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Removal of cardiodepressant antibodies in dilated cardiomyopathy by immunoadsorption

Stephan B. Felix, MD^*,*, Alexander Staudt, MD^*, Martin Landsberger, PhD^*, Yvonne Grosse, MD^*, Verena Stangl, MD, Thomas Spielhagen, MD, Gerd Wallukat, PhD, Klaus D. Wernecke, PhD, Gert Baumann, MD and Karl Stangl, MD

^* Klinik für Innere Medizin B, Universität Greifswald, Greifswald, Germany
Medizinische Klinik, Charité, Campus-Mitte, Berlin, Germany
Max Delbrück Zentrum für Molekulare Kardiologie, Berlin, Germany
Institut für Medizinische Biometrie, Charité, Humboldt-Universität, Berlin, Germany

View larger version (30K): [in a new window]   Figure 1 (A) Effects of the indicated dilutions of CE on isolated, field-stimulated rat cardiomyocytes. The CE was collected on day 1 during the first regeneration cycle of IA, session 1. Changes of Ca2+ transients (peak systolic rfu – diastolic rfu) (left plot) and systolic cell shortening (right plot) during superfusion of CE from healthy blood donors (control subjects = solid squares, n = 9) and of CE from patients with DCM (solid triangles, n = 11). Data (percent changes from baseline) are presented as the mean value ± SEM. *p < 0.05, **p < 0.01 and *** p < 0.001 vs. control subjects. (B) Correlation of effects of CE (collected on day 1, dilution 1:5) from patients with DCM on Ca2+ transients and systolic cell shortening of isolated, field-stimulated rat cardiomyocytes, with the increase in CI during session 1 of IA. CI = confidence interval.

View larger version (77K): [in a new window]   Figure 2 Equal amounts of immunoprecipitated myocardial proteins were loaded onto 5% to 20% sodium dodecyl sulfate–polyacrylamide gradient gels and visualized by silver staining. Immunoprecipitation was performed with sepharose-coupled antibodies collected during the first regeneration cycle of session 1 of IA from the CE of three patients with DCM and with sepharose-coupled antibodies obtained from the combined CE of two healthy blood donors. Immunoprecipitation with myocardial membrane-associated protein fraction (A) or with myocardial membrane protein fraction (B). M = molecular weight marker; AG = antigen (myocardial protein fraction) alone, no antibodies added; P1–3 = sepharose-coupled antibodies obtained from the CE of a patient with DCM plus myocardial protein fraction; C = sepharose-coupled antibodies obtained from the CE of control subjects (combined CE of two healthy male blood donors) plus myocardial protein fraction; AB = sepharose-coupled antibodies alone.