Skip to main content

Microarray analysis reveals the role of matrix metalloproteinases in mouse experimental autoimmune myocarditis induced by cardiac myosin peptides


Autoimmune myocarditis develops after the presentation of heart-specific antigens to autoaggressive CD4+ T cells and after inflammation has infiltrated the tissues. To shed light on global changes in the gene expression of autoimmune myocarditis and to gain further insight into the molecular mechanisms underlying the genesis of myocarditis, we conducted a comprehensive microarray analysis of mRNA using an experimental mouse autoimmune myocarditis model via immunization with α-myosin heavy chain-derived peptides. Of over 39,000 transcripts on a high density oligonucleotide microarray, 466 were under-expressed and 241 over-expressed by ≥ 1.5-fold compared with the controls in BALB/C mouse with autoimmune myocarditis. In this paper, we list the top 50 up-regulated genes related to the immune response. These altered genes encode for leukocyte-specific markers and receptors, the histocompatibility complex, cytokines/receptors, chemokines/receptors, adhesion molecules, components of the complement cascade, and signal transduction-related molecules. Interestingly, matrix metalloproteinases (MMPs) such as MMP-3 and MMP-9 were up-regulated, as further revealed by the reverse transcriptase-polymerase chain reaction (RT-PCR) and immunohistochemistry assays. This indicates that MMPs may act as major regulators of the cytokine profile. Together, these findings provide new insight into the molecular events associated with the mechanism of the autoimmune genesis of myocarditis.



complete Freund’s adjuvant


experimental autoimmune myocarditis


extracellular matrix


monocyte chemoattractant protein


major histocompatibility complex


macrophage inflammatory protein


matrix metalloproteinase


phosphate buffered solution


reverse transcriptase-polymerase chain reaction


tissue inhibitors of metalloproteinases


  1. Caforio, A.L., Mahon, N.J. and McKenna, W.J. Cardiac autoantibodies to myosin and other heart-specific autoantigens in myocarditis and dilated cardiomyopathy. Autoimmunity 34 (2001) 199–204.

    CAS  PubMed  Google Scholar 

  2. Neumann, D.A., Rose, N.R., Ansari, A.A. and Herskowitz, A. Induction of multiple heart autoantibodies in mice with coxsackievirus B3-and cardiac myosin-induced autoimmune myocarditis. J. Immunol. 152 (1994) 343–350.

    CAS  PubMed  Google Scholar 

  3. Kodama, M., Matsumoto, Y., Fujiwara, M., Masani, F. and Shibata, A. A novel experimental model of giant cell myocarditis induced in rats by immunization with cardiac myosin fraction. Clin. Immunol. Immunopathol. 57 (1990) 250–262.

    Article  CAS  PubMed  Google Scholar 

  4. Neu N., Rose, N.R., Beisel, K.W., Herskowitz, A., Gurri, G.G. and Craig, S.W. Cardiac myosin induces myocarditis in genetically predisposed mice. J. Immunol. 139 (1987) 3630–3636.

    CAS  PubMed  Google Scholar 

  5. Pummerer, C.L., Luze, K., Grässl, G., Bachmaier, K., Offner, F., Burrell, S.K., Lenz, D.M., Zamborelli, T.J., Penninger J.M. and Neu, N. Identification of Cardiac Myosin Peptides Capable of Inducing Autoimmune Myocarditis in BALB/c Mice. J. Clin. Invest. 97 (1996) 2057–2062.

    CAS  PubMed  Google Scholar 

  6. Eriksson, U., Ricci, R., Hunziker, L., Kurrer, M.O., Oudit, G.Y., Watts, T.H., Sonderegger, I., Bachmaier, K., Kopf, M. and Penninger, J.M. Dendritic cell-induced autoimmune heart failure requires cooperation between adaptive and innate immunity. Nat. Med. 9 (2003) 1484–1490.

    Article  CAS  PubMed  Google Scholar 

  7. Pummerer, C., Berger, P., Fruhwirth, M., Ofner, C. and Neu, N. Cellular infiltrate, major histocompatibility antigen expression and immunopathogenic mechanisms in cardiac myosin-induced myocarditis. Lab. Invest. 65 (1991) 538–547.

    CAS  PubMed  Google Scholar 

  8. Smith, S.C. and Allen, P.M. Myosin-induced acute myocarditis is a T cell-mediated disease. J. Immunol. 147 (1991) 2141–2147.

    CAS  PubMed  Google Scholar 

  9. Izumi, T., Suzuki, K., Saeki, M., Ookura, Y., Hirono, S., Inomata, T., Hanawa, H. and Kodama, M. An ultrastructural study on experimental autoimmune myocarditis with special reference to effector cells. Eur. Heart J. 16(suppl O) (1995) 75–77.

