Startseite Medizin A standardised FACS assay based on native, receptor transfected cells for the clinical diagnosis and monitoring of β1-adrenergic receptor autoantibodies in human heart disease
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A standardised FACS assay based on native, receptor transfected cells for the clinical diagnosis and monitoring of β1-adrenergic receptor autoantibodies in human heart disease

  • Beatrice Bornholz , Thomas Benninghaus , Yvonne Reinke , Stephan B. Felix , Dirk Roggenbuck , Valérie Jahns-Boivin , Roland Jahns und Fritz Boege EMAIL logo
Veröffentlicht/Copyright: 25. September 2015
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Clinical Chemistry and Laboratory Medicine (CCLM)
Aus der Zeitschrift Clinical Chemistry and Laboratory Medicine (CCLM) Band 54 Heft 4

Abstract

Background: Autoantibodies against β1-adrenergic receptors (β1AR) that stimulate cardiac cAMP-production play a causal role in the pathogenesis of human heart failure. Patients can be subjected to specific therapies, if the presence of potentially cardio-noxious β1AR-autoantibodies is reliably diagnosed. This requires assessment of IgG-interactions with the native β1AR because β1AR-autoantibodies target a conformational epitope inadequately presented by denatured receptors or linear peptides. Here, we report on a standardised diagnostic procedure for the assessment of β1AR-autoantibodies in heart failure patients, which is based on IgG-binding to native human β1AR.

Methods: Good laboratory practice (GLP)-conform measurement of β1AR-autoantibodies was based on flow-cytometric quantification of differential IgG-binding to native HT1080 cells overexpressing biofluorescent human β1AR or not. Receptor-specific IgG-binding was derived from IgG-related median fluorescence of β1AR-positive cells corrected for background staining of β1AR-negative cells admixed to each measurement. The slope of IgG binding at two different concentrations was used as measure for the titre/avidity of β1AR-autoantibodies.

Results: Sensitivity and specificity of the novel procedure for high β1AR-autoantibody levels in dilated cardiomyopathy patients (n=40, NYHA class III-IV) relative to n=40 matched healthy subjects was >90%. It was similar to functional assays considered the gold standard and vastly superior to existing screening-procedures employing fixed cells or linear receptor-peptides as auto-antigenic targets. Inter-assay scatter was 7%–15% and linear dilution recovery was within ±10% of expected values throughout.

Conclusions: The novel assay possibly provides a tool to determine true prevalence and clinical impact of β1AR-autoantibodies. Furthermore, it may serve as companion diagnostic for therapies specifically directed at β1AR-autoantibodies.

Keywords: β1-adrenergic receptors; autoantibodies; companion assay; diagnostic testing; dilated cardiomyopathy; targeted therapy
Corresponding author: Fritz Boege, Institute for Clinical Chemistry and Laboratory Diagnostics, Medical Faculty, Heinrich Heine University, Moorenstr. 5, 40225 Düsseldorf, Germany, Phone: +49-211-8118290, Fax: +49-211-8118021, E-mail:

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Supplemental Material:

The online version of this article (DOI: 10.1515/cclm-2015-0603) offers supplementary material, available to authorized users.

Received: 2015-6-26
Accepted: 2015-8-31
Published Online: 2015-9-25
Published in Print: 2016-4-1

©2016 by De Gruyter

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https://doi.org/10.1515/cclm-2015-0603
Schlagwörter für diesen Artikel
β1-adrenergic receptors; autoantibodies; companion assay; diagnostic testing; dilated cardiomyopathy; targeted therapy

Artikel in diesem Heft

  1. Frontmatter
  2. Editorial
  3. Macroprolactin: searching for a needle in a haystack?
  4. Review
  5. Laboratory medicine in the new healthcare environment
  6. Opinion Paper
  7. Developing GRADE outcome-based recommendations about diagnostic tests: a key role in laboratory medicine policies
  8. EFLM Survey
  9. Accreditation process in European countries – an EFLM survey
  10. Genetics and Molecular Diagnostics
  11. HB Puerta del Sol [HBA1:c.148A>C], HB Valdecilla [HBA2:c.3G>T], HB Gran Vía [HBA2:c.98T>G], HB Macarena [HBA2:c.358C>T] and HB El Retiro [HBA2:c.364_366dupGTG]: description of five new hemoglobinopathies
  12. General Clinical Chemistry and Laboratory Medicine
  13. A candidate reference method using ICP-MS for sweat chloride quantification
  14. Quantification of hemoglobin A1c by off-line HPLC separation and liquid chromatography-tandem mass spectrometry: a modification of the IFCC reference measurement procedure
  15. Comparison between B·R·A·H·M·S PCT direct, a new sensitive point-of-care testing device for rapid quantification of procalcitonin in emergency department patients and established reference methods – a prospective multinational trial
  16. Evaluation of a hand-held blood gas analyzer for rapid determination of blood gases, electrolytes and metabolites in intensive care setting
  17. Use of a faecal immunochemical test for haemoglobin can aid in the investigation of patients with lower abdominal symptoms
  18. A possible cause of the variable detectability of macroprolactin by different immunoassay systems
  19. A multicenter comparison of whole blood vitamin B6 assays
  20. Analytical and clinical performance of the new Fujirebio 25-OH vitamin D assay, a comparison with liquid chromatography-tandem mass spectrometry (LC-MS/MS) and three other automated assays
  21. Development and validation of a 2nd tier test for identification of purine nucleoside phosphorylase deficiency patients during expanded newborn screening by liquid chromatography-tandem mass spectrometry
  22. A cross-sectional study of biomarkers of exposure and effect in smokers and moist snuff consumers
  23. Reference Values and Biological Variations
  24. Pediatric reference value distributions and covariate-stratified reference intervals for 29 endocrine and special chemistry biomarkers on the Beckman Coulter Immunoassay Systems: a CALIPER study of healthy community children
  25. Development of reference intervals for serum alkaline phosphatase among adults in Southern China traced to the new IFCC reference measurement procedure
  26. Birth season predicts the values of red blood cell distribution width (RDW) in adulthood
  27. Cardiovascular Diseases
  28. Cardiovascular risk stratification in hemodialysis patients in the era of highly sensitive troponins: should we choose between hs-troponin I and hs-troponin T?
  29. A standardised FACS assay based on native, receptor transfected cells for the clinical diagnosis and monitoring of β1-adrenergic receptor autoantibodies in human heart disease
  30. Infectious Diseases
  31. The diagnostic value of serum fucosylated fetuin A in hepatitis B virus-related liver diseases
  32. Letters to the Editor
  33. Discriminant indices for distinguishing thalassemia and iron deficiency in patients with microcytic anemia
  34. Discriminant indices for distinguishing thalassemia and iron deficiency in patients with microcytic anemia: a reply
  35. STORE (Sample Tracking: Organization, Refrigeration and Expansion): a project in a clinical laboratory perspective
  36. Control materials for immunochromatographic strip used for α-thalassemia screening
  37. Comparison of a 10- vs. 15-min centrifugation time for chemical and immunochemical assays and impact on turnaround time in a hospital laboratory
  38. Pre-analytical phase in cryoglobulin (CRG) detection: an alternative method for sample transport
  39. The efficacy of an internet-based e-learning system using the CellaVision Competency Software for continuing professional development
  40. Clinical examinations after the Fukushima disaster: a case report of Soma General Hospital
  41. Non-fasting plasma glucose concentration in blood donors
  42. Analytical evaluation of a second generation assay for chromogranin A; a dual-site study
  43. UBE3A, c.1347_1348delGA: a mutation in question
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