14 results on '"Meijler, Michael M."'
Search Results
2. Exogenous ceramide-1-phosphate (C1P) and phospho-ceramide analogue-1 (PCERA-1) regulate key macrophage activities via distinct receptors.
- Author
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Katz S, Ernst O, Avni D, Athamna M, Philosoph A, Arana L, Ouro A, Hoeferlin LA, Meijler MM, Chalfant CE, Gómez-Muñoz A, and Zor T
- Subjects
- ADAM Proteins metabolism, ADAM17 Protein, Animals, Cell Line, Cell Movement drug effects, Cyclic AMP metabolism, Cyclic AMP Response Element-Binding Protein metabolism, Drug Synergism, Gene Expression Regulation drug effects, Inflammation immunology, Interleukin-10 genetics, Interleukin-10 metabolism, Lipopolysaccharides immunology, Macrophages immunology, Mice, Receptors, G-Protein-Coupled metabolism, Signal Transduction drug effects, Tumor Necrosis Factor-alpha genetics, Tumor Necrosis Factor-alpha metabolism, Ceramides pharmacology, Inflammation drug therapy, Macrophages drug effects
- Abstract
Inflammation is an ensemble of tightly regulated steps, in which macrophages play an essential role. Previous reports showed that the natural sphingolipid ceramide 1-phosphate (C1P) stimulates macrophages migration, while the synthetic C1P mimic, phospho-ceramide analogue-1 (PCERA-1), suppresses production of the key pro-inflammatory cytokine TNFα and amplifies production of the key anti-inflammatory cytokine IL-10 in LPS-stimulated macrophages, via one or more unidentified G-protein coupled receptors. We show that C1P stimulated RAW264.7 macrophages migration via the NFκB pathway and MCP-1 induction, while PCERA-1 neither mimicked nor antagonized these activities. Conversely, PCERA-1 synergistically elevated LPS-dependent IL-10 expression in RAW264.7 macrophages via the cAMP-PKA-CREB signaling pathway, while C1P neither mimicked nor antagonized these activities. Interestingly, both compounds have the capacity to additively inhibit TNFα secretion; PCERA-1, but not C1P, suppressed LPS-induced TNFα expression in macrophages in a CREB-dependent manner, while C1P, but not PCERA-1, directly inhibited recombinant TNFα converting enzyme (TACE). Finally, PCERA-1 failed to interfere with binding of C1P to either the cell surface receptor or to TACE. These results thus indicate that the natural sphingolipid C1P and its synthetic analog PCERA-1 bind and activate distinct receptors expressed in RAW264.7 macrophages. Identification of these receptors will be instrumental for elucidation of novel activities of extra-cellular sphingolipids, and may pave the way for the design of new sphingolipid mimics for the treatment of inflammatory diseases, and pathologies which depend on cell migration, as in metastatic tumors., (Copyright © 2015 Elsevier B.V. All rights reserved.)
