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Interleukin-6 receptor inhibition modulates the immune reaction and restores titin phosphorylation in experimental myocarditis

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Abstract

Increased levels of interleukin-6 (IL-6) have been observed in patients with acute myocarditis and are associated with poor prognosis. This study was designed to examine whether treatment with anti-IL-6 receptor antibody improves cardiac dysfunction and left ventricular (LV) remodeling in experimental Coxsackie virus B3 (CVB3)-induced myocarditis. C57BL6/J mice were subjected to acute CVB3 infection. One day after viral infection mice were treated with a single injection of an anti-IL-6 receptor antibody (MR16-1, tocilizumab) or control IgG. Seven days after viral infection, LV function was examined by conductance catheter technique, cardiac remodeling assessed by estimation of titin phosphorylation, cardiac fibrosis, and inflammatory and antiviral response by immunohistochemistry, RT-PCR and cell culture experiments. Compared to controls, infected mice displayed an impaired systolic and diastolic LV function associated with an increase in cardiac inflammation, fibrosis and impaired titin phosphorylation. IL-6 receptor blockade led to a shift of the immune response to a Th1 direction and significant reduction of viral load. In addition, cardiac immune response, extracellular matrix regulation and titin function improved, resulting in a preserved LV function. IL-6 receptor blockade exerts cardiac beneficial effects by antiviral and immunomodulatory actions after induction of an acute murine CVB3 virus myocarditis.

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References

  1. Bishu K, Hamdani N, Mohammed SF, Kruger M, Ohtani T, Ogut O, Brozovich FV, Burnett JC Jr, Linke WA, Redfield MM (2011) Sildenafil and B-type natriuretic peptide acutely phosphorylate titin and improve diastolic distensibility in vivo. Circulation 124:2882–2891. doi:10.1161/CIRCULATIONAHA.111.048520

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  2. Boengler K, Hilfiker-Kleiner D, Drexler H, Heusch G, Schulz R (2008) The myocardial JAK/STAT pathway: from protection to failure. Pharmacol Ther 120:172–185. doi:10.1016/j.pharmthera.2008.08.002

    Article  CAS  PubMed  Google Scholar 

  3. Chae CU, Lee RT, Rifai N, Ridker PM (2001) Blood pressure and inflammation in apparently healthy men. Hypertension 38:399–403. doi:10.1161/01.HYP.38.3.399

    Article  CAS  PubMed  Google Scholar 

  4. Choy E (2009) RADIATE: more treatment options for patients with an inadequate response to tumor necrosis factor antagonists. Nat Clin Pract Rheumatol 5:66–67. doi:10.1038/ncprheum0984

    Article  CAS  PubMed  Google Scholar 

  5. Esfandiarei M, McManus BM (2008) Molecular biology and pathogenesis of viral myocarditis. Annu Rev Pathol 3:127–155. doi:10.1146/annurev.pathmechdis.3.121806.151534

    Article  CAS  PubMed  Google Scholar 

  6. Fairweather D, Rose NR (2005) Inflammatory heart disease: a role for cytokines. Lupus 14:646–651. doi:10.1191/0961203305lu2192oa

    Article  CAS  PubMed  Google Scholar 

  7. Fairweather D, Stafford KA, Sung YK (2012) Update on coxsackievirus B3 myocarditis. Curr Opin Rheumatol 24:401–407. doi:10.1097/BOR.0b013e328353372d

    Article  PubMed  Google Scholar 

  8. Febbraio MA, Hiscock N, Sacchetti M, Fischer CP, Pedersen BK (2004) Interleukin-6 is a novel factor mediating glucose homeostasis during skeletal muscle contraction. Diabetes 53:1643–1648. doi:10.2337/diabetes.53.7.1643

    Article  CAS  PubMed  Google Scholar 

  9. Fischer P, Hilfiker-Kleiner D (2007) Survival pathways in hypertrophy and heart failure: the gp130-STAT axis. Basic Res Cardiol 102:393–411. doi:10.1007/s00395-007-0658-z

