Profiling humoral autoimmune repertoire of dilated cardiomyopathy (DCM) patients and development of a disease‐associated protein chip

Dilated cardiomyopathy (DCM) is a myocardial disease characterized by progressive depression of myocardial contractile function and ventricular dilatation. Thirty percent of DCM patients belong to the inherited genetic form; the rest may be idiopathic, viral, autoimmune, or immune‐mediated associated with a viral infection. Disturbances in humoral and cellular immunity have been described in cases of myocarditis and DCM. A number of autoantibodies against cardiac cell proteins have been identified in DCM. In this study, we have profiled the autoantibody repertoire of plasma from DCM patients against a human protein array consisting of 37 200 redundant, recombinant human proteins and performed qualitative and quantitative validation of these putative autoantigens on protein microarrays to identify novel putative DCM specific autoantigens. In addition to analyzing the whole IgG autoantibody repertoire, we have also analyzed the IgG3 antibody repertoire in the plasma samples to study the characteristics of IgG3 subclass antibodies. By combining screening of a protein expression library with protein microarray technology, we have detected 26 proteins identified by the IgG antibody repertoire and 6 proteins bound by the IgG3 subclass. Several of these autoantibodies found in plasma of DCM patients, such as the autoantibody against the Kv channel‐interacting protein, are associated with heart failure.

[1]  Eytan Domany,et al.  Functional immunomics: Microarray analysis of IgG autoantibody repertoires predicts the future response of mice to induced diabetes , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[2]  P. Richardson.  Assessment of myocardial damage in dilated cardiomyopathy. , 1996, European heart journal.

[3]  T. Ternynck,et al.  Characterization of autoantibody activities in sera anti‐DNA antibody and circulating immune complexes from 12 systemic lupus erythematosus patients , 1996, Journal of clinical laboratory analysis.

[4]  M. Yacoub,et al.  The expression of heat shock protein 60 in patients with dilated cardiomyophathy , 1999, Basic Research in Cardiology.

[5]  M. Yacoub,et al.  Subclass specificity of autoantibodies against myosin in patients with idiopathic dilated cardiomyopathy: pro-inflammatory antibodies in DCM patients. , 1999, Biochemical and biophysical research communications.

[6]  S. Olsson,et al.  The dispersion of repolarization in patients with ventricular tachycardia. A study using simultaneous monophasic action potential recordings from two sites in the right ventricle. , 1995, European heart journal.

[7]  T. Hedner,et al.  Autoantibodies against cardiac G-protein-coupled receptors define different populations with cardiomyopathies but not with hypertension. , 1994, Clinical immunology and immunopathology.

[8]  Joshua Labaer,et al.  The FLEXGene repository: exploiting the fruits of the genome projects by creating a needed resource to face the challenges of the post-genomic era. , 2002, Archives of medical research.

[9]  Marc Vidal,et al.  A cDNA encoding a pRB-binding protein with properties of the transcription factor E2F , 1992, Cell.

[10]  N. Dhalla,et al.  Alterations of sarcolemmal phospholipase D and phosphatidate phosphohydrolase in congestive heart failure. , 2002, Biochimica et biophysica acta.

[11]  T. Honjo,et al.  PD-1: an inhibitory immunoreceptor involved in peripheral tolerance. , 2001, Trends in immunology.

[12]  Lawrence Steinman,et al.  Proteomics technologies for the study of autoimmune disease. , 2002, Arthritis and rheumatism.

[13]  X. Qu,et al.  Characterization and tissue expression of a novel human gene npdc1. , 2001, Gene.

[14]  G. Wallukat,et al.  Immunohistological Changes in Dilated Cardiomyopathy Induced by Immunoadsorption Therapy and Subsequent Immunoglobulin Substitution , 2001, Circulation.

[15]  J. Omens,et al.  Protection against myocardial dysfunction after a brief ischemic period in transgenic mice expressing inducible heat shock protein 70. , 1998, The Journal of clinical investigation.

[16]  T. Komurasaki,et al.  A novel homocysteine-responsive gene, smap8, modulates mitogenesis in rat vascular smooth muscle cells. , 2003, European journal of biochemistry.

[17]  Dolores J Cahill,et al.  The impact of protein biochips and microarrays on the drug development process. , 2002, Drug discovery today.