- Published
- 2016
- Full Text
- View/download PDF
3. A ceramide analog inhibits cPLA(2) activity and consequent PGE(2) formation in LPS-stimulated macrophages.
- Author
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Goldsmith M, Daka A, Lamour NF, Mashiach R, Glucksam Y, Meijler MM, Chalfant CE, and Zor T
- Subjects
- Animals, Cell Line, Dinoprostone biosynthesis, Group IV Phospholipases A2 immunology, Group IV Phospholipases A2 pharmacology, Lipopolysaccharides, Macrophages enzymology, Male, Mice, Mice, Inbred BALB C, Ceramides pharmacology, Dinoprostone immunology, Group IV Phospholipases A2 antagonists & inhibitors, Macrophages immunology
- Abstract
Prostaglandin E(2) (PGE(2)) is an important mediator of the inflammatory response. Phospho-ceramide analogue-1 (PCERA-1), a synthetic phospholipid-like molecule, was previously reported to modulate pro- and anti-inflammatory cytokine production. We show here that PCERA-1 inhibited LPS-stimulated PGE(2) production in RAW264.7 macrophages, without affecting COX-2 expression. Furthermore, PCERA-1 efficiently suppressed arachidonic acid (AA) release in response to LPS. The dephosphorylated derivative of PCERA-1, ceramide analogue-1 (CERA-1), mimicked the inhibitory effect of PCERA-1 on AA release and PGE(2) production in macrophages. Inhibition of PGE(2) production by CERA-1 was completely rescued by addition of exogenous AA. Importantly, PCERA-1 and ceramide-1-phosphate (C1P) stimulated the enzymatic activity of cPLA(2)α in an in vitro assay, whereas CERA-1 and ceramide inhibited both basal and C1P-stimulated cPLA(2)α activity. Collectively, these results indicate that CERA-1 suppresses AA release and subsequent PGE(2) production in LPS-stimulated macrophages by direct interaction with cPLA(2), and suggest that ceramide may similarly counteract C1P effect on cPLA(2) activity in cells. The suppression of PGE(2) production is suggested to contribute to the anti-inflammatory action of PCERA-1., (Copyright © 2010 Elsevier B.V. All rights reserved.)
- Published
- 2011
- Full Text
- View/download PDF
4. The ceramide-1-phosphate analogue PCERA-1 modulates tumour necrosis factor-alpha and interleukin-10 production in macrophages via the cAMP-PKA-CREB pathway in a GTP-dependent manner.
- Author
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Avni D, Philosoph A, Meijler MM, and Zor T
- Subjects
- Adenylyl Cyclases metabolism, Animals, Anti-Inflammatory Agents, Non-Steroidal pharmacology, Bucladesine pharmacology, Calcium Signaling drug effects, Cell Line, Cell Membrane drug effects, Cell Membrane metabolism, Cyclic AMP analogs & derivatives, Cyclic AMP metabolism, Cyclic AMP Response Element-Binding Protein metabolism, Cyclic AMP-Dependent Protein Kinases antagonists & inhibitors, Cyclic AMP-Dependent Protein Kinases metabolism, Dinoprostone pharmacology, Enzyme Inhibitors pharmacology, Genes, Reporter genetics, Imidazoles pharmacology, Indoles pharmacology, Isoquinolines pharmacology, Lipopolysaccharides pharmacology, Macrophages drug effects, Mice, Phosphorylation drug effects, Protein Kinase Inhibitors pharmacology, Pyridines pharmacology, Response Elements genetics, Ribosomal Protein S6 Kinases, 90-kDa antagonists & inhibitors, Ribosomal Protein S6 Kinases, 90-kDa metabolism, Rolipram pharmacology, Signal Transduction physiology, Sulfonamides pharmacology, Transfection, p38 Mitogen-Activated Protein Kinases antagonists & inhibitors, p38 Mitogen-Activated Protein Kinases metabolism, Ceramides pharmacology, Guanosine Triphosphate metabolism, Interleukin-10 metabolism, Macrophages metabolism, Signal Transduction drug effects, Tumor Necrosis Factor-alpha metabolism
- Abstract
The synthetic phospho-ceramide analogue-1 (PCERA-1) down-regulates production of the pro-inflammatory cytokine tumour necrosis factor-alpha (TNF-alpha) and up-regulates production of the anti-inflammatory cytokine interleukin-10 (IL-10) in lipopolysaccharide (LPS) -stimulated macrophages. We have previously reported that PCERA-1 increases cyclic adenosine monophosphate (cAMP) levels. The objective of this study was to delineate the signalling pathway leading from PCERA-1 via cAMP to modulation of TNF-alpha and IL-10 production. We show here that PCERA-1 elevates intra-cellular cAMP level in a guanosine triphosphate-dependent manner in RAW264.7 macrophages. The cell-permeable dibutyryl cAMP was able to mimic the effects of PCERA-1 on cytokine production, whereas 8-chloro-phenylthio-methyladenosine-cAMP, which specifically activates the exchange protein directly activated by cAMP (EPAC) but not protein kinase A (PKA), failed to mimic PCERA-1 activities. Consistently, the PKA inhibitor H89 efficiently blocked PCERA-1-driven cytokine modulation as well as PCERA-1-stimulated phosphorylation of cAMP response element binding protein (CREB) on Ser-133. Finally, PCERA-1 activated cAMP-responsive transcription of a luciferase reporter, in synergism with the phosphodiesterase (PDE)-4 inhibitor rolipram. Our results suggest that PCERA-1 activates a G(s) protein-coupled receptor, leading to elevation of cAMP, which acts via the PKA-CREB pathway to promote TNF-alpha suppression and IL-10 induction in LPS-stimulated macrophages. Identification of the PCERA-1 receptor is expected to set up a new target for development of novel anti-inflammatory drugs.