    Article  CAS  PubMed  Google Scholar 

  10. Gallucci RM, Lee EG, Tomasek JJ (2006) IL-6 modulates alpha-smooth muscle actin expression in dermal fibroblasts from IL-6-deficient mice. J Invest Dermatol 126:561–568. doi:10.1038/sj.jid.5700109

    Article  CAS  PubMed  Google Scholar 

  11. Heinrich PC, Behrmann I, Haan S, Hermanns HM, Muller-Newen G, Schaper F (2003) Principles of interleukin (IL)-6-type cytokine signalling and its regulation. Biochem J 374:1–20. doi:10.1042/BJ20030407

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  12. Heymans S (2006) Inflammation and cardiac remodeling during viral myocarditis. Ernst Schering Res Found Workshop, pp 197–218

  13. Heymans S, Hirsch E, Anker SD, Aukrust P, Balligand JL, Cohen-Tervaert JW, Drexler H, Filippatos G, Felix SB, Gullestad L, Hilfiker-Kleiner D, Janssens S, Latini R, Neubauer G, Paulus WJ, Pieske B, Ponikowski P, Schroen B, Schultheiss HP, Tschope C, Van Bilsen M, Zannad F, McMurray J, Shah AM (2009) Inflammation as a therapeutic target in heart failure? A scientific statement from the Translational Research Committee of the Heart Failure Association of the European Society of Cardiology. Eur J Heart Fail 11:119–129. doi:10.1093/eurjhf/hfn043

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  14. Hirota H, Yoshida K, Kishimoto T, Taga T (1995) Continuous activation of gp130, a signal-transducing receptor component for interleukin 6-related cytokines, causes myocardial hypertrophy in mice. Proc Natl Acad Sci USA 92:4862–4866

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  15. Jaber WA, Maniu C, Krysiak J, Shapiro BP, Meyer DM, Linke WA, Redfield MM (2008) Titin isoforms, extracellular matrix, and global chamber remodeling in experimental dilated cardiomyopathy: functional implications and mechanistic insight. Circ Heart Fail 1:192–199. doi:10.1161/CIRCHEARTFAILURE.108.768465

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  16. Jones SA, Scheller J, Rose-John S (2011) Therapeutic strategies for the clinical blockade of IL-6/gp130 signaling. J Clin Invest 121:3375–3383. doi:10.1172/JCI57158

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  17. Kanda T, Takahashi T (2004) Interleukin-6 and cardiovascular diseases. Jpn Heart J 45:183–193. doi:10.1536/jhj.45.183

    Article  CAS  PubMed  Google Scholar 

  18. Katsume A, Saito H, Yamada Y, Yorozu K, Ueda O, Akamatsu K, Nishimoto N, Kishimoto T, Yoshizaki K, Ohsugi Y (2002) Anti-interleukin 6 (IL-6) receptor antibody suppresses Castleman’s disease like symptoms emerged in IL-6 transgenic mice. Cytokine 20:304–311. doi:10.1006/cyto.2002.2012

    Article  CAS  PubMed  Google Scholar 

  19. Kleinbongard P, Heusch G, Schulz R (2010) TNFalpha in atherosclerosis, myocardial ischemia/reperfusion and heart failure. Pharmacol Ther 127:295–314. doi:10.1016/j.pharmthera.2010.05.002

    Article  CAS  PubMed  Google Scholar 

  20. Klingel K, Hohenadl C, Canu A, Albrecht M, Seemann M, Mall G, Kandolf R (1992) Ongoing enterovirus-induced myocarditis is associated with persistent heart muscle infection: quantitative analysis of virus replication, tissue damage, and inflammation. Proc Natl Acad Sci USA 89:314–318

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  21. Klover PJ, Clementi AH, Mooney RA (2005) Interleukin-6 depletion selectively improves hepatic insulin action in obesity. Endocrinology 146:3417–3427. doi:10.1210/en.2004-1468

