Experimental Arthritis Is Dependent on Mouse Mast Cell Protease-5

Richard L. Stevens; H. Patrick McNeil; Lislaine A. Wensing; Kichul Shin; G. William Wong; Philip M. Hansbro; Steven A. Krilis

doi:10.1074/jbc.M116.773416

Experimental Arthritis Is Dependent on Mouse Mast Cell Protease-5*

From the ^‡Department of Infectious Diseases, Immunology, and Sexual Health, St. George Hospital, and the St. George and Sutherland Clinical School, Faculty of Medicine, University of New South Wales, Sydney, New South Wales 2217, Australia,
the ^§Faculty of Health and Medicine, School of Biomedical Sciences and Pharmacy, University of Newcastle, Newcastle, New South Wales 2308, Australia,
the ^¶Faculty of Medicine and Health Sciences, Macquarie University, New South Wales 2109, Australia,
the ^‖Department of Immunology, Biomedical Sciences Institute, University of Sao Paulo, Sao Paulo 05508900, Brazil,
the ^**Division of Rheumatology, Seoul Metropolitan Government, Seoul National University (SMG-SNU), Boramae Medical Center, Seoul 07061, Korea,
the ^‡‡Department of Physiology, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205, and
the ^§§Centre for Asthma & Respiratory Disease, University of Newcastle and Hunter Medical Research Institute, Newcastle, New South Wales 2308, Australia

↵2 To whom correspondence should be addressed: Dept. of Infectious Diseases, Immunology, and Sexual Health, St. George Hospital, Research and Education Centre, Level 3, 4–10 South St., Kogarah, Sydney, NSW 2217, Australia. Tel.: 61-2-9113-2994; Fax: 61-2-9113-3981; E-mail: rstevens{at}richardstevensphd.org.

↵1 These authors contributed equally to this work.
Edited by George N. DeMartino

Abstract

The constitutive heparin⁺ (HP) mast cells (MCs) in mice express mouse MC protease (mMCP)-5 and carboxypeptidase A (mMC-CPA). The amino acid sequence of mMCP-5 is most similar to that of human chymase-1, as are the nucleotide sequences of their genes and transcripts. Using a homologous recombination approach, a C57BL/6 mouse line was created that possessed a disrupted mMCP-5 gene. The resulting mice were fertile and had no obvious developmental abnormality. Lack of mMCP-5 protein did not alter the granulation of the IL-3/IL-9-dependent mMCP-2⁺ MCs in the jejunal mucosa of Trichinella spiralis-infected mice. In contrast, the constitutive HP⁺ MCs in the tongues of mMCP-5-null mice were poorly granulated and lacked mMC-CPA protein. Bone marrow-derived MCs were readily developed from the transgenic mice using IL-3. Although these MCs contained high levels of mMC-CPA mRNA, they also lacked the latter exopeptidase. mMCP-5 protein is therefore needed to target translated mMC-CPA to the secretory granule along with HP-containing serglycin proteoglycans. Alternately, mMCP-5 is needed to protect mMC-CPA from autolysis in the cell's granules. Fibronectin was identified as a target of mMCP-5, and the exocytosis of mMCP-5 from the MCs in the mouse's peritoneal cavity resulted in the expression of metalloproteinase protease-9, which has been implicated in arthritis. In support of the latter finding, experimental arthritis was markedly reduced in mMCP-5-null mice relative to wild-type mice in two disease models.

Footnotes

↵* This work was supported in part by funds from the St. George and Sutherland Medical Research Foundation (to R. L. S. and S. K.), the Harvard Club of Australia Foundation (to R. L. S., S. K., and P. M. H.), the Australian Research Council (to P. M. H. and R. L. S.), and the University of Newcastle (to R. L. S.); National Institutes of Health Grants DK084171 (to G. W. W.) and AI059746 (to R. L. S.); a fellowship from the National Health and Medical Research Council of Australia (to P. M. H.); the Brawn Fellowship from the University of Newcastle (to P. M. H.); and a fellowship from Fundação de Amparo a Pesquisa do Estado de São Paulo-FAPESP (to L. A. W.). The authors declare that they have no conflicts of interest with the contents of this article. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.
↵ This article contains supplemental Tables S1 and S2.