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Thromb Haemost 2010; 103(03): 545-555
DOI: 10.1160/TH09-07-0441
Theme Issue Article
Schattauer GmbH

Neuroserpin, a thrombolytic serine protease inhibitor (serpin), blocks transplant vasculopathy with associated modification of T-helper cell subsets

Ganesh Munuswamy-Ramanujam
1   Division of Cardiovascular Medicine, Department of Medicine, University of Florida, Gainesville, Florida, USA
2   Division of Cardiovascular Medicine, Department of Molecular Genetics and Microbiology, University of Florida, Gainesville, Florida, USA
3   Robarts Research Institute, Department of Medicine, University of Western Ontario, London, Ontario, Canada
4   Department of Microbiology and Immunology, Department of Medicine, University of Western Ontario, London, Ontario, Canada
,
Erbin Dai
1   Division of Cardiovascular Medicine, Department of Medicine, University of Florida, Gainesville, Florida, USA
3   Robarts Research Institute, Department of Medicine, University of Western Ontario, London, Ontario, Canada
,
Liying Liu
1   Division of Cardiovascular Medicine, Department of Medicine, University of Florida, Gainesville, Florida, USA
3   Robarts Research Institute, Department of Medicine, University of Western Ontario, London, Ontario, Canada
,
Mira Shnabel
3   Robarts Research Institute, Department of Medicine, University of Western Ontario, London, Ontario, Canada
4   Department of Microbiology and Immunology, Department of Medicine, University of Western Ontario, London, Ontario, Canada
,
Yun Ming Sun
3   Robarts Research Institute, Department of Medicine, University of Western Ontario, London, Ontario, Canada
4   Department of Microbiology and Immunology, Department of Medicine, University of Western Ontario, London, Ontario, Canada
6   Viron Therapeutics, Inc., London, Ontario, Canada
,
Mee Bartee
1   Division of Cardiovascular Medicine, Department of Medicine, University of Florida, Gainesville, Florida, USA
2   Division of Cardiovascular Medicine, Department of Molecular Genetics and Microbiology, University of Florida, Gainesville, Florida, USA
,
David A. Lomas
7   Department of Medicine, Cambridge Institute for Medical Research, University of Cambridge, Cambridge, UK
,
Alexandra R. Lucas
1   Division of Cardiovascular Medicine, Department of Medicine, University of Florida, Gainesville, Florida, USA
2   Division of Cardiovascular Medicine, Department of Molecular Genetics and Microbiology, University of Florida, Gainesville, Florida, USA
3   Robarts Research Institute, Department of Medicine, University of Western Ontario, London, Ontario, Canada
4   Department of Microbiology and Immunology, Department of Medicine, University of Western Ontario, London, Ontario, Canada
5   Division of Cardiology, Department of Medicine, University of Western Ontario, London, Ontario, Canada
› Author Affiliations
Further Information

Publication History

Received: 07 July 2009

Accepted after major revision: 27 February 2009

Publication Date:
22 November 2017 (online)

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Summary

Thrombolytic serine proteases not only initiate fibrinolysis, but also are up-regulated in vascular disease and acute inflammatory responses. Although the serine protease inhibitor (serpin) plasminogen activator inhibitor-1 (PAI-1) is considered a main regulator of thrombolysis, PAI-1 is also associated with vascular inflammation. The role of other serpins that target thrombolytic proteases, PAI-2, PAI-3, and neuroserpin (NSP), in vascular inflammation is, however, less well defined. NSP is a mammalian serpin that, similar to PAI-1, inhibits urokinase- and tissue-type plasminogen activators (uPA and tPA, respectively) and has been most closely associated with the nervous system, with a demonstrated protective role after cerebral infarction in mouse models. However, the role of NSP in systemic arterial inflammation and plaque growth is not known. Serp-1 is a myxoma viral serpin that also inhibits tPA and uPA, as well as additionally inhibiting plasmin and factor Xa (fXa). Serp-1 has proven highly potent anti-inflammatory and anti-atherogenic activity. Here we assess the effects of NSP treatment on plaque growth and T-helper (Th) lymphocyte activity in a mouse aortic allograft transplant model, with comparison to Serp-1. NSP and Serp-1 both significantly reduced plaque growth and T-cell invasion. T-bet (a Th1 differentiation marker) was significantly reduced in transplanted aorta with associated reductions in Th1 and Th17, but not Th2, in splenocytes. NSP had additional Th modifying activity in non-transplanted mice. In summary, this is the first report that NSP possesses anti-inflammatory activity in systemic arteries, modifying Th cell responses and significantly reducing plaque growth in mouse aortic allografts.

