title: Human mast cell tryptase: a stimulus of microvascular leakage and mast cell activation.
abstract: We have investigated the potential of tryptase to stimulate an increase in microvascular permeability following injection into the skin of guinea pigs. Tryptase was isolated from high salt extracts of human lung tissue by octyl-agarose and heparin-agarose chromatography. Injection of purified tryptase (2.5 ng-2.5 microg/site) into the skin of guinea pigs which had been injected intravenously with Evans blue dye provoked a dose-dependent increase in microvascular permeability. The skin reactions elicited by tryptase were apparent up to 80 min following injection, while histamine-induced microvascular leakage resolved completely by 40 min. Heat-inactivation of tryptase, or preincubating the proteinase with certain proteinase inhibitors, significantly reduced the extent of microvascular leakage, suggesting dependency on an intact catalytic site. No evidence was found for a synergistic or antagonistic interaction between tryptase (2.5 ng-2.5 microg/site) and histamine (1-10 microg/site) when these mast cell products were injected together. Addition of heparin to tryptase (10:1; w/w) prior to injection was without effect on tryptase-induced microvascular leakage. Pretreatment of guinea pigs with a combination of the histamine H1 receptor antagonist pyrilamine and the histamine H2 receptor antagonist cimetidine (both 10 mg/kg), partially abolished tryptase-induced microvascular leakage as well as attenuating the reaction to histamine. Reasoning that the microvascular leakage induced by tryptase is likely to involve the release of histamine, we investigated the ability of tryptase to stimulate histamine release from dispersed guinea-pig skin and lung cells in vitro. Tryptase was found to induce concentration-dependent histamine release from both sources of tissue. Mast cell activation stimulated by tryptase in vitro was inhibited by heat treating the enzyme or by addition of proteinase inhibitors, suggesting a requirement for an intact catalytic site. Histamine release was inhibited also by preincubating cells with the metabolic inhibitors antimycin A and 2-deoxy-D-glucose indicating that the mechanism was energy-requiring and non-cytotoxic. We conclude that human mast cell tryptase may be a potent stimulus of microvascular leakage. The activation of mast cells by this proteinase may represent an amplification process in allergic inflammation.
authors.affiliation: Immunopharmacology Group, Southampton General Hospital, UK.
authors.forename: S
authors.initials: S
authors.lastname: He
authors.forename: A F
authors.initials: AF
authors.lastname: Walls
mesh_terms: D000818:Animals; D002199:Capillary Permeability / Q000187:drug effects* / Q000502:physiology; D002846:Chromatography, Affinity; D053818:Chymases; D002927:Cimetidine / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004347:Drug Interactions; D005070:Evans Blue / Q000008:administration & dosage / Q000378:metabolism; D005119:Extravasation of Diagnostic and Therapeutic Materials; D006168:Guinea Pigs; D006493:Heparin / Q000494:pharmacology; D006632:Histamine / Q000633:toxicity; D006634:Histamine H1 Antagonists / Q000494:pharmacology; D006635:Histamine H2 Antagonists / Q000494:pharmacology; D006358:Hot Temperature; D006801:Humans; D018836:Inflammation Mediators / Q000008:administration & dosage / Q000378:metabolism / Q000494:pharmacology*; D007275:Injections, Intravenous; D008168:Lung / Q000378:metabolism; D008407:Mast Cells / Q000166:cytology / Q000187:drug effects / Q000201:enzymology*; D011480:Protease Inhibitors / Q000494:pharmacology; D011738:Pyrilamine / Q000494:pharmacology; D012697:Serine Endopeptidases / Q000008:administration & dosage / Q000378:metabolism / Q000494:pharmacology*; D012867:Skin / Q000098:blood supply / Q000187:drug effects / Q000378:metabolism; D053802:Tryptases
chemical_list: D006634:Histamine H1 Antagonists; D006635:Histamine H2 Antagonists; D018836:Inflammation Mediators; D011480:Protease Inhibitors; D005070:Evans Blue; D002927:Cimetidine; D006632:Histamine; D006493:Heparin; D012697:Serine Endopeptidases; C083294:chymase 2; D053818:Chymases; D053802:Tryptases; D011738:Pyrilamine