OBJECTIVES: This study sought to develop a rapid method for the detection of activating autoantibodies directed against the beta1-adrenoceptor (anti-beta1-Abs) in patients with heart failure. BACKGROUND: The anti-beta1-Abs are supposed to play a pathophysiological role in heart failure. However, there is no reliable method for their detection. With a complex screening strategy (enzyme-linked immunosorbent assay, immunofluorescence, cyclic adenosine monophosphate [cAMP]-radioimmunoassay) we have previously identified antibodies targeting the second extracellular beta1-receptor loop (anti-beta1-EC(II)) in 13% of patients with ischemic cardiomyopathy (ICM) and in 26% with dilated cardiomyopathy (DCM). METHODS: To detect anti-beta1-Abs, we measured beta1-receptor-mediated increases in intracellular cAMP by fluorescence resonance energy transfer using a highly sensitive cAMP sensor (Epac1-based fluorescent cAMP sensor). RESULTS: The immunoglobulin G (IgG) prepared from 77 previously antibody-typed patients (22 ICM/55 DCM) and 50 matched control patients was analyzed. The IgG from all 22 previously anti-beta1-EC(II)-positive patients (5 ICM/17 DCM) induced a marked cAMP increase, indicating receptor activation (49.8 +/- 4.2% of maximal isoproterenol-induced signal). The IgG from control patients and 32 previously anti-beta1-EC(II)-negative patients (17 ICM/15 DCM) did not significantly affect cAMP. Surprisingly, our technology detected anti-beta1-Abs in 23 DCM patients formerly judged antibody-negative, but their cAMP signals were generally lower (31.3 +/- 6.8%) than in the previous group. "Low"-activator anti-beta1-Abs were blocked preferentially by peptides corresponding to the first, and "high"-activator anti-beta1-Abs by peptides corresponding to the second beta1-extracellular loop. Beta-blockers alone failed to fully prevent anti-beta1-EC(II)-induced receptor activation, which could be achieved, however, by the addition of beta1-EC(II) peptides. CONCLUSIONS: Our novel method of detecting anti-beta1-Abs proved to be fast and highly sensitive. It also revealed an insufficient ability of beta-blockers to prevent anti-beta1-EC(II)-induced receptor activation, which opens new venues for the research on anti-beta1-Abs and eventual treatment options in heart failure.
OBJECTIVES: This study sought to develop a rapid method for the detection of activating autoantibodies directed against the beta1-adrenoceptor (anti-beta1-Abs) in patients with heart failure. BACKGROUND: The anti-beta1-Abs are supposed to play a pathophysiological role in heart failure. However, there is no reliable method for their detection. With a complex screening strategy (enzyme-linked immunosorbent assay, immunofluorescence, cyclic adenosine monophosphate [cAMP]-radioimmunoassay) we have previously identified antibodies targeting the second extracellular beta1-receptor loop (anti-beta1-EC(II)) in 13% of patients with ischemic cardiomyopathy (ICM) and in 26% with dilated cardiomyopathy (DCM). METHODS: To detect anti-beta1-Abs, we measured beta1-receptor-mediated increases in intracellular cAMP by fluorescence resonance energy transfer using a highly sensitive cAMP sensor (Epac1-based fluorescent cAMP sensor). RESULTS: The immunoglobulin G (IgG) prepared from 77 previously antibody-typed patients (22 ICM/55 DCM) and 50 matched control patients was analyzed. The IgG from all 22 previously anti-beta1-EC(II)-positive patients (5 ICM/17 DCM) induced a marked cAMP increase, indicating receptor activation (49.8 +/- 4.2% of maximal isoproterenol-induced signal). The IgG from control patients and 32 previously anti-beta1-EC(II)-negative patients (17 ICM/15 DCM) did not significantly affect cAMP. Surprisingly, our technology detected anti-beta1-Abs in 23 DCM patients formerly judged antibody-negative, but their cAMP signals were generally lower (31.3 +/- 6.8%) than in the previous group. "Low"-activator anti-beta1-Abs were blocked preferentially by peptides corresponding to the first, and "high"-activator anti-beta1-Abs by peptides corresponding to the second beta1-extracellular loop. Beta-blockers alone failed to fully prevent anti-beta1-EC(II)-induced receptor activation, which could be achieved, however, by the addition of beta1-EC(II) peptides. CONCLUSIONS: Our novel method of detecting anti-beta1-Abs proved to be fast and highly sensitive. It also revealed an insufficient ability of beta-blockers to prevent anti-beta1-EC(II)-induced receptor activation, which opens new venues for the research on anti-beta1-Abs and eventual treatment options in heart failure.
Authors: Iris I Müller; Karin Klingel; Viacheslav O Nikolaev; R Jahns; Meinrad P Gawaz; Hans-Jörg Weig Journal: Clin Res Cardiol Date: 2008-07-16 Impact factor: 5.460
Authors: Hongliang Li; David C Kem; Sean Reim; Muneer Khan; Megan Vanderlinde-Wood; Caitlin Zillner; Daniel Collier; Campbell Liles; Michael A Hill; Madeleine W Cunningham; Christopher E Aston; Xichun Yu Journal: Hypertension Date: 2012-01-03 Impact factor: 10.190
Authors: Stavros Stavrakis; David C Kem; Eugene Patterson; Pedro Lozano; Shijun Huang; Bela Szabo; Madeleine W Cunningham; Ralph Lazzara; Xichun Yu Journal: Int J Cardiol Date: 2010-01-06 Impact factor: 4.164