Keywords

Serpin - neuroserpin - T-helper lymphocyte - transplant vasculopathy - atherosclerosis
 

  • References

  • 1 Weiss MJ, Madsen JC, Rosengard BR. et al. Mechanisms of chronic rejection in cardiothoracic transplantation. Front Biosci 2008; 13: 2980-2988.
    CrossrefPubMedSearch in Google Scholar
  • 2 Belperio JA, Weigt SS, Fishbein MC. et al. Chronic lung allograft rejection: mechanisms and therapy. Proc Am Thorac Soc 2009; 6: 108-121.
    CrossrefPubMedSearch in Google Scholar
  • 3 Cornell LD, Smith RN, Colvin RB. Kidney transplantation: mechanisms of rejection and acceptance. Annu Rev Pathol 2008; 3: 189-220.
    CrossrefPubMedSearch in Google Scholar
  • 4 Weis M, von Scheidt W. Cardiac allograft vasculopathy: a review. Circulation 1997; 96: 2069-2077.
    CrossrefPubMedSearch in Google Scholar
  • 5 Binder BR, Mihaly J, Prager GW. uPAR-uPA-PAI-1 interactions and signalling: a vascular biologist’s view. Thromb Haemost 2007; 97: 336-342.
    Thieme ConnectPubMedSearch in Google Scholar
  • 6 Mondino A, Blasi F. uPA and uPAR in fibrinolysis, immunity and pathology. Trends Immunol 2004; 25: 450-455.
    CrossrefPubMedSearch in Google Scholar
  • 7 Diebold I, Kraicun D, Bonello S. et al. The ‘PAI-1 paradox’ in vascular remodelling. Thromb Haemost 2008; 100: 984-991.
    Thieme ConnectPubMedSearch in Google Scholar
  • 8 Qian HS, Gu JM, Liu P. et al. Overexpression of PAI-1 prevents the development of abdominal aortic aneurysm in mice. Gene Ther 2008; 15: 224-232.
    CrossrefPubMedSearch in Google Scholar
  • 9 Nicholl SM, Roztocil E, Davies MG. Plasminogen activator system and vascular disease. Curr Vasc Pharmacol 2006; 4: 101-116.
    CrossrefPubMedSearch in Google Scholar
  • 10 Yepes M, Lawrence DA. Tissue-type plasminogen activator and neuroserpin: a well-balanced act in the nervous system?. Trends Cardiovasc Med 2004; 14: 173-180.
    CrossrefPubMedSearch in Google Scholar
  • 11 Miranda E, Lomas DA. Neuroserpin: a serpin to think about. Cell Mol Life Sci 2006; 63: 709-722.
    CrossrefPubMedSearch in Google Scholar
  • 12 Osterwalder T, Cinelli P, Baici A. et al. The axonally secreted serine proteinase inhibitor, neuroserpin, inhibits plasminogen activators and plasmin but not thrombin. J Biol Chem 1998; 273: 2312-2321.
    CrossrefPubMedSearch in Google Scholar
  • 13 Cinelli P, Madani R, Tsuzuki N. et al. Neuroserpin, a neuroprotective factor in focal ischemic stroke. Mol Cell Neurosci 2001; 18: 443-457.
    CrossrefPubMedSearch in Google Scholar
  • 14 Simonin Y, Charron Y, Sonderegger P. et al. An inhibitor of serine proteases, neuroserpin, acts as a neuroprotective agent in a mouse model of neurodegenerative disease. J Neurosci 2006; 26: 10614-10619.
    CrossrefPubMedSearch in Google Scholar
  • 15 Nash P, Whitty A, Handwerker J. et al. Inhibitory specificity of the anti-inflammatory myxoma virus serpin, SERP-1. J Biol Chem 1998; 33: 20982-20991.
    PubMedSearch in Google Scholar
  • 16 Dai E, Guan H, Liu L. et al. Serp-1, a viral anti-inflammatory serpin, regulates cellular serine proteinase and serpin responses to vascular injury. J Biol Chem 2003; 278: 18563-18572.
    CrossrefPubMedSearch in Google Scholar
  • 17 Bot I, von der Thüsen JH, Donners MM. et al. Serine protease inhibitor Serp-1 strongly impairs atherosclerotic lesion formation and induces a stable plaque phenotype in ApoE-/-mice. Circ Res 2003; 93: 464-471.
