The impact of platelet-activating factor (PAF)-like mediators on the functional activity of neutrophils: anti-inflammatory effects of human PAF-acetylhydrolase.
Platelet-activating factor (PAF) is a proinflammatory agent in infectious and inflammatory diseases, partly due to the activation of infiltrating phagocytes. PAF exerts its actions after binding to a monospecific PAF receptor (PAFR). The potent bioactivity is reflected by its ability to activate neutrophils at picomolar concentrations, as defined by changes in levels of intracellular Ca(2+) ([Ca(2+)](i)), and induction of chemotaxis and actin polymerization at nanomolar concentration. The role of PAF in neutrophil survival is, however, less well appreciated. In this study, the inhibitory effects of synthetic PAFR-antagonists on various neutrophil functions were compared with the effect of recombinant human plasma-derived PAF-acetylhydrolase (rPAF-AH), as an important enzyme for PAF degradation in blood and extracellular fluids. We found that endogenously produced PAF (-like) substances were involved in the spontaneous apoptosis of neutrophils. At concentrations of 8 microg/ml or higher than normal plasma levels, rPAF-AH prevented spontaneous neutrophil apoptosis (21 +/- 4% of surviving cells (mean +/- SD; control) versus 62 +/- 12% of surviving cells (mean +/- SD; rPAF-AH 20 microg/ml); P < 0.01), during overnight cultures of 15 h. This effect depended on intact enzymatic activity of rPAF-AH and was not due to the resulting product lyso-PAF. The anti-inflammatory activity of rPAF-AH toward neutrophils was substantiated by its inhibition of PAF-induced chemotaxis and changes in [Ca(2+)](i). In conclusion, the efficient and stable enzymatic activity of rPAF-AH over so many hours of coculture with neutrophils demonstrates the potential for its use in the many inflammatory processes in which PAF (-like) substances are believed to be involved.
Kuijpers
T W
TW
Emma Children's Hospital, Academic Medical Centre, University of Amsterdam, Amsterdam, The Netherlands. T.W.Kuijpers@amc.uva.nl
van den Berg
J M
JM
Tool
A T
AT
Roos
D
D
D043203:1-Alkyl-2-acetylglycerophosphocholine Esterase; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000494:pharmacology*; D017209:Apoptosis; D002118:Calcium / Q000032:analysis; D002470:Cell Survival; D002634:Chemotaxis, Leukocyte; D003600:Cytosol / Q000737:chemistry; D006801:Humans; D019255:NADPH Oxidases / Q000378:metabolism; D009504:Neutrophils / Q000187:drug effects*; D010741:Phospholipases A / Q000235:genetics / Q000494:pharmacology*; D010972:Platelet Activating Factor / Q000378:metabolism*; D010980:Platelet Membrane Glycoproteins / Q000037:antagonists & inhibitors*; D011956:Receptors, Cell Surface; D043562:Receptors, G-Protein-Coupled; D011994:Recombinant Proteins / Q000494:pharmacology
D000894:Anti-Inflammatory Agents, Non-Steroidal; D010972:Platelet Activating Factor; D010980:Platelet Membrane Glycoproteins; D011956:Receptors, Cell Surface; D043562:Receptors, G-Protein-Coupled; D011994:Recombinant Proteins; C039351:platelet activating factor receptor; D019255:NADPH Oxidases; D010741:Phospholipases A; D043203:1-Alkyl-2-acetylglycerophosphocholine Esterase; D002118:Calcium