title: Detection of clonal T-cell-receptor (TCR) Vbeta rearrangements in explanted dilated cardiomyopathy hearts by semi-nested PCR, GeneScan, and direct sequencing.
abstract: BACKGROUND
  Viral infection and anti-cardiac immunity are involved in the pathogenesis of dilated cardiomyopathy (DCM). Immunity targeting particular antigens may evoke expansion of reactive T-cell clones.
  
  
  METHODS
  Myocardial tissues from explanted hearts were investigated for clonal T-cell-receptor- (TCR-) ß rearrangements by an established semi-nested polymerase chain reaction (PCR), followed by high-resolution GeneScan analysis and direct sequencing. From 17 explanted DCM hearts, 3 myocardial samples each were obtained from the right ventricle, the septum, and the left ventricle (total: 9 myocardial samples per case). Six explanted hearts with non-DCM cardiomyopathy entities served as controls.
  
  
  RESULTS
  GeneScan analysis revealed polyclonal TCR-ß rearrangements in all controls. In contrast, at least 1 myocardial sample in 9 out of 17 DCM hearts (total: 20 of the 81 DCM specimens) displayed single dominant TCR-ß PCR products consistent with the presence of clonal T-cell populations. Direct sequencing of the clonal TCR-ß PCR-products disclosed an involvement of Vß 19.01 segments in 14 of the dominant amplificates (70%). Further TCR-Vß segments involved in clonal TCR-ß rearrangements of DCM hearts were Vß 6-1.01 (n=1), Vß 6-3.01 (n=2), Vß 6-5.01 (n=1), Vß 10-3.02 (n=1), and Vß 19.03 (n=1).
  
  
  CONCLUSIONS
  The detectability of clonal TCR-ß rearrangements indicates a pathogenic relevance of this finding in DCM. The predominance of Vß 19.01 segments suggests that the immune response in DCM patients targets particular epitopes. However, the partly heterogenic TCR-ß populations in various myocardial samples from the respective cases support the notion that T-cell immunity may target multiple epitopes in human DCM.
authors.affiliation: Department of Cardiology and Pneumonology, Charité - Universitätsmedizin Berlin, Campus Benjamin Franklin, Berlin, Germany.
authors.forename: Jan-Henrik
authors.initials: JH
authors.lastname: Blohm
authors.forename: Nadine
authors.initials: N
authors.lastname: Blohm
authors.forename: Michael
authors.initials: M
authors.lastname: Hummel
authors.forename: Hans-Henning
authors.initials: HH
authors.lastname: Müller
authors.forename: Maria
authors.initials: M
authors.lastname: Rohde
authors.forename: Roland
authors.initials: R
authors.lastname: Hetzer
authors.forename: Hans
authors.initials: H
authors.lastname: Lehmkuhl
authors.forename: Michel
authors.initials: M
authors.lastname: Noutsias
mesh_terms: D000328:Adult; D000941:Antigens / Q000737:chemistry; D002311:Cardiomyopathy, Dilated / Q000235:genetics / Q000276:immunology* / Q000473:pathology*; D000939:Epitopes / Q000737:chemistry; D005260:Female; D015333:Gene Rearrangement, beta-Chain T-Cell Antigen Receptor / Q000235:genetics*; D006801:Humans; D008297:Male; D008875:Middle Aged; D009205:Myocarditis / Q000235:genetics / Q000276:immunology / Q000473:pathology; D009206:Myocardium / Q000473:pathology; D016133:Polymerase Chain Reaction / Q000379:methods*; D017422:Sequence Analysis, DNA / Q000379:methods*
chemical_list: D000941:Antigens; D000939:Epitopes