    CrossrefPubMedSearch in Google Scholar
  • 18 Bédard EL, Jiang J, Arp J. et al. Prevention of chronic renal allograft rejection by SERP-1 protein. Transplantation 2006; 81: 908-914.
    CrossrefPubMedSearch in Google Scholar
  • 19 Hausen B, Boeke K, Berry GJ. et al. Viral serine proteinase inhibitor (SERP-1) effectively decreases the incidence of graft vasculopathy in heterotopic heart allografts. Transplantation 2001; 72: 364-368.
    CrossrefPubMedSearch in Google Scholar
  • 20 Jiang J, Arp J, Kubelik D. et al. Induction of indefinite cardiac allograft survival correlates with toll-like receptor 2 and 4 downregulation after serine protease inhibitor-1 (Serp-1) treatment. Transplantation 2007; 84: 1158-1167.
    CrossrefPubMedSearch in Google Scholar
  • 21 Wang H, Jiang J, McFadden G. et al. Serp-1, a viral anti-inflammatory serpin, attenuates acute xenograft rejection in a rat-to-mouse cardiac transplantation model. Xenotransplant 2003; 5: 506.
    PubMedSearch in Google Scholar
  • 22 Lucas A, Dai E, Liu L. et al. Transplant vasculopathy: viral anti-inflammatory serpin regulation of atherogenesis. J Heart Lung Transplant 2000; 19: 1029-1038.
    CrossrefPubMedSearch in Google Scholar
  • 23 Richardson J, Viswanathan K, Lucas A. Serpins, the vasculature, and viral therapeutics. Front Biosci 2006; 1: 1042-1056.
    PubMedSearch in Google Scholar
  • 24 Zhou X. CD4+ T cells in atherosclerosis. Biomed Pharmacother 2003; 57: 287-291.
    CrossrefPubMedSearch in Google Scholar
  • 25 Zhou X, Robertson AK, Hjerpe C. et al. Adoptive transfer of CD4+ T cells reactive to modified low-density lipoprotein aggravates atherosclerosis. Arterioscler Thromb Vasc Biol 2006; 26: 864-870.
    CrossrefPubMedSearch in Google Scholar
  • 26 Robertson AK, Hansson GK. T cells in atherogenesis: for better or for worse?. Arterioscler Thromb Vasc Biol 2006; 26: 2421-2432.
    CrossrefPubMedSearch in Google Scholar
  • 27 Buono C, Binder CJ, Stavrakis G. et al. T-bet deficiency reduces atherosclerosis and alters plaque antigen-specific immune responses. Proc Natl Acad Sci USA 2005; 102: 1596-1601.
    CrossrefPubMedSearch in Google Scholar
  • 28 Kleemann R, Zadelaar S, Kooistra T. Cytokines and atherosclerosis: a comprehensive review of studies in mice. Cardiovasc Res 2008; 79: 360-376.
    CrossrefPubMedSearch in Google Scholar
  • 29 Huber SA, Sakkinen P, David C. et al. T helper-cell phenotype regulates atherosclerosis in mice under conditions of mild hypercholesterolemia. Circulation 2001; 103: 2610-2616.
    CrossrefPubMedSearch in Google Scholar
  • 30 Tesmer LA, Lundy SK, Sarkar S. Th17 cells in human disease. Immunol Rev 2008; 223: 87-113.
    CrossrefPubMedSearch in Google Scholar
  • 31 Afzali B, Lombardi G, Lechler RI. et al. The role of T helper 17 (Th17) and regulatory T cells (Treg) in human organ transplantation and autoimmune disease. Clin Exp Immunol 2007; 148: 32-46.
    CrossrefPubMedSearch in Google Scholar
  • 32 Cheng X, Yu X, Ding YJ. et al. The Th17/Treg imbalance in patients with acute coronary syndrome. Clin Immunol 2008; 127: 89-97.
    CrossrefPubMedSearch in Google Scholar
  • 33 Le Luduec JB, Condamine T, Louvet C. et al. An immunomodulatory role for follistatin-like 1 in heart allograft transplantation. Am J Transplant 2008; 8: 2297-2306.
    CrossrefPubMedSearch in Google Scholar
