ABSTRACT
When mouse bone marrow-derived macrophages were stimulated with serum amyloid A (SAA), which is a major acute-phase protein, there was strong inhibition of osteoclast formation induced by the receptor activator of nuclear factor kappaB ligand. SAA not only markedly blocked the expression of several osteoclast-associated genes (TNF receptor-associated factor 6 and osteoclast-associated receptor) but also strongly induced the expression of negative regulators (MafB and interferon regulatory factor 8). Moreover, SAA decreased c-fms expression on the cell surface via shedding of the c-fms extracellular domain. SAA also restrained the fusion of osteoclast precursors by blocking intracellular ATP release. This inhibitory response of SAA is not mediated by the well-known SAA receptors (formyl peptide receptor 2, Toll-like receptor 2 (TLR2) or TLR4). These findings provide insight into a novel inhibitory role of SAA in osteoclastogenesis and suggest that SAA is an important endogenous modulator that regulates bone homeostasis.
Subject(s)
Animals , Humans , Mice , Adenosine Triphosphate/metabolism , Cell Differentiation , Cell Line , Gene Expression Regulation, Developmental , Macrophages/cytology , Osteoclasts/cytology , RANK Ligand/metabolism , Receptor, Macrophage Colony-Stimulating Factor/genetics , Receptors, Formyl Peptide/metabolism , Serum Amyloid A Protein/metabolism , Toll-Like Receptor 2/metabolism , Toll-Like Receptor 4/metabolismABSTRACT
Serum amyloid A (SAA) has been regarded as an important mediator of inflammatory responses. The effect of several formyl peptide receptor-like 1 (FPRL1) ligands on the production of IL-8 by SAA was investigated in human neutrophils. Among the ligands tested, LL-37 was found to specifically inhibit SAA-induced IL-8 production in transcriptional and post-transcriptional levels. Since SAA stimulated IL-8 production via ERK and p38 MAPK in human neutrophils, we tested the effect of LL-37 on SAA induction for these two MAPKs. LL-37 caused a dramatic inhibition of ERK and p38 MAPK activity, which is induced by SAA. LL-37 was also found to inhibit SAA-stimulated neutrophil chemotactic migration. Further, the LL-37-induced inhibitory effect was mediated by FPRL1. Our findings indicate that LL-37 is expected to be useful in the inhibition of SAA signaling and for the development of drugs against SAA-related inflammatory diseases.
Subject(s)
Animals , Humans , Rats , Antimicrobial Cationic Peptides/pharmacology , Cell Line, Tumor , Cell Movement , Chemotaxis, Leukocyte , Interleukin-8/biosynthesis , MAP Kinase Kinase Kinases/metabolism , Neutrophils/drug effects , Proto-Oncogene Proteins/metabolism , Receptors, Formyl Peptide/metabolism , Receptors, Lipoxin/metabolism , Serum Amyloid A Protein/antagonists & inhibitors , Signal Transduction , Transcription, GeneticABSTRACT
Serum amyloid A (SAA) has been regarded as an important mediator of inflammatory responses. The effect of several formyl peptide receptor-like 1 (FPRL1) ligands on the production of IL-8 by SAA was investigated in human neutrophils. Among the ligands tested, LL-37 was found to specifically inhibit SAA-induced IL-8 production in transcriptional and post-transcriptional levels. Since SAA stimulated IL-8 production via ERK and p38 MAPK in human neutrophils, we tested the effect of LL-37 on SAA induction for these two MAPKs. LL-37 caused a dramatic inhibition of ERK and p38 MAPK activity, which is induced by SAA. LL-37 was also found to inhibit SAA-stimulated neutrophil chemotactic migration. Further, the LL-37-induced inhibitory effect was mediated by FPRL1. Our findings indicate that LL-37 is expected to be useful in the inhibition of SAA signaling and for the development of drugs against SAA-related inflammatory diseases.
Subject(s)
Animals , Humans , Rats , Antimicrobial Cationic Peptides/pharmacology , Cell Line, Tumor , Cell Movement , Chemotaxis, Leukocyte , Interleukin-8/biosynthesis , MAP Kinase Kinase Kinases/metabolism , Neutrophils/drug effects , Proto-Oncogene Proteins/metabolism , Receptors, Formyl Peptide/metabolism , Receptors, Lipoxin/metabolism , Serum Amyloid A Protein/antagonists & inhibitors , Signal Transduction , Transcription, GeneticABSTRACT
In this study, we observed that lysophosphatidylglycerol (LPG) completely inhibited a formyl peptide receptor like-1 (FPRL1) agonist (MMK-1)-stimulated chemotactic migration in human phagocytes, such as neutrophils and monocytes. LPG also dramatically inhibited IL-1beta production by another FPRL1 agonist serum amyloid A (SAA) in human phagocytes. However, LPG itself induced intracellular calcium increase and superoxide anion production in human phagocytes. Keeping in mind that phagocytes migration and IL-1beta production by FPRL1 are important for the induction of inflammatory response, our data suggest that LPG can be regarded as a useful material for the modulation of inflammatory response induced by FPRL1 activation.
Subject(s)
Humans , Chemotaxis, Leukocyte/drug effects , Interleukin-1beta/biosynthesis , Lysophospholipids/pharmacology , Monocytes/drug effects , Neutrophils/drug effects , Peptides/metabolism , Phagocytes/drug effects , Receptors, Formyl Peptide/metabolism , Receptors, Lipoxin/metabolism , Serum Amyloid A Protein/metabolismABSTRACT
Dendritic cells (DCs) play a key role in activating the immune response against invading pathogens as well as dying cells or tumors. Although the immune response can be initiated by the phagocytic activity by DCs, the molecular mechanism involved in this process has not been fully investigated. Trp-Lys-Tyr-Met-Val-Met-NH2 (WKYMVM) stimulates the activation of phospholipase D (PLD) via Ca2+ increase and protein kinase C activation in mouse DC cell line, DC2.4. WKYMVM stimulates the phagocytic activity, which is inhibited in the presence of N-butanol but not t-butanol in DC2.4 cells. Furthermore, the addition of phosphatidic acid, an enzymatic product of PLD activity, enhanced the phagocytic activity in DC2.4 cells. Since at least two of formyl peptide receptor (FPR) family (FPR1 and FPR2) are expressed in DC2.4 as well as in mouse bone marrow-derived dendritic cells, this study suggests that the activation of FPR family by WKYMVM stimulates the PLD activity resulting in phagocytic activity in DC2.4 cells.