Expression of the type-II phospholipase A2 in alveolar macrophages. Down-regulation by an inflammatory signal.
We have shown previously that guinea pig alveolar macrophages (AM) synthesize a secretory phospholipase A2 (PLA2) during in vitro incubation. Here, we report the molecular cloning of this enzyme and show that it has structural features closely related to all known mammalian type-II PLA2. The mRNA and PLA2 activity were undetectable in freshly collected AM, but their levels increased dramatically to reach maximal values after 16 h of culture. Thereafter, the PLA2 activity remained constant with a parallel secretion in the medium, in contrast to mRNA level which returned to near basal values after 32 h. Incubation of AM for 16 h with the inflammatory secretagogue peptide f-Met-Leu-Phe (fMLP) markedly reduced the PLA2 activity and mRNA levels. This inhibition was prevented by preexposure of AM to pertussis toxin, an inhibitor of G-protein. In contrast, when AM were first cultured for 16 h and then incubated with fMLP, no significant change was observed in their PLA2 activity. In conditions where the type-II PLA2 was completely abrogated by fMLP, the latter did not alter the lipopolysaccharide-induced accumulation of tumor necrosis factor alpha mRNA or the release of arachidonic acid induced by the subsequent addition of the calcium ionophore A23187. These studies show that the inflammatory peptide fMLP down-regulates the expression of the type-II PLA2 by AM through a process mediated by G-protein. A possible negative control of the type-II PLA2 expression during AM activation is suggested.
Vial
D
D
Unité de Pharmacologie Cellulaire, INSERM U285, Institut Pasteur, Paris, France.
Señorale-Pose
M
M
Havet
N
N
Molio
L
L
Vargaftig
B B
BB
Touqui
L
L
D000595:Amino Acid Sequence; D000818:Animals; D016718:Arachidonic Acid / Q000378:metabolism; D001483:Base Sequence; D003001:Cloning, Molecular; D015536:Down-Regulation; D019204:GTP-Binding Proteins / Q000502:physiology; D006168:Guinea Pigs; D007249:Inflammation / Q000378:metabolism*; D016676:Macrophages, Alveolar / Q000201:enzymology*; D008297:Male; D008969:Molecular Sequence Data; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000494:pharmacology; D010741:Phospholipases A / Q000096:biosynthesis* / Q000235:genetics; D054467:Phospholipases A2; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis
D014409:Tumor Necrosis Factor-alpha; D016718:Arachidonic Acid; D009240:N-Formylmethionine Leucyl-Phenylalanine; D010741:Phospholipases A; D054467:Phospholipases A2; D019204:GTP-Binding Proteins