  • 34 Hsieh HG, Loong CC, Lui WY. et al. IL-17 expression as a possible predictive parameter for subclinical renal allograft rejection. Transpl Int 2001; 14: 287-298.
    CrossrefPubMedSearch in Google Scholar
  • 35 Lucas A, Liu L, Macen J. et al. Virus-encoded serine proteinase inhibitor SERP-1 inhibits atherosclerotic plaque development after balloon angioplasty. Circulation 1996; 94: 2890-2900.
    CrossrefPubMedSearch in Google Scholar
  • 36 Nash P, Barry M, Seet BT. et al. Post-translational modification of the myxoma-virus anti-inflammatory serpin SERP-1 by a virally encoded sialyltransferase. Biochem J 2000; 347: 375-382.
    CrossrefPubMedSearch in Google Scholar
  • 37 Dai E, Viswanathan K, Sun YM. et al. Identification of myxomaviral serpin reactive site loop sequences that regulate innate immune responses. J Biol Chem 2006; 281: 8041-8050.
    CrossrefPubMedSearch in Google Scholar
  • 38 Viswanathan K, Liu L, Vaziri S. et al. Myxoma viral serpin, Serp-1, a unique interceptor of coagulation and innate immune pathways. Thromb Haemost 2006; 95: 499-510.
    Thieme ConnectPubMedSearch in Google Scholar
  • 39 Mills KH. Induction, function and regulation of IL-17-producing T cells. Eur J Immunol 2008; 38: 2636-2649.
    CrossrefPubMedSearch in Google Scholar
  • 40 Iwakura Y, Nakae S, Saijo S. et al. The roles of IL-17A in inflammatory immune responses and host defense against pathogens. Immunol Rev 2008; 226: 57-79.
    CrossrefPubMedSearch in Google Scholar
  • 41 Petersen AM, Pedersen BK. The anti-inflammatory effect of exercise. J Appl Physiol 2005; 98: 1154-1162.
    CrossrefPubMedSearch in Google Scholar
  • 42 Levi M, Schouten M, van der Poll T. Sepsis, Coagulation, and Antithrombin: Old Lessons and New Insights. Semin Thromb Hemost 2008; 34: 742-746.
    Thieme ConnectPubMedSearch in Google Scholar
  • 43 Weber C, Zernecke A, Libby P. The multifaceted contributions of leukocyte subsets to atherosclerosis: lessons from mouse models. Nat Rev Immunol 2008; 8: 802-815.
    CrossrefPubMedSearch in Google Scholar
  • 44 Szabo SJ, Sullivan BM, Stemmann C. et al. Distinct effects of T-bet in Th1 lineage commitment and IFN-gamma production in CD4 and CD8 T cells. Science 2002; 295: 338-342.
    CrossrefPubMedSearch in Google Scholar
  • 45 Zheng W, Flavell RA. The transcription factor GATA-3 is necessary and sufficient for Th2 cytokine gene expression in CD4 T cells. Cell 1997; 89: 587-596.
    CrossrefPubMedSearch in Google Scholar
  • 46 Layh-Schmitt G, Colbert RA. The interleukin-23/interleukin-17 axis in spondyloarthritis. Curr Opin Rheumatol 2008; 20: 392-397.
    CrossrefPubMedSearch in Google Scholar
  • 47 Daftarian PM, Kumar A, Kryworuchko M. et al. IL-10 production is enhanced in human T cells by IL-12 and IL-6 and in monocytes by tumor necrosis factor-alpha. J Immunol 1996; 157: 12-20.
    PubMedSearch in Google Scholar
  • 48 Murata N, Los DA. Membrane fluidity and temperature perception. Plant Physiol 1997; 115: 875-879.
    CrossrefPubMedSearch in Google Scholar
  • 49 Vigh L, Maresca B, Harwood JL. Does the membrane’s physical state control the expression of heat shock and other genes?. Trends Biochem Sci 1998; 23: 369-374.
    CrossrefPubMedSearch in Google Scholar
  • 50 Zalai CV, Kolodziejczyk MD, Pilarski L. et al. Increased circulating monocyte activation in patients with unstable coronary syndromes. J Am Coll Cardiol 2001; 38: 1340-1348.
    CrossrefPubMedSearch in Google Scholar