    Article  CAS  PubMed  Google Scholar 

  22. Kruger M, Linke WA (2009) Titin-based mechanical signalling in normal and failing myocardium. J Mol Cell Cardiol 46:490–498. doi:10.1016/j.yjmcc.2009.01.004

    Article  PubMed  Google Scholar 

  23. Lee DL, Sturgis LC, Labazi H, Osborne JB Jr, Fleming C, Pollock JS, Manhiani M, Imig JD, Brands MW (2006) Angiotensin II hypertension is attenuated in interleukin-6 knockout mice. Am J Physiol Heart Circ Physiol 290:H935–H940. doi:10.1152/ajpheart.00708.2005

    Article  CAS  PubMed  Google Scholar 

  24. Linke WA, Hamdani N (2014) Gigantic business: titin properties and function through thick and thin. Circ Res 114:1052–1068. doi:10.1161/CIRCRESAHA.114.301286

    Article  CAS  PubMed  Google Scholar 

  25. Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods 25:402–408. doi:10.1006/meth.2001.1262

    Article  CAS  PubMed  Google Scholar 

  26. McKenzie BS, Kastelein RA, Cua DJ (2006) Understanding the IL-23-IL-17 immune pathway. Trends Immunol 27:17–23. doi:10.1016/j.it.2005.10.003

    Article  CAS  PubMed  Google Scholar 

  27. Nagahama KY, Togo S, Holz O, Magnussen H, Liu X, Seyama K, Takahashi K, Rennard SI (2013) Oncostatin M modulates fibroblast function via signal transducers and activators of transcription proteins-3. Am J Respir Cell Mol Biol 49:582–591. doi:10.1165/rcmb.2012-0460OC

    Article  CAS  PubMed  Google Scholar 

  28. Nowell MA, Richards PJ, Fielding CA, Ognjanovic S, Topley N, Williams AS, Bryant-Greenwood G, Jones SA (2006) Regulation of pre-B cell colony-enhancing factor by STAT-3-dependent interleukin-6 trans-signaling: implications in the pathogenesis of rheumatoid arthritis. Arthritis Rheum 54:2084–2095. doi:10.1002/art.21942

    Article  CAS  PubMed  Google Scholar 

  29. Okazaki M, Yamada Y, Nishimoto N, Yoshizaki K, Mihara M (2002) Characterization of anti-mouse interleukin-6 receptor antibody. Immunol Lett 84:231–240. doi:10.1016/S0165-2478(02)00202-X

    Article  CAS  PubMed  Google Scholar 

  30. Pal M, Febbraio MA, Whitham M (2014) From cytokine to myokine: the emerging role of interleukin-6 in metabolic regulation. Immunol Cell Biol 92:331–339. doi:10.1038/icb.2014.16

    Article  CAS  PubMed  Google Scholar 

  31. Poffenberger MC, Horwitz MS (2009) IL-6 during viral-induced chronic autoimmune myocarditis. Ann NY Acad Sci 1173:318–325. doi:10.1111/j.1749-6632.2009.04850.x

    Article  CAS  PubMed  Google Scholar 

  32. Roll P, Muhammad K, Schumann M, Kleinert S, Einsele H, Dorner T, Tony HP (2011) In vivo effects of the anti-interleukin-6 receptor inhibitor tocilizumab on the B cell compartment. Arthritis Rheum 63:1255–1264. doi:10.1002/art.30242

    Article  CAS  PubMed  Google Scholar 

  33. Rosamond W, Flegal K, Furie K, Go A, Greenlund K, Haase N, Hailpern SM, Ho M, Howard V, Kissela B, Kittner S, Lloyd-Jones D, McDermott M, Meigs J, Moy C, Nichol G, O’Donnell C, Roger V, Sorlie P, Steinberger J, Thom T, Wilson M, Hong Y (2008) Heart disease and stroke statistics—2008 update: a report from the American Heart Association Statistics Committee and Stroke Statistics Subcommittee. Circulation 117:e25–e146. doi:10.1161/CIRCULATIONAHA.107.187998