- Published
- 2010
- Full Text
- View/download PDF
5. Distinct receptor-mediated activities in macrophages for natural ceramide-1-phosphate (C1P) and for phospho-ceramide analogue-1 (PCERA-1).
- Author
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Levi M, Meijler MM, Gómez-Muñoz A, and Zor T
- Subjects
- Animals, Cell Movement immunology, Ceramides immunology, Humans, Inflammation immunology, Inflammation metabolism, Macrophages immunology, Signal Transduction immunology, Ceramides metabolism, Macrophages metabolism
- Abstract
Ceramide-1-phosphate (C1P) is known as a second messenger regulating a multitude of processes including cell growth, apoptosis and inflammation. Exciting recent findings now suggest that C1P can stimulate macrophages migration in an extra-cellular manner via a G protein-coupled receptor (GPCR). Interestingly, a synthetic C1P analog, named phospho-ceramide analogue-1 (PCERA-1), was recently described as a potent in-vivo anti-inflammatory agent, and was suggested to act on macrophages in an extra-cellular manner via a GPCR. Here we summarize and compare the receptor-mediated as well as receptor-independent activities of natural C1P and its synthetic analog. We also provide experimental data in support of distinct C1P and PCERA-1 receptors., (2009 Elsevier Ireland Ltd. All rights reserved.)
- Published
- 2010
- Full Text
- View/download PDF
6. A ceramide-1-phosphate analogue, PCERA-1, simultaneously suppresses tumour necrosis factor-alpha and induces interleukin-10 production in activated macrophages.
- Author
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Goldsmith M, Avni D, Levy-Rimler G, Mashiach R, Ernst O, Levi M, Webb B, Meijler MM, Gray NS, Rosen H, and Zor T
- Subjects
- Animals, Anti-Inflammatory Agents immunology, Cells, Cultured, Gene Expression Regulation immunology, Inflammation Mediators metabolism, Interleukin-10 genetics, Lipopolysaccharides immunology, Macrophage Activation immunology, Mice, RNA, Messenger genetics, Signal Transduction immunology, Toll-Like Receptors agonists, Toll-Like Receptors immunology, Tumor Necrosis Factor-alpha genetics, Ceramides immunology, Interleukin-10 biosynthesis, Macrophages immunology, Tumor Necrosis Factor-alpha biosynthesis
- Abstract
Tight regulation of the production of the key pro-inflammatory cytokine tumour necrosis factor-alpha (TNF-alpha) is essential for the prevention of chronic inflammatory diseases. In vivo administration of a synthetic phospholipid, named hereafter phospho-ceramide analogue-1 (PCERA-1), was previously found to suppress lipopolysaccharide (LPS)-induced TNF-alpha blood levels. We therefore investigated the in vitro anti-inflammatory effects of PCERA-1. Here, we show that extracellular PCERA-1 potently suppresses production of the pro-inflammatory cytokine TNF-alpha in RAW264.7 macrophages, and in addition, independently and reciprocally regulates the production of the anti-inflammatory cytokine interleukin-10 (IL-10). Specificity is demonstrated by the inability of the phospholipids ceramide-1-phosphate (C1P), sphingosine-1-phosphate (S1P) and lysophosphatidic acid (LPA) to perform these activities. Similar TNF-alpha suppression and IL-10 induction by PCERA-1 were observed in macrophages when activated by Toll-like receptor 4 (TLR4), TLR2 and TLR7 agonists. Regulation of cytokine production is demonstrated at the mRNA and protein levels. Finally, we show that, while PCERA-1 does not block activation of nuclear factor (NF)-kappaB and mitogen-activated protein kinases by LPS, it elevates the intracellular cAMP level. In conclusion, the anti-inflammatory activity of PCERA-1 seems to be mediated by a cell membrane receptor, upstream of cAMP production, and eventually TNF-alpha suppression and IL-10 induction. Thus, identification of the PCERA-1 receptor may provide new pharmacological means to block inflammation.
