human Dendritic Cells in a Cyclic GMP-dependent Way

Paolucci, Clara; Rovere, Patrizia; Nadaï, C. De; Manfredi, Angelo A.; Clementi, Emilio

doi:10.1074/jbc.m000511200

Cited by 48 publications

(64 citation statements)

References 34 publications

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“…The mtDNA levels were normalized to the protein content, which was measured by using the bicinchoninic acid protein-assay procedure (Perbio, Bezons, France). Analysis of the cellular content of mitochondrial proteins was carried out by flow cytometry, as described (26). Briefly, cell suspensions were permeabilized for 20 min at room temperature in the presence of 1% BSA and 0.1% saponin.…”

Section: Methodsmentioning

confidence: 99%

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Mitochondrial biogenesis by NO yields functionally active mitochondria in mammals

Nisoli

Falcone

Tonello

et al. 2004

Proc. Natl. Acad. Sci. U.S.A.

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460

397

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We recently found that long-term exposure to nitric oxide (NO) triggers mitochondrial biogenesis in mammalian cells and tissues by activation of guanylate cyclase and generation of cGMP. Here, we report that the NO/cGMP-dependent mitochondrial biogenesis is associated with enhanced coupled respiration and content of ATP in U937, L6, and PC12 cells. The observed increase in ATP content depended entirely on oxidative phosphorylation, because ATP formation by glycolysis was unchanged. Brain, kidney, liver, heart, and gastrocnemius muscle from endothelial NO synthase null mutant mice displayed markedly reduced mitochondrial content associated with significantly lower oxygen consumption and ATP content. In these tissues, ultrastructural analyses revealed significantly smaller mitochondria. Furthermore, a significant reduction in the number of mitochondria was observed in the subsarcolemmal region of the gastrocnemius muscle. We conclude that NO/cGMP stimulates mitochondrial biogenesis, both in vitro and in vivo, and that this stimulation is associated with increased mitochondrial function, resulting in enhanced formation of ATP

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Section: Methodsmentioning

confidence: 99%

“…Samples (1 ϫ 10 6 cells) were then incubated with the appropriate primary Abs for 30 min at 4°C. Expression of these molecules was analyzed by flow cytometry after staining with appropriate fluorescein isothyocyanate-labeled Abs, by using a fluorescenceactivated cell sorter (FACStar Plus, Beckton Dickinson) (26).…”

Section: Methodsmentioning

confidence: 99%

Mitochondrial biogenesis by NO yields functionally active mitochondria in mammals

Nisoli

Falcone

Tonello

et al. 2004

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

460

397

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“…Receptor-mediated endocytosis was also shown for IFN-γ, but in contrast to the recycling of the IL-1β receptor, a lysosomal ligandreceptor degradation occurred after internalisation [73]. Endocytotic internalisation of cytokine receptors is also modulated by NO, which had a negative effect upon TNF receptor endocytosis in dendritic cells [74,75]. To our knowledge, the internalisation of cytokine receptors has not been investigated systematically in pancreatic beta cells by functional studies.…”

Section: Discussionmentioning

confidence: 99%

Cytokines activate genes of the endocytotic pathway in insulin-producing RINm5F cells

et al. 2004

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Aims/hypothesis. Cytokines are important humoral mediators of beta cell destruction in autoimmune diabetes. The aim of this study was to identify novel cytokine-induced genes in insulin-producing RINm5F cells, which may contribute to beta cell death or survival. Methods. A global gene expression profile in cytokine-exposed insulin-producing RINm5F cells was achieved by automated restriction fragment differential display PCR. The expression of selected candidate genes was confirmed by real-time RT-PCR analysis. Results. Exposure of RINm5F cells to IL-1β or to a cytokine mixture (IL-1β, TNF-α, IFN-γ) for 6 h resulted in the differential expression of a functional gene cluster. Apart from the well-known up-regulation of the cytokine-responsive genes iNOS, NF-κB, MnSOD and Hsp70, several genes that belong to the functional cluster of the endocytotic pathway were identified. These endocytotic genes comprised: clathrin, megalin, synaptotagmin and calcineurin, which were up-regulated by IL-1β or the cytokine mixture.In contrast, the expression of the calcineurin inhibitor CAIN and of the GDP/GTP exchange protein Rab3 was down-regulated by cytokines. Other up-regulated cytokine-responsive genes were: agrin, murine adherent macrophage protein mRNA (MAMA) and transport-associated protein (TAP1/MTP), whereas the plasma membrane calcium ATPase (PMCA) 2 and PMCA 3 genes were down-regulated by cytokines. Conclusions/interpretation. Our results indicate that genes of the endocytotic pathway are regulated by pro-inflammatory cytokines. This might affect the density of cytokine receptors at the beta cell surface and concomitantly the sensitivity of the cells to cytokine toxicity. A better understanding of the functional cross-talk between endocytotic and cytokine signalling pathways could further the development of novel strategies to protect pancreatic beta cells against toxic effects of pro-inflammatory cytokines.

