title: The granzyme B inhibitor, PI-9, is present in endothelial and mesothelial cells, suggesting that it protects bystander cells during immune responses.
abstract: Proteinase inhibitor 9 (PI-9) is a 42-kDa human intracellular serpin present in cytotoxic lymphocytes (CLs). PI-9 is an extremely efficient inhibitor of the pro-apoptotic CL granule proteinase granzyme B and is thought to function in the cytosol of CLs to protect against apoptosis induced by endogenously expressed or released granzyme B, particularly during target cell killing. Here we show by immunohistochemistry that PI-9 is also present in endothelial cells, in every tissue examined. Cultured endothelial cells express functional PI-9 (as assessed by binding to recombinant granzyme B) localized to the cytoplasm and nucleus. Immunohistochemistry also showed PI-9 in mesothelial cells, and this was confirmed by analysis of primary cells cultured from pleural and serous effusions. Granzyme B expression was not detected in either endothelial or mesothelial cells. In both cell types, PI-9 is up-regulated at the mRNA and protein level by exposure to the phorbol ester PMA, consistent with a response to inflammatory stimuli. We postulate that PI-9 is present in these lining cell types to protect against misdirected, free granzyme B released during a local immune response.
authors.affiliation: Department of Biochemistry and Molecular Biology, Monash University, Victoria, 3800, Australia.
authors.forename: M S
authors.initials: MS
authors.lastname: Buzza
authors.forename: C E
authors.initials: CE
authors.lastname: Hirst
authors.forename: C H
authors.initials: CH
authors.lastname: Bird
authors.forename: P
authors.initials: P
authors.lastname: Hosking
authors.forename: J
authors.initials: J
authors.lastname: McKendrick
authors.forename: P I
authors.initials: PI
authors.lastname: Bird
mesh_terms: D001202:Ascitic Fluid / Q000378:metabolism; D002460:Cell Line; D002461:Cell Line, Transformed; D002478:Cells, Cultured; D004727:Endothelium / Q000166:cytology / Q000187:drug effects / Q000276:immunology*; D004847:Epithelial Cells / Q000166:cytology / Q000187:drug effects; D004848:Epithelium / Q000276:immunology*; D053804:Granzymes; D006801:Humans; D007150:Immunohistochemistry; D007249:Inflammation / Q000378:metabolism; D012333:RNA, Messenger / Q000096:biosynthesis; D012697:Serine Endopeptidases / Q000378:metabolism*; D015842:Serine Proteinase Inhibitors / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D015843:Serpins / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D015854:Up-Regulation
chemical_list: D012333:RNA, Messenger; C102316:SERPINB9 protein, human; D015842:Serine Proteinase Inhibitors; D015843:Serpins; C509213:GZMB protein, human; D053804:Granzymes; D012697:Serine Endopeptidases; D013755:Tetradecanoylphorbol Acetate