- Published
- 2009
- Full Text
- View/download PDF
7. Modulation of TNFalpha, IL-10 and IL-12p40 levels by a ceramide-1-phosphate analog, PCERA-1, in vivo and ex vivo in primary macrophages.
- Author
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Avni D, Goldsmith M, Ernst O, Mashiach R, Tuntland T, Meijler MM, Gray NS, Rosen H, and Zor T
- Subjects
- Animals, Cell Differentiation drug effects, Cell Differentiation immunology, Dinoprostone immunology, Dinoprostone metabolism, Inflammation immunology, Inflammation metabolism, Interferon-gamma pharmacology, Interleukin-10 agonists, Interleukin-12 Subunit p40 antagonists & inhibitors, Lipopolysaccharides pharmacology, Macrophages immunology, Male, Mice, Mice, Inbred BALB C, Monocytes drug effects, Monocytes metabolism, Tumor Necrosis Factor-alpha antagonists & inhibitors, Ceramides pharmacology, Interleukin-10 biosynthesis, Interleukin-12 Subunit p40 biosynthesis, Macrophages drug effects, Monocytes immunology, Tumor Necrosis Factor-alpha biosynthesis
- Abstract
Phospho-ceramide analog-1 (PCERA-1) has been described as a potent in vivo suppressor of the pro-inflammatory cytokine tumor necrosis factor alpha (TNFalpha), and thus as a putative drug for the treatment of inflammatory diseases. However, the in vivo cell target of PCERA-1 has not been identified, and its in vivo effect on secretion of other relevant cytokines has not been reported. We have previously shown that PCERA-1 suppresses lipopolysaccharide (LPS)-induced TNFalpha production in RAW264.7 macrophages in vitro. We therefore hypothesized that PCERA-1 targets TNFalpha production by primary macrophages. In this study we thus investigated the effect of PCERA-1 on LPS-induced release of TNFalpha, interleukin (IL)-10 and IL-12p40, in vivo, and ex vivo. We found that PCERA-1 suppressed production of the pro-inflammatory cytokines, TNFalpha and IL-12p40, and increased production of the anti-inflammatory cytokine, IL-10, in LPS-challenged mice, and in primary peritoneal macrophages as well as bone marrow-derived macrophages (BMDM) stimulated with LPS and interferon (IFN)-gamma. These activities of PCERA-1 were independent of each other. In contrast, PCREA-1 only slightly affected TNFalpha production in the whole blood assay, where LPS-induced cytokines are mainly produced by monocytes. Moreover, isolated blood monocytes were inert to PCERA-1, but acquired responsiveness to PCERA-1 upon macrophage colony stimulating factor (M-CSF)-induced differentiation into macrophages. Pharmacokinetic analysis in mice showed that while the volume of distribution of PCERA-1 is low, the drug was rapidly exchanged between the peritoneum and the systemic circulation. Together, these results suggest that sensitivity to PCERA-1 increases upon differentiation of blood monocytes into tissue macrophages, and imply a mechanistic role for peritoneal macrophages in the in vivo anti-inflammatory activity of PCERA-1. Finally, we show that the mechanism of activity of PCERA-1 and prostaglandin E2 (PGE2) is distinct, and that PCERA-1 signaling is not mediated by EP2, a PGE2 receptor which is also activated by oxidized phospholipids. The independent and reciprocal modulation of production of TNFalpha and IL-12p40, vs. IL-10, suggests that PCERA-1 may be a candidate drug for the treatment of inflammation-linked diseases.