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“…For flow cytometry, Abs against human CD2 (6F10.3), CD14 (RM052), CD19 (J4.119), CD80 (MAB104), CD83 (HB15A) (all Beckmann Coulter, Krefeld, Germany), CD86 (BU63, Serotec, Oxford, U.K.), HLA-DR (YD1/ 63.4.10, Serotec), nNOS (nNOS-AB, A-11, Santa Cruz Biotechnology, Santa Cruz, CA), CD40 (5c3), CD208-APC (110-1112) (all Becton Dickinson), CCR7-APC (150503; R&D Systems, Wiesbaden, Germany), mouse (MOPC-31-C, MOPC-173, [27][28][29][30][31][32][33][34][35], and rat (R35-95, MOPC-21) subclassspecific isotypes (all Beckmann Coulter) were used. Conjugated secondary reagents: FITC-conjugated goat-anti-rat-IgG (Biozol, Eching, Germany), PE-conjugated donkey anti-mouse-IgG (Jackson ImmunoResearch Laboratories, Suffolk, U.K.), for myeloid DC: anti-CD1c-PE (M241, Ancell, Cologne, Germany).…”

Section: Absmentioning

confidence: 99%

“…presence of NO-GC and NO-stimulated cGMP generation has been reported (35)(36)(37). To determine whether the effect of NO on DC maturation is mediated by cGMP, DCs were treated with the NO-GC-specific inhibitor ODQ during generation of iDCs (days 1-6 of culture, Fig.…”

Section: The Effect Of No On Human DC Maturation Is Cgmp-dependentmentioning

confidence: 99%

Neuronal Nitric Oxide Synthase Modulates Maturation of Human Dendritic Cells

Adler¹,

Simon

Graulich³

et al. 2010

The Journal of Immunology

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Dendritic cells (DCs) are the most potent APCs of the immune system. Understanding the intercellular and intracellular signaling processes that lead to DC maturation is critical for determining how these cells initiate T cell-mediated immune processes. NO synthesized by the inducible NO synthase (iNOS) is important for the function of murine DCs. In our study, we investigated the regulation of the arginine/NO-system in human monocyte-derived DCs. Maturation of DCs induced by inflammatory cytokines (IL-1β, TNF, IL-6, and PGE2) resulted in a pronounced expression of neuronal NOS (nNOS) but only minimal levels of iNOS and endothelial NOS were detected in human mature DCs. In addition, reporter cell assays revealed the production of NO by mature DCs. Specific inhibitors of NOS (N-nitro-l-arginine methyl ester) or of the NO target guanylyl cyclase (H-(1,2,4)-oxadiazolo [4,3-a] quinoxalin-1-one) prevented DC maturation (shown by decreased expression of MHC class II, costimulatory and CD83 molecules and reduced IL-12 production) and preserved an immature phenotype, indicating an autocrine effect of nNOS-derived NO on human DC maturation. Notably, inhibitor-treated DCs were incapable of inducing efficient T cell responses after primary culture and generated an anergic T cell phenotype. In conclusion, our results suggest that, in the human system, nNOS-, but not iNOS-derived NO, plays an important regulatory role for the maturation of DCs and, thus, the induction of pronounced T cell responses.

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Nitric Oxide Inhibits the Tumor Necrosis Factor α-regulated Endocytosis of Human Dendritic Cells in a Cyclic GMP-dependent Way

Cited by 48 publications

References 34 publications

Mitochondrial biogenesis by NO yields functionally active mitochondria in mammals

Mitochondrial biogenesis by NO yields functionally active mitochondria in mammals

Cytokines activate genes of the endocytotic pathway in insulin-producing RINm5F cells

Neuronal Nitric Oxide Synthase Modulates Maturation of Human Dendritic Cells

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