Identification of cyclophilin as a proinflammatory secretory product of lipopolysaccharide-activated macrophages.
We have isolated an 18-kDa peptide (designated sp18, for 18-kDa secreted protein) from the conditioned medium of lipopolysaccharide-stimulated RAW 264.7 murine macrophages. Purified sp18 had in vivo inflammatory activity and in vitro chemotactic activity for human peripheral blood polymorphonuclear leukocytes and monocytes. Surprisingly, N-terminal sequencing and tryptic mapping studies revealed that sp18 and cyclophilin, an intracellular protein that binds the immunosuppressive drug cyclosporin A, are highly homologous. The in vitro chemotactic activity of sp18 on monocytes was blocked by cyclosporin A but not by cyclosporin H, a structural analog of cyclosporin A that does not bind cyclophilin. Like purified porcine cyclophilin, mouse sp18 exhibited peptidyl-prolyl cis-trans isomerase activity. Medium conditioned by lipopolysaccharide-stimulated resident peritoneal exudate macrophages isolated from C57BL/6 mice contained substantially higher levels of sp18/cyclophilin than medium conditioned by nonstimulated macrophages. The observation that sp18/cyclophilin exhibits proinflammatory activity and is secreted by macrophages in response to endotoxin suggests that this protein may function as a cytokine, and invites the hypothesis that the immunosuppressive action of cyclosporin A results in part from interaction with an extracellular form of cyclophilin released as a mediator of immune and inflammatory functions.
Sherry
B
B
Laboratory of Medical Biochemistry, Rockefeller University, New York, NY 10021.
Yarlett
N
N
Strupp
A
A
Cerami
A
A
D000591:Amino Acid Isomerases / Q000378:metabolism / Q000494:pharmacology / Q000502:physiology*; D000595:Amino Acid Sequence; D000818:Animals; D002352:Carrier Proteins / Q000378:metabolism / Q000494:pharmacology / Q000502:physiology*; D002478:Cells, Cultured; D002634:Chemotaxis, Leukocyte / Q000187:drug effects; D003470:Culture Media; D003524:Cyclosporins / Q000494:pharmacology; D004926:Escherichia coli; D006801:Humans; D007249:Inflammation / Q000503:physiopathology*; D007963:Leukocytes, Mononuclear / Q000187:drug effects / Q000502:physiology*; D008070:Lipopolysaccharides / Q000633:toxicity*; D008262:Macrophage Activation; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009504:Neutrophils / Q000187:drug effects / Q000502:physiology*; D010449:Peptide Mapping; D019696:Peptidylprolyl Isomerase; D012689:Sequence Homology, Nucleic Acid; D013552:Swine; D014357:Trypsin
D002352:Carrier Proteins; D003470:Culture Media; D003524:Cyclosporins; D008070:Lipopolysaccharides; D014357:Trypsin; D000591:Amino Acid Isomerases; D019696:Peptidylprolyl Isomerase