Enhanced expression and autoimmunity of recombination signal binding protein-jkappa in human dilated cardiomyopathy.
Dilated cardiomyopathy (DCM) is a major cause of heart failure in younger individuals. Its prognosis is poor with 40-50% of patients dying within 2 years after diagnosis. Although the etiology of DCM is poorly understood, there is increasing evidence that DCM may represent an autoimmune disease in a significant subset of patients. In order to identify candidate antigens in DCM, we applied a molecular strategy which combines recombinant expression cloning and autoimmunological screening procedures. A left ventricle from a male DCM patient was explanted at heart transplantation and a human DCM left ventricular cDNA-expression library was constructed. 2 x 10(6) clones were immunologically screened with serum collected from the same patient prior transplantation. Subsequent rounds of screening and purification allowed isolation of a positive clone which was sequenced and identified as Recombination Signal Binding Protein-jkappa (RBP-jkappa). RBP-jkappa is an already identified transcription factor, e.g., involved in Epstein-Barr-virus-induced immortalization processes. Radioactively labeled RBP-jkappa protein was synthesized via in vitro translation using the isolated RBP-jkappa cDNA. This RBP-jkappa protein was used for immunoprecipitation reactions to screen sera of healthy controls and patients suffering of DCM for the presence of RBP-jkappa autoantibodies. Analysis revealed that only 31% (n = 16) of healthy but 70.6% of DCM patients (n = 17) carry an autoantibody against RBP-jkappa. Patients suffering from ischemic cardiomyopathy showed a prevalence of 22% of RBP-jkappa autoantibodies. Western analysis with an monoclonal antibody raised against RBP-jkappa showed that RBP-jkappa was overexpressed to 488 +/- 140% in DCM hearts compared to non-failing controls (n = 8). Autologous immunological screening of a cDNA expression library is a powerful and novel technology to gain insights into the etiology of human idiopathic DCM. Human DCM displays an autoimmune response against RBP-jkappa and an overexpression of RBP-jkappa. Since RBP-jkappa is involved in cellular immortalization and exerts antiapoptotic effects, the increased RBP-jkappa autoantibody level during DCM may inhibit this growth-regulating feature of RBP-jkappa. In this setting, enhanced myocardial RBP-jkappa expression could represent a compensatory but ineffective response to counteract the increased rate of apoptosis in DCM. Furthermore, RBP-jkappa may be a useful diagnostic marker for DCM.
Nickenig
G
G
Klinik III für Innere Medizin, Institut für Virologie, Universität Köln, Joseph-Stelzmann-Strasse 9, Köln, 50924, Germany.
Wolff
M
M
Stäblein
A
A
Pfister
H
H
Böhm
M
M
D000368:Aged; D015551:Autoimmunity / Q000276:immunology*; D015153:Blotting, Western; D002311:Cardiomyopathy, Dilated / Q000276:immunology*; D003001:Cloning, Molecular; D004268:DNA-Binding Proteins / Q000097:blood / Q000235:genetics / Q000276:immunology*; D006352:Heart Ventricles / Q000378:metabolism; D006801:Humans; D051819:Immunoglobulin J Recombination Signal Sequence-Binding Protein; D008297:Male; D008875:Middle Aged; D009206:Myocardium / Q000276:immunology; D009687:Nuclear Proteins; D011233:Precipitin Tests; D014157:Transcription Factors
D004268:DNA-Binding Proteins; D051819:Immunoglobulin J Recombination Signal Sequence-Binding Protein; D009687:Nuclear Proteins; C497570:RBPJ protein, human; D014157:Transcription Factors