[18]  D. Yellon,et al.  Overexpression of the rat inducible 70-kD heat stress protein in a transgenic mouse increases the resistance of the heart to ischemic injury. , 1995, The Journal of clinical investigation.

[19]  F. Bernier-Valentin,et al.  Evidence for anti-tubulin autoantibodies in the form of immune complexes in human sera. , 1988, Clinical and Experimental Immunology.

[20]  J. Mason,et al.  Advances in the understanding of myocarditis. , 2001, Circulation.

[21]  G. Wallukat,et al.  Potential Role of Autoantibodies Belonging to the Immunoglobulin G-3 Subclass in Cardiac Dysfunction Among Patients With Dilated Cardiomyopathy , 2002, Circulation.

[22]  I. Cohen,et al.  The natural autoantibody repertoire and autoimmune disease. , 2004, Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie.

[23]  M. Peakman,et al.  T Cell Activation by Coxsackievirus B4 Antigens in Type 1 Diabetes Mellitus: Evidence for Selective TCR Vβ Usage Without Superantigenic Activity1 , 2001, The Journal of Immunology.

[24]  David J. Moliterno,et al.  Evidence for substantial effect modification by gender in a large-scale genetic association study of the metabolic syndrome among coronary heart disease patients , 2003, Human Genetics.

[25]  R. Hullin,et al.  Regional expression and functional characterization of the L-type Ca2+-channel in myocardium from patients with end-stage heart failure and in non-failing human hearts. , 1999, Journal of molecular and cellular cardiology.

[26]  E. Dupont,et al.  NPDC-1, a regulator of neural cell proliferation and differentiation, interacts with E2F-1, reduces its binding to DNA and modulates its transcriptional activity , 2000, Oncogene.

[27]  W. Mckenna,et al.  Prognosis of idiopathic dilated cardiomyopathy. , 1995, Journal of cardiac failure.

[28]  S. Felix,et al.  Improvement of Cardiac Function after Immunoadsorption in Patients with Dilated Cardiomyopathy , 2001, Autoimmunity.

[29]  M. Morad,et al.  Antibody-mediated enhancement of calcium permeability in cardiac myocytes , 1988, The Journal of experimental medicine.

[30]  T. Miyata,et al.  Characterization of the human NDRG gene family: a newly identified member, NDRG4, is specifically expressed in brain and heart. , 2001, Genomics.

[31]  H. Lehrach,et al.  A Nonredundant Human Protein Chip for Antibody Screening and Serum Profiling* , 2003, Molecular & Cellular Proteomics.

[32]  P. Lazarow.  Peroxisome structure, function, and biogenesis--human patients and yeast mutants show strikingly similar defects in peroxisome biogenesis. , 1995, Journal of neuropathology and experimental neurology.

[33]  A. Coutinho,et al.  Self-reactive antibodies (natural autoantibodies) in healthy individuals. , 1998, Journal of immunological methods.

[34]  J. Goldman,et al.  Organ-specific cardiac autoantibodies in dilated cardiomyopathy--an update. , 1995, European heart journal.

[35]  S. Kaveri,et al.  Natural Antibodies to Factor VIII , 2000, Seminars in thrombosis and hemostasis.

[36]  Lawrence Steinman,et al.  Protein microarrays guide tolerizing DNA vaccine treatment of autoimmune encephalomyelitis , 2003, Nature Biotechnology.

[37]  S. Felix.  Immunoadsorption in dilated cardiomyopathy. , 2000, Ernst Schering Research Foundation workshop.

[38]  K. Chien,et al.  Genetic modifiers of cardiac arrhythmias. , 2003, Trends in molecular medicine.

[39]  R. Mestril,et al.  Overexpression of heat shock protein 72 in transgenic mice decreases infarct size in vivo. , 1996, Circulation.

[40]  M. Pucéat,et al.  Signalling by protein kinase C isoforms in the heart , 1996, Molecular and Cellular Biochemistry.

[41]  A. Gramolini,et al.  Cardiac-specific overexpression of sarcolipin inhibits sarco(endo)plasmic reticulum Ca2+ ATPase (SERCA2a) activity and impairs cardiac function in mice. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[42]  Yoshimasa Tanaka,et al.  Autoantibodies against cardiac troponin I are responsible for dilated cardiomyopathy in PD-1-deficient mice , 2003, Nature Medicine.