- Published
- 2009
- Full Text
- View/download PDF
8. Modulation of gene expression via disruption of NF-kappaB signaling by a bacterial small molecule.
- Author
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Kravchenko VV, Kaufmann GF, Mathison JC, Scott DA, Katz AZ, Grauer DC, Lehmann M, Meijler MM, Janda KD, and Ulevitch RJ
- Subjects
- 4-Butyrolactone physiology, Adult, Animals, Cyclic AMP Response Element-Binding Protein metabolism, Cystic Fibrosis microbiology, Female, Homoserine physiology, Humans, I-kappa B Kinase metabolism, I-kappa B Proteins metabolism, Immunity, Innate, Interferon-gamma immunology, Lipopolysaccharides immunology, Macrophage Activation, Male, Mice, Mice, Inbred C57BL, Mice, Transgenic, Middle Aged, NF-KappaB Inhibitor alpha, Phosphorylation, Pseudomonas Infections immunology, Pseudomonas Infections microbiology, Pseudomonas aeruginosa immunology, Pseudomonas aeruginosa physiology, Toll-Like Receptors metabolism, Transcription Factor RelA metabolism, 4-Butyrolactone analogs & derivatives, Gene Expression Regulation, Homoserine analogs & derivatives, Macrophages immunology, Macrophages metabolism, NF-kappa B metabolism, Pseudomonas aeruginosa pathogenicity, Signal Transduction
- Abstract
The control of innate immune responses through activation of the nuclear transcription factor NF-kappaB is essential for the elimination of invading microbial pathogens. We showed that the bacterial N-(3-oxo-dodecanoyl) homoserine lactone (C12) selectively impairs the regulation of NF-kappaB functions in activated mammalian cells. The consequence is specific repression of stimulus-mediated induction of NF-kappaB-responsive genes encoding inflammatory cytokines and other immune regulators. These findings uncover a strategy by which C12-producing opportunistic pathogens, such as Pseudomonas aeruginosa, attenuate the innate immune system to establish and maintain local persistent infection in humans, for example, in cystic fibrosis patients.
- Published
- 2008
- Full Text
- View/download PDF
9. N-(3-oxo-acyl)homoserine lactones signal cell activation through a mechanism distinct from the canonical pathogen-associated molecular pattern recognition receptor pathways.
- Author
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Kravchenko VV, Kaufmann GF, Mathison JC, Scott DA, Katz AZ, Wood MR, Brogan AP, Lehmann M, Mee JM, Iwata K, Pan Q, Fearns C, Knaus UG, Meijler MM, Janda KD, and Ulevitch RJ
- Subjects
- 4-Butyrolactone chemistry, 4-Butyrolactone physiology, Amino Acid Motifs, Animals, Bone Marrow Cells cytology, Macrophages metabolism, Mice, Mice, Inbred C57BL, Mice, Knockout, Phosphorylation, Pseudomonas aeruginosa metabolism, RNA, Viral metabolism, Signal Transduction, 4-Butyrolactone analogs & derivatives, Bone Marrow Cells microbiology, Gene Expression Regulation, Macrophages microbiology
- Abstract
Innate immune system receptors function as sensors of infection and trigger the immune responses through ligand-specific signaling pathways. These ligands are pathogen-associated products, such as components of bacterial walls and viral nuclear acids. A common response to such ligands is the activation of mitogen-activated protein kinase p38, whereas double-stranded viral RNA additionally induces the phosphorylation of eukaryotic translation initiation factor 2alpha (eIF2alpha). Here we have shown that p38 and eIF2alpha phosphorylation represent two biochemical markers of the effects induced by N-(3-oxo-acyl)homoserine lactones, the secreted products of a number of Gram-negative bacteria, including the human opportunistic pathogen Pseudomonas aeruginosa. Furthermore, N-(3-oxo-dodecanoyl)homoserine lactone induced distension of mitochondria and the endoplasmic reticulum as well as c-jun gene transcription. These effects occurred in a wide variety of cell types including alveolar macrophages and bronchial epithelial cells, requiring the structural integrity of the lactone ring motif and its natural stereochemistry. These findings suggest that N-(3-oxo-acyl)homoserine lactones might be recognized by receptors of the innate immune system. However, we provide evidence that N-(3-oxo-dodecanoyl)homoserine lactone-mediated signaling does not require the presence of the canonical innate immune system receptors, Toll-like receptors, or two members of the NLR/Nod/Caterpillar family, Nod1 and Nod2. These data offer a new understanding of the effects of N-(3-oxo-dodecanoyl)homoserine lactone on host cells and its role in persistent airway infections caused by P. aeruginosa.