    Article  PubMed  Google Scholar 

  34. Sarkar S, Vellaichamy E, Young D, Sen S (2004) Influence of cytokines and growth factors in ANG II-mediated collagen upregulation by fibroblasts in rats: role of myocytes. Am J Physiol Heart Circ Physiol 287:H107–H117. doi:10.1152/ajpheart.00763.2003

    Article  CAS  PubMed  Google Scholar 

  35. Savvatis K, Pappritz K, Becher PM, Lindner D, Zietsch C, Volk HD, Westermann D, Schultheiss HP, Tschope C (2014) Interleukin-23 deficiency leads to impaired wound healing and adverse prognosis after myocardial infarction. Circ Heart Fail 7:161–171. doi:10.1161/CIRCHEARTFAILURE.113.000604

    Article  CAS  PubMed  Google Scholar 

  36. Savvatis K, van Linthout S, Miteva K, Pappritz K, Westermann D, Schefold JC, Fusch G, Weithauser A, Rauch U, Becher PM, Klingel K, Ringe J, Kurtz A, Schultheiss HP, Tschope C (2012) Mesenchymal stromal cells but not cardiac fibroblasts exert beneficial systemic immunomodulatory effects in experimental myocarditis. PLoS ONE 7:e41047. doi:10.1371/journal.pone.0041047

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  37. Siwik DA, Chang DL, Colucci WS (2000) Interleukin-1beta and tumor necrosis factor-alpha decrease collagen synthesis and increase matrix metalloproteinase activity in cardiac fibroblasts in vitro. Circ Res 86:1259–1265. doi:10.1161/01.RES.86.12.1259

    Article  CAS  PubMed  Google Scholar 

  38. Tanaka T, Kanda T, McManus BM, Kanai H, Akiyama H, Sekiguchi K, Yokoyama T, Kurabayashi M (2001) Overexpression of interleukin-6 aggravates viral myocarditis: impaired increase in tumor necrosis factor-alpha. J Mol Cell Cardiol 33:1627–1635. doi:10.1006/jmcc.2001.1428

    Article  CAS  PubMed  Google Scholar 

  39. Tsutamoto T, Hisanaga T, Wada A, Maeda K, Ohnishi M, Fukai D, Mabuchi N, Sawaki M, Kinoshita M (1998) Interleukin-6 spillover in the peripheral circulation increases with the severity of heart failure, and the high plasma level of interleukin-6 is an important prognostic predictor in patients with congestive heart failure. J Am Coll Cardiol 31:391–398. doi:10.1016/S0735-1097(97)00494-4

    Article  CAS  PubMed  Google Scholar 

  40. van Nieuwenhoven FA, Turner NA (2013) The role of cardiac fibroblasts in the transition from inflammation to fibrosis following myocardial infarction. Vascul Pharmacol 58:182–188. doi:10.1016/j.vph.2012.07.003

    Article  PubMed  Google Scholar 

  41. Veldhoen M, Hocking RJ, Atkins CJ, Locksley RM, Stockinger B (2006) TGFbeta in the context of an inflammatory cytokine milieu supports de novo differentiation of IL-17-producing T cells. Immunity 24:179–189. doi:10.1016/j.immuni.2006.01.001

    Article  CAS  PubMed  Google Scholar 

  42. Westermann D, Lindner D, Kasner M, Zietsch C, Savvatis K, Escher F, von Schlippenbach J, Skurk C, Steendijk P, Riad A, Poller W, Schultheiss HP, Tschope C (2011) Cardiac inflammation contributes to changes in the extracellular matrix in patients with heart failure and normal ejection fraction. Circ Heart Fail 4:44–52. doi:10.1161/CIRCHEARTFAILURE.109.931451

    Article  PubMed  Google Scholar 

  43. Westermann D, Savvatis K, Schultheiss HP, Tschope C (2010) Immunomodulation and matrix metalloproteinases in viral myocarditis. J Mol Cell Cardiol 48:468–473. doi:10.1016/j.yjmcc.2009.08.019