    PubMed  Google Scholar 

  10. Wang, Y., Afanasyeva, M., Hill, S.L. and Rose, N.R. Characterization of murine autoimmune myocarditis induced by self and foreign cardiac myosin. Autoimmunity 31 (1999) 151–162.

    Article  CAS  PubMed  Google Scholar 

  11. Lock, C., Hermans, G., Pedotti, R., Brendolan, A., Schadt, E., Garren, H., Langer-Gould, A., Strober, S., Cannella, B., Allard, J., Klonowski, P., Austin, A., Lad N., Kaminski, N., Galli, S.J., Oksenberg, J.R., Raine, C.S., Heller, R. and Steinman, L. Gene-microarray analysis of multiple sclerosis lesions yields new targets validated in autoimmune encephalomyelitis. Nat. Med. 8 (2002) 500–508.

    Article  CAS  PubMed  Google Scholar 

  12. Matejuk, A., Dwyer, J., Zamora, A., Vandenbark, A.A. and Offner, H. Evaluation of the effects of 17beta-estradiol (17beta-e2) on gene expression in experimental autoimmune encephalomyelitis using DNA microarray. Endocrinology 143 (2002) 313–319.

    Article  CAS  PubMed  Google Scholar 

  13. Aronow, B.J., Toyokawa, T., Canning, A., Haghighi, K., Delling, U., Kranias, E., Molkentin, J.D. and Dorn, G.W. Divergent transcriptional responses to independent genetic causes of cardiac hypertrophy. Physiol. Genomic. 6 (2001) 19–28.

    CAS  Google Scholar 

  14. Stanton L.W., Garrard L.J., Damm D., Garrick B.L., Lam A., Kapoun A.M., Zheng, Q., Protter, A.A., Schreiner, G.F. and White T. Altered patterns of gene expression in response to myocardial infarction. Circ. Res. 86 (2000) 939–945.

    CAS  PubMed  Google Scholar 

  15. Cihakova, D., Sharma, R.B., Fairweather, D., Afanasyeva, M. and Rose, N.R. Animal Models for Autoimmune Myocarditis and Autoimmune Thyroiditis. in Autoimmunity: methods and protocols (Perl, A., Ed.), New Jersey, Humana Press, 2004 pp 175–193.

    Google Scholar 

  16. Ducharme, A., Frantz, S., Aikawa, M., Rabkin, E., Lindsey, M., Rohde, L.E., Schoen, F.J., Kelly, R.A., Werb, Z., Libby, P. and Lee, R.T. Targeted deletion of matrix metalloproteinase-9 attenuates left ventricular enlargement and collagen accumulation after experimental myocardial infarction. J. Clin. Invest. 106 (2000) 55–62.

    CAS  PubMed  Google Scholar 

  17. Kim, H.E., Dalal, S.S., Young, E. Legato, M.J., Weisfeldt, M.L. and D’Armiento, J. Disruption of the myocardial extracellular matrix leads to cardiac dysfunction. J. Clin. Invest. 106 (2000) 857–866.

    Article  CAS  PubMed  Google Scholar 

  18. Creemers, E.E.J.M., Cleutjens, J.P.M., Smits, J.F.M. and Daemen, M.J.A.P. Matrix metalloproteinase inhibition after myocardial infarction: A new approach to prevent heart failure? Circ. Res. 89 (2001) 201–210.

    Article  CAS  PubMed  Google Scholar 

  19. Kodama, M., Zhang, S., Hanawa, H. and Shibata, A. Immunohistochemical characterization of infiltrating mononuclear cells in the rat heart with experimental autoimmune giant cell myocarditis. Clin. Exp. Immunol. 90 (1992) 330–335.

    Article  CAS  PubMed  Google Scholar 

  20. Hanawa, H., Tsuchida, M., Matsumoto, Y., Watanabe, H., Abo, T., Sekikawa, H., Kodama, M., Zhang, S., Lzumi, T. and Shibata, A. Characterization of T cells infiltrating the heart in rats with experimental autoimmune myocarditis. Their similarity to extrathymic T cells in mice and the site of proliferation. J. Immunol. 150 (1993) 5682–5695.

    CAS  PubMed  Google Scholar 

  21. Kaya, Z., Afanasyeva, M., Wang, Y., Dohmen, K.M., Schlichting, J., Tretter, T., Delisa, F., Holers V.M. and Rose, N.R. Contribution of the innate immune system to autoimmune myocarditis: a role for complement. Nat. Immunol. 2 (2001) 739–745.

    Article  CAS  PubMed  Google Scholar 

  22. Fuse, K., Kodama, M., Hanawa, H., Okura, Y., Ito, M., Shiono, T., Kato, K., Watanabe, K. and Aizawa, Y. Enhanced expression and production of monocyte chemoattractant protein-1 in myocarditis. Clin. Exp. Immunol. 124 (2001) 346–352.