- Published
- 2006
- Full Text
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10. The ceramide-1-phosphate analogue PCERA-1 modulates tumour necrosis factor-α and interleukin-10 production in macrophages via the cAMP–PKA–CREB pathway in a GTP-dependent manner.
- Author
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Avni, Dorit, Philosoph, Amir, Meijler, Michael M., and Zor, Tsaffrir
- Subjects
TUMOR necrosis factors ,CYTOKINES ,INTERLEUKIN-10 ,MACROPHAGES ,ADENOSINE monophosphate ,GUANOSINE triphosphate ,PROTEIN kinases - Abstract
The synthetic phospho-ceramide analogue-1 (PCERA-1) down-regulates production of the pro-inflammatory cytokine tumour necrosis factor-α (TNF-α) and up-regulates production of the anti-inflammatory cytokine interleukin-10 (IL-10) in lipopolysaccharide (LPS) -stimulated macrophages. We have previously reported that PCERA-1 increases cyclic adenosine monophosphate (cAMP) levels. The objective of this study was to delineate the signalling pathway leading from PCERA-1 via cAMP to modulation of TNF-α and IL-10 production. We show here that PCERA-1 elevates intra-cellular cAMP level in a guanosine triphosphate-dependent manner in RAW264.7 macrophages. The cell-permeable dibutyryl cAMP was able to mimic the effects of PCERA-1 on cytokine production, whereas 8-chloro-phenylthio-methyladenosine-cAMP, which specifically activates the exchange protein directly activated by cAMP (EPAC) but not protein kinase A (PKA), failed to mimic PCERA-1 activities. Consistently, the PKA inhibitor H89 efficiently blocked PCERA-1-driven cytokine modulation as well as PCERA-1-stimulated phosphorylation of cAMP response element binding protein (CREB) on Ser-133. Finally, PCERA-1 activated cAMP-responsive transcription of a luciferase reporter, in synergism with the phosphodiesterase (PDE)-4 inhibitor rolipram. Our results suggest that PCERA-1 activates a G
s protein-coupled receptor, leading to elevation of cAMP, which acts via the PKA–CREB pathway to promote TNF-α suppression and IL-10 induction in LPS-stimulated macrophages. Identification of the PCERA-1 receptor is expected to set up a new target for development of novel anti-inflammatory drugs. [ABSTRACT FROM AUTHOR]- Published
- 2010
- Full Text
- View/download PDF
11. A ceramide-1-phosphate analogue, PCERA-1, simultaneously suppresses tumour necrosis factor-α and induces interleukin-10 production in activated macrophages.