    Article  CAS  PubMed  Google Scholar 

  44. Westermann D, Walther T, Savvatis K, Escher F, Sobirey M, Riad A, Bader M, Schultheiss HP, Tschope C (2009) Gene deletion of the kinin receptor B1 attenuates cardiac inflammation and fibrosis during the development of experimental diabetic cardiomyopathy. Diabetes 58:1373–1381. doi:10.2337/db08-0329

    Article  PubMed Central  PubMed  Google Scholar 

  45. Woodruff JF (1980) Viral myocarditis. A review. Am J Pathol 101:425–484

    CAS  PubMed Central  PubMed  Google Scholar 

  46. Yajima T, Knowlton KU (2009) Viral myocarditis: from the perspective of the virus. Circulation 119:2615–2624. doi:10.1161/CIRCULATIONAHA.108.766022

    Article  PubMed  Google Scholar 

  47. Yang F, Wu WF, Yan YL, Pang Y, Kong Q, Huang YL (2011) Expression of IL-23/Th17 pathway in a murine model of Coxsackie virus B3-induced viral myocarditis. Virol J 8:301. doi:10.1186/1743-422X-8-301

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  48. Yuan J, Yu M, Lin QW, Cao AL, Yu X, Dong JH, Wang JP, Zhang JH, Wang M, Guo HP, Cheng X, Liao YH (2010) Th17 cells contribute to viral replication in coxsackievirus B3-induced acute viral myocarditis. J Immunol 185:4004–4010. doi:10.4049/jimmunol.1001718

    Article  CAS  PubMed  Google Scholar 

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Acknowledgments

We thank Kerstin Puhl and Georg Zingler for excellent technical assistance. The monoclonal anti-IL-6 receptor antibody (MR16-1) was a kind gift from Chugai Pharmaceutical Co. Ltd., Tokyo, Japan. This study was funded by the Deutsche Forschungsgesellschaft(SFB-TR-19: Z3, Z4), the “Berlin Center For Regenerative Therapies” (BCRT) and the “Deutsches Zentrum für Herzkreislaufforschung” (DZHK).

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None declared.

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Authors and Affiliations

  1. Department of Cardiology and Pneumology, Charité, Universitätsmedizin Berlin, Campus Benjamin Franklin, Berlin, Germany

    Konstantinos Savvatis, Irene Müller, Matthias Fröhlich, Kathleen Pappritz, Christin Zietsch, Heinz-Peter Schultheiss & Carsten Tschöpe

  2. Berlin-Brandenburg Center for Regenerative Therapies, Charité, University Medicine Berlin, Campus Virchow, Berlin, Germany

    Konstantinos Savvatis, Kathleen Pappritz, Sophie Van Linthout & Carsten Tschöpe

  3. Department of Cardiology and Angiology, Hannover Medical School, Hannover, Germany

    Karsten Grote

  4. Department of Internal Medicine and Cardiology, Philipps University, Marburg, Germany

    Bernhard Schieffer

  5. Department of Molecular Pathology, University Hospital Tuebingen, Tuebingen, Germany

    Karin Klingel

  6. Department of Cardiovascular Physiology, Institute of Physiology, Ruhr University, Bochum, Germany

    Nazha Hamdani & Wolfgang A. Linke

  7. DZHK, German Center for Cardiovascular Research, Partner Site Berlin, Charité, Berlin, Germany

    Irene Müller & Carsten Tschöpe

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  1. Konstantinos Savvatis
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  2. Irene Müller
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  3. Matthias Fröhlich
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Correspondence to Konstantinos Savvatis.

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Savvatis, K., Müller, I., Fröhlich, M. et al. Interleukin-6 receptor inhibition modulates the immune reaction and restores titin phosphorylation in experimental myocarditis. Basic Res Cardiol 109, 449 (2014). https://doi.org/10.1007/s00395-014-0449-2

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  • DOI: https://doi.org/10.1007/s00395-014-0449-2

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