Antisense knockdown of sphingosine kinase 1 in human macrophages inhibits C5a receptor-dependent signal transduction, Ca2+ signals, enzyme release, cytokine production, and chemotaxis.
The anaphylatoxin C5a is produced following the activation of the complement system and is associated with a variety of pathologies, including septic shock and adult respiratory distress syndrome, and with immune complex-dependent diseases such as rheumatoid arthritis. C5a has been shown to regulate inflammatory functions by interacting with its receptor, C5aR, which belong to the rhodopsin family of seven-transmembrane GPCRs. However, the intracellular signaling pathways triggered by C5aR on immune-effector cells are not well understood. In this report we present data showing that, in human monocyte-derived macrophages, C5aR uses the intracellular signaling molecule sphingosine kinase (SPHK)1 to trigger various physiological responses. Our data show that C5a rapidly stimulates the generation of sphingosine-1-phosphate, SPHK activity, and membrane translocation of SPHK1. Using an antisense oligonucleotide against SPHK1, we show that knockdown of SPHK1 abolishes the C5a-triggered intracellular Ca(2+) signals, degranulation, cytokine generation, and chemotaxis. Our study shows for the first time that SPHK1 not only plays a key role in the generation and release of proinflammatory mediators triggered by anaphylatoxins from human macrophages but is also involved in the process of immune cell motility, thus pointing out SPHK1 as a potential therapeutic target for the treatment of inflammatory and autoimmune diseases.
Melendez
Alirio J
AJ
Department of Physiology, National University of Singapore, Singapore. phsmraj@nus.edu.sg
Ibrahim
Farazeela Bte Mohd
FB
D020013:Calcium Signaling / Q000187:drug effects; D002478:Cells, Cultured / Q000187:drug effects / Q000201:enzymology; D002633:Chemotaxis / Q000187:drug effects*; D015936:Complement C5a / Q000494:pharmacology; D016207:Cytokines / Q000096:biosynthesis* / Q000378:metabolism; D003594:Cytoplasmic Granules / Q000201:enzymology / Q000378:metabolism; D006801:Humans; D015850:Interleukin-6 / Q000096:biosynthesis / Q000378:metabolism; D016209:Interleukin-8 / Q000096:biosynthesis / Q000378:metabolism; D008246:Lysophospholipids / Q000378:metabolism; D008264:Macrophages / Q000187:drug effects / Q000201:enzymology*; D020319:Oligodeoxyribonucleotides, Antisense / Q000494:pharmacology*; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000037:antagonists & inhibitors / Q000235:genetics / Q000502:physiology*; D011493:Protein Kinase C / Q000378:metabolism; D044087:Receptor, Anaphylatoxin C5a / Q000037:antagonists & inhibitors*; D015398:Signal Transduction / Q000187:drug effects*; D013110:Sphingosine / Q000031:analogs & derivatives / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis / Q000378:metabolism; D010738:Type C Phospholipases / Q000378:metabolism; D001619:beta-N-Acetylhexosaminidases / Q000378:metabolism*; D000099157:Sphingosine Kinase
D015936:Complement C5a; D016207:Cytokines; D015850:Interleukin-6; D016209:Interleukin-8; D008246:Lysophospholipids; D020319:Oligodeoxyribonucleotides, Antisense; D017853:Phosphotransferases (Alcohol Group Acceptor); D011493:Protein Kinase C; D044087:Receptor, Anaphylatoxin C5a; D013110:Sphingosine; D014409:Tumor Necrosis Factor-alpha; D010738:Type C Phospholipases; D001619:beta-N-Acetylhexosaminidases; D000099157:Sphingosine Kinase; C060506:sphingosine 1-phosphate