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.

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.

• Authors: Dorit Avni, Meir Goldsmith, Orna Ernst, Roi Mashiach, Tove Tuntland, Michael M Meijler, Nathanael S Gray, Hugh Rosen, Tsaffrir Zor
• Journal: Immunology letters (Immunol Lett) Vol. 123 Issue 1 Pg. 1-8 (Mar 24 2009) ISSN: 1879-0542 [Electronic] Netherlands
• PMID: 19185589 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Ceramides
• Interleukin-12 Subunit p40
• Lipopolysaccharides
• PCERA-1
• Tumor Necrosis Factor-alpha
• Interleukin-10
• Interferon-gamma
• Dinoprostone

Topics

• Animals
• Cell Differentiation (drug effects, immunology)
• Ceramides (pharmacology)
• Dinoprostone (immunology, metabolism)
• Inflammation (immunology, metabolism)
• Interferon-gamma (pharmacology)
• Interleukin-10 (agonists, biosynthesis)
• Interleukin-12 Subunit p40 (antagonists & inhibitors, biosynthesis)
• Lipopolysaccharides (pharmacology)
• Macrophages (drug effects, immunology)
• Male
• Mice
• Mice, Inbred BALB C
• Monocytes (drug effects, immunology, metabolism)
• Tumor Necrosis Factor-alpha (antagonists & inhibitors, biosynthesis)