[43]  J. Ross,et al.  A Defect in the Kv Channel-Interacting Protein 2 (KChIP2) Gene Leads to a Complete Loss of I to and Confers Susceptibility to Ventricular Tachycardia , 2001, Cell.

[44]  H. Schultheiss,et al.  Antibodies to ADP-ATP carrier--an autoantigen in myocarditis and dilated cardiomyopathy--impair cardiac function. , 1990, Circulation.

[45]  V. Álvarez,et al.  Variation in the lipoprotein receptor-related protein, α2-macroglobulin and lipoprotein receptor-associatedprotein genes in relation to plasma lipid levels and riskof early myocardial infarction , 2002, Coronary artery disease.

[46]  Martin J. Lohse,et al.  Direct evidence for a β1-adrenergic receptor–directed autoimmune attack as a cause of idiopathic dilated cardiomyopathy , 2004 .

[47]  U. Utz,et al.  Treatment of experimental encephalomyelitis with a peptide analogue of myelin basic protein , 1996, Nature.

[48]  Thomas L. Madden,et al.  Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. , 1997, Nucleic acids research.

[49]  H. Lehrach,et al.  A human cDNA library for high-throughput protein expression screening. , 2000, Genomics.

[50]  G. Wallukat,et al.  Removal of cardiodepressant antibodies in dilated cardiomyopathy by immunoadsorption. , 2002, Journal of the American College of Cardiology.

[51]  T. Waldmann,et al.  Distinct pathways involving the FK506-binding proteins 12 and 12.6 underlie IL-2-versus IL-15-mediated proliferation of T cells , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[52]  M. Fu,et al.  Is cardiomyopathy an autoimmune disease? , 2002, The Keio journal of medicine.

[53]  H. Lehrach,et al.  A method for global protein expression and antibody screening on high-density filters of an arrayed cDNA library. , 1998, Nucleic acids research.

[54]  P. Brown,et al.  Autoantigen microarrays for multiplex characterization of autoantibody responses , 2002, Nature Medicine.

[55]  I. Goldenberg,et al.  Autoantibodies against beta-adrenoceptors in human idiopathic dilated cardiomyopathy. , 1989, Circulation research.

[56]  Y. Liao.  Functional analysis of autoantibodies against ADP/ATP carrier from dilated cardiomyopathy. , 1996, International journal of cardiology.

[57]  H. Lehrach,et al.  Generation of minimal protein identifiers of proteins from two‐dimensional gels and recombinant proteins , 2002, Electrophoresis.

[58]  D. Cahill,et al.  Protein arrays and their role in proteomics. , 2003, Advances in biochemical engineering/biotechnology.

[59]  E. Nogales,et al.  Tubulin and FtsZ form a distinct family of GTPases , 1998, Nature Structural Biology.

[60]  H. Lehrach,et al.  Protein microarrays for gene expression and antibody screening. , 1999, Analytical biochemistry.

[61]  E. Bollano,et al.  Induction of Cardiomyopathy in Severe Combined Immunodeficiency Mice by Transfer of Lymphocytes from Patients with Idiopathic Dilated Cardiomyopathy , 2000 .

[62]  Stuart L. Schreiber,et al.  Calcineurin is a common target of cyclophilin-cyclosporin A and FKBP-FK506 complexes , 1991, Cell.

[63]  Yanhui Hu,et al.  Proteome-scale purification of human proteins from bacteria , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[64]  R. Hetzer,et al.  Immunoglobulin adsorption in patients with idiopathic dilated cardiomyopathy. , 2000, Circulation.

[65]  E. Tan,et al.  Autoantibodies in the diagnosis of systemicrheumatic diseases , 1995 .

[66]  W. Mckenna,et al.  Identification of alpha- and beta-cardiac myosin heavy chain isoforms as major autoantigens in dilated cardiomyopathy. , 1992, Circulation.

[67]  S. Kaveri,et al.  Autoantibodies to heat shock protein 90 in the human natural antibody repertoire. , 2002, International immunology.

[68]  E. Saggerson,et al.  Glycerolipid metabolizing enzymes in rat ventricle and in cardiac myocytes. , 1997, Biochimica et biophysica acta.