    Article  CAS  PubMed  Google Scholar 

  23. Kurrer, M.O., Kopf, M., Penninger, J.M. and Eriksson, U. Cytokines that regulate autoimmune myocarditis. Swiss. Med. Wkly. 132 (2002) 408–413.

    CAS  PubMed  Google Scholar 

  24. Göser, S., Öttl, R., Brodner, A., Dengler, T.J., Torzewski, J., Egashira, K., Rose, N.R., Katus, H.A. and Kaya, Z. Critical role for monocyte chemoattractant protein-1 and macrophage inflammatory protein-1a in induction of experimental autoimmune myocarditis and effective antimonocyte chemoattractant protein-1 gene therapy. Circulation 112 (2005) 3400–3407.

    Article  PubMed  CAS  Google Scholar 

  25. Rouet-Benzineb, P., Buhler, J.M., Dreyfus, P., Delcourt, A., Dorent, R., Perennec, J., Crozatier, B., and Harf, A. and Lafuma, C. Altered balance between matrix gelatinases (MMP-2 and MMP-9) and their tissue inhibitors in human dilated cardiomyopathy: potential role of MMP-9 in myosin heavy chain degradation. Eur. J. Heart. Fail. 1 (1999) 337–352.

    Article  CAS  PubMed  Google Scholar 

  26. Thomas, C.V., Coker, M.L., Zellner, J.L., Handy, J.R., Crumbley, A.J. 3rd and Spinale, F.G. Increased matrix metalloproteinase activity and selective upregulation in LV myocardium from patients with end-stage dilated cardiomyopathy. Circulation 97 (1998) 1708–1715.

    CAS  PubMed  Google Scholar 

  27. Van Den Steen, P.E., Wuyts, A., Husson, S.J., Proost, P., Van, D.J. and Opdenakker, G. Gelatinase B/MMP-9 and neutrophil collagenase/MMP-8 process the chemokines human GCP-2/CXCL6, ENA-78/CXCL5 and mouse GCP-2/LIX and modulate their physiological activities. Eur. J. Biochem. 270 (2003) 3739–3749.

    Article  CAS  Google Scholar 

  28. Vreugdenhil, G.R., Wijnands, P.G., Netea, M.G., van der Meer, J.W., Melchers, W.J. and Galama, J.M. Enterovirus-induced production of proinflammatory and T-helper cytokines by human leukocytes. Cytokine 12 (2000) 1793–1796.

    Article  CAS  PubMed  Google Scholar 

  29. Sternlicht, M.D. and Werb, Z. How matrix metalloproteinases regulate cell behavior. Annu. Rev. Cell. Dev. Biol. 17 (2001) 463–516.

    Article  CAS  PubMed  Google Scholar 

  30. Gaertner, R., Jacob, M.P., Prunier, F., Angles-Cano, E., Mercadier, J.J. and Michel, J.B. The plasminogen-MMP system is more activated in the scar than in viable myocardium 3 months post-MI in the rat. J. Mol. Cell. Cardiol. 38 (2005) 193–204.

    Article  CAS  PubMed  Google Scholar 

  31. Baker, A.H., Edwards, D.R. and Murphy, G. Metalloproteinase inhibitors: biological actions and therapeutic opportunities. J. Cell. Sci. 115 (2002) 3719–3727.

    Article  CAS  PubMed  Google Scholar 

  32. Cheung C., Luo H., Yanagawa B., Leong H.S., Samarasekera D., Lai J.C., Suarez, A., Zhang, J. and McManus, B.M. Matrix metalloproteinases and tissue inhibitors of metalloproteinases in coxsackievirus-induced myocarditis. Cardiovasc. Pathol. 15 (2006) 63–74.

    Article  CAS  PubMed  Google Scholar 

  33. Li, J., Schwimmbeck, P.L., Tschope, C., Leschka, S., Husmann, L., Rutschow, S., Reichenbach, F., Noutsias, M., Kobalz, U., Poller, W., Spillmann, F., Zeichhardt, H., Schultheiss, H.P. and Pauschinger, M. Collagen degradation in a murine myocarditis model: relevance of matrix metalloproteinase in association with inflammatory induction. Cardiovasc. Res. 56 (2002) 235–247.

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations


Corresponding author

Correspondence to Qizhu Tang.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Tang, Q., Huang, J., Qian, H. et al. Microarray analysis reveals the role of matrix metalloproteinases in mouse experimental autoimmune myocarditis induced by cardiac myosin peptides. Cell Mol Biol Lett 12, 176–191 (2007).

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI:

Key words