- Author
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Goldsmith, Meir, Avni, Dorit, Levy-Rimler, Galit, Mashiach, Roi, Ernst, Orna, Levi, Maya, Webb, Bill, Meijler, Michael M., Gray, Nathanael S., Rosen, Hugh, and Zor, Tsaffrir
- Subjects
CYTOKINES ,INTERLEUKIN-10 ,TUMOR necrosis factors ,PHOSPHOLIPIDS ,INFLAMMATION ,MACROPHAGES - Abstract
Tight regulation of the production of the key pro-inflammatory cytokine tumour necrosis factor-α (TNF-α) is essential for the prevention of chronic inflammatory diseases. In vivo administration of a synthetic phospholipid, named hereafter phospho-ceramide analogue-1 (PCERA-1), was previously found to suppress lipopolysaccharide (LPS)-induced TNF-α blood levels. We therefore investigated the in vitro anti-inflammatory effects of PCERA-1. Here, we show that extracellular PCERA-1 potently suppresses production of the pro-inflammatory cytokine TNF-α in RAW264.7 macrophages, and in addition, independently and reciprocally regulates the production of the anti-inflammatory cytokine interleukin-10 (IL-10). Specificity is demonstrated by the inability of the phospholipids ceramide-1-phosphate (C1P), sphingosine-1-phosphate (S1P) and lysophosphatidic acid (LPA) to perform these activities. Similar TNF-α suppression and IL-10 induction by PCERA-1 were observed in macrophages when activated by Toll-like receptor 4 (TLR4), TLR2 and TLR7 agonists. Regulation of cytokine production is demonstrated at the mRNA and protein levels. Finally, we show that, while PCERA-1 does not block activation of nuclear factor (NF)-κB and mitogen-activated protein kinases by LPS, it elevates the intracellular cAMP level. In conclusion, the anti-inflammatory activity of PCERA-1 seems to be mediated by a cell membrane receptor, upstream of cAMP production, and eventually TNF-α suppression and IL-10 induction. Thus, identification of the PCERA-1 receptor may provide new pharmacological means to block inflammation. [ABSTRACT FROM AUTHOR]
- Published
- 2009
- Full Text
- View/download PDF
12. The Bacterial Quorum-Sensing Signal Molecule N-3-Oxo-Dodecanoyl-L-Homoserine Lactone Reciprocally Modulates Pro- and Anti-Inflammatory Cytokines in Activated Macrophages.
- Author
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Glucksam-Galnoy, Yifat, Sananes, Roy, Silberstein, Nava, Krief, Pnina, Kravchenko, Vladimir V., Meijler, Michael M., and Zor, Tsaffrir
- Subjects
- *
QUORUM sensing , *LACTONES , *CYTOKINES , *MACROPHAGES , *CELLULAR signal transduction , *INTERLEUKIN-10 , *BACTERIA - Abstract
The bacterial molecule N-3-oxo-dodecanoyl-l-homoserine lactone (C12) has critical roles in both interbacterial communication and interkingdom signaling. The ability of C12 to downregulate production of the key proinflammatory cytokine TNF-α in stimulated macrophages was suggested to contribute to the establishment of chronic infections by opportunistic Gram-negative bacteria, such as Pseudomonas aeruginosa. We show that, in contrast to TNF-α suppression, C12 amplifies production of the major anti-inflammatory cytokine IL-10 in LPS-stimulated murine RAW264.7 macrophages, as well as peritoneal macrophages. Furthermore, C12 increased IL-10 mRNA levels and IL-10 promoter reporter activity in LPS-stimulated RAW264.7 macrophages, indicating that C12 modulates IL-10 expression at the transcriptional level. Finally, C12 substantially potentiated LPS-stimulated NF-κB DNA-binding levels and prolonged p38 MAPK phosphorylation in RAW264.7 macrophages, suggesting that increased transcriptional activity of NF-κB and/or p38-activated transcription factors serves to upregulate IL-10 production in macrophages exposed to both LPS and C12. These findings reveal another part of the complex array of host transitions through which opportunistic bacteria downregulate immune responses to flourish and establish a chronic infection. [ABSTRACT FROM AUTHOR]
- Published
- 2013
- Full Text
- View/download PDF
