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(Arteriosclerosis, Thrombosis, and Vascular Biology. 2004;24:2245.)
© 2004 American Heart Association, Inc.

Vascular Biology

Granzyme B Induces Smooth Muscle Cell Apoptosis in the Absence of Perforin

Involvement of Extracellular Matrix Degradation

Jonathan C. Choy; Vivian H.Y. Hung; Arwen L. Hunter; Paul K. Cheung; Bruce Motyka; Ing Swie Goping; Tracy Sawchuk; R. Chris Bleackley; Thomas J. Podor; Bruce M. McManus; David J. Granville

From The James Hogg iCAPTURE Centre for Cardiovascular and Pulmonary Research (J.C.C., V.H.Y.H., A.L.H., P.K.C., T.J.P., B.M.M., D.J.G.), Department of Pathology and Laboratory Medicine, University of British Columbia, Vancouver, British Columbia, Canada; Department of Biochemistry (B.M., I.S.G., T.S., R.C.B.), University of Alberta, Edmonton, Alberta, Canada.

Correspondence to David J. Granville, PhD, The iCAPTURE Centre, Room 166, Burrard Building, St. Paul’s Hospital, 1081 Burrard St, Vancouver, BC, V6Z 1Y6 Canada. E-mail dgranville{at}mrl.ubc.ca

Objective— T cell-induced cytotoxicity, of which granzyme B is a key mediator, is believed to contribute to the pathogenesis of inflammatory vascular diseases. In this report, we investigate the mechanism of granzyme B-induced smooth muscle cell (SMC) death.

Methods and Results— The addition of purified granzyme B alone to cultured SMCs caused a significant reduction in cell viability. Chromatin condensation, phosphatidylserine externalization, and membrane blebbing were observed, indicating that the mechanism of granzyme B-induced SMC death was through apoptosis. Activated splenocytes from perforin-knockout mice induced SMC death through a granzyme B-mediated pathway. Inhibition of the proteolytic activities of caspases and granzyme B prevented granzyme B-induced SMC death, whereas attenuation of granzyme B internalization with mannose-6-phosphate (M6P) did not. Further, granzyme B induced the cleavage of several SMC extracellular proteins, including fibronectin, and reduced focal adhesion kinase phosphorylation.

Conclusions— These results indicate that granzyme B can induce apoptosis of SMCs in the absence of perforin by cleaving extracellular proteins, such as fibronectin.

We studied the mechanisms of granzyme B-induced smooth muscle cell (SMC) death. In the absence of perforin, granzyme B induced SMC apoptosis. Internalization of granzyme B was not required for SMC death, and this protease induced the cleavage of several extracellular proteins, including fibronectin.

Key Words: granzyme B • perforin • smooth muscle cell • apoptosis • extracellular

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Ken A. Lindstedt and Petri T. Kovanen
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