13. A ceramide analog inhibits cPLA2 activity and consequent PGE2 formation in LPS-stimulated macrophages
- Author
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Goldsmith, Meir, Daka, Ala, Lamour, Nadia F., Mashiach, Roi, Glucksam, Yifat, Meijler, Michael M., Chalfant, Charles E., and Zor, Tsaffrir
- Subjects
- *
CERAMIDES , *MACROPHAGES , *PROSTAGLANDINS , *PHOSPHOLIPIDS , *GENE expression , *ARACHIDONIC acid , *LECITHIN - Abstract
Abstract: Prostaglandin E2 (PGE2) is an important mediator of the inflammatory response. Phospho-ceramide analogue-1 (PCERA-1), a synthetic phospholipid-like molecule, was previously reported to modulate pro- and anti-inflammatory cytokine production. We show here that PCERA-1 inhibited LPS-stimulated PGE2 production in RAW264.7 macrophages, without affecting COX-2 expression. Furthermore, PCERA-1 efficiently suppressed arachidonic acid (AA) release in response to LPS. The dephosphorylated derivative of PCERA-1, ceramide analogue-1 (CERA-1), mimicked the inhibitory effect of PCERA-1 on AA release and PGE2 production in macrophages. Inhibition of PGE2 production by CERA-1 was completely rescued by addition of exogenous AA. Importantly, PCERA-1 and ceramide-1-phosphate (C1P) stimulated the enzymatic activity of cPLA2α in an in vitro assay, whereas CERA-1 and ceramide inhibited both basal and C1P-stimulated cPLA2α activity. Collectively, these results indicate that CERA-1 suppresses AA release and subsequent PGE2 production in LPS-stimulated macrophages by direct interaction with cPLA2, and suggest that ceramide may similarly counteract C1P effect on cPLA2 activity in cells. The suppression of PGE2 production is suggested to contribute to the anti-inflammatory action of PCERA-1. [ABSTRACT FROM AUTHOR]
- Published
- 2011
- Full Text
- View/download PDF
14. Synergistic IL-10 induction by LPS and the ceramide-1-phosphate analog PCERA-1 is mediated by the cAMP and p38 MAP kinase pathways
- Author
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Goldsmith, Meir, Avni, Dorit, Ernst, Orna, Glucksam, Yifat, Levy-Rimler, Galit, Meijler, Michael M., and Zor, Tsaffrir
- Subjects
- *
INTERLEUKIN-10 , *ENDOTOXINS , *PHOSPHATES , *CERAMIDES , *ADENOSINE monophosphate , *MITOGEN-activated protein kinases , *CELLULAR signal transduction , *CYTOKINES , *GENETIC regulation - Abstract
Abstract: Expression of the anti-inflammatory cytokine IL-10 can be induced either by TLR agonists such as lipopolysaccharide (LPS), or by various endogenous stimuli, in particular those acting via a cAMP-dependent signaling pathway. We have previously reported that the synthetic phospho-ceramide analogue-1 (PCERA-1) increases cAMP level and subsequently down-regulates production of TNFα and up-regulates production of IL-10 in LPS-stimulated macrophages. The objective of this study was to determine the mechanism of activity of PCERA-1 and the role of cAMP in LPS-induced IL-10 production. We show here that PCERA-1 induces IL-10 production in synergism with various TLR agonists in mouse RAW264.7 macrophages. Cooperativity is evident both at the mRNA and protein levels. IL-10 production by LPS and PCERA-1 is mediated by the cAMP pathway and by the p38 MAP kinase. Phosphorylation of p38 is cooperatively accomplished by LPS and PCERA-1 or other cAMP inducers. Furthermore, the activity of PCERA-1 can be partially mimicked by a cell-permeable analog of cAMP, and blocked by the protein kinase A (PKA) inhibitor H89. Finally, in the absence of PCERA-1, the residual IL-10 induction by LPS depends on the basal cAMP level as it can be largely elevated by the phosphodiesterase (PDE)-4 inhibitor rolipram. Our results thus indicate that IL-10 induction by LPS critically depends on basal cAMP level, and that a co-stimulus by a TLR agonist and a cAMP-elevating agent results in synergistic PKA-dependent and p38-dependent IL-10 production. [Copyright &y& Elsevier]
- Published
- 2009
- Full Text
- View/download PDF
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