title: Quantification and characterization of myosin peptide-specific CD4+ T cells in autoimmune myocarditis.
abstract: Characterization of autoantigen-specific CD4+ T cells at the single cell level is crucial for understanding the immunopathological mechanisms underlying autoimmune diseases. Cardiac myosin heavy chain (myhca) is the major autoantigen associated with autoimmune myocarditis both in humans and in experimental autoimmune myocarditis (EAM) in mice. In the current study, we evaluated two methods for the enumeration and phenotypic characterization of myhca-specific CD4+ T cells during the course of EAM. Both enzyme-linked immunospot (ELISPOT) and cytokine flow cytometry (CFC) assays were suitable for the detection and characterization of myhca-specific Th cells during acute myocardial inflammation and the late healing phase of the disease. Cytokine production of myhca-specific Th cells was restricted to interferon-gamma (IFNgamma). Only trace amounts of the Th2 cytokines IL-4 and IL-5 could be detected. Concomitant surface marker analysis in the CFC assay revealed the prototypical effector phenotype of myhca-specific Th1 cells during the acute phase of the disease. Taken together, the combination of both methods appears to be most appropriate for a comprehensive ex vivo single cell analysis of Th cells in heart-specific autoimmune disorders.
authors.affiliation: Research Department, Kantonal Hospital St. Gallen, 9007 St. Gallen, Switzerland.
authors.forename: Reinhard
authors.initials: R
authors.lastname: Maier
authors.forename: Simone
authors.initials: S
authors.lastname: Miller
authors.forename: Michael
authors.initials: M
authors.lastname: Kurrer
authors.forename: Philippe
authors.initials: P
authors.lastname: Krebs
authors.forename: Rita
authors.initials: R
authors.lastname: de Giuli
authors.forename: Marcel
authors.initials: M
authors.lastname: Kremer
authors.forename: Elke
authors.initials: E
authors.lastname: Scandella
authors.forename: Burkhard
authors.initials: B
authors.lastname: Ludewig
mesh_terms: D000818:Animals; D001324:Autoantigens / Q000737:chemistry / Q000276:immunology; D001327:Autoimmune Diseases / Q000276:immunology* / Q000378:metabolism; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology* / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D018984:Epitopes, T-Lymphocyte / Q000737:chemistry / Q000276:immunology*; D005260:Female; D005434:Flow Cytometry / Q000379:methods; D016130:Immunophenotyping / Q000379:methods; D007700:Kinetics; D051379:Mice; D008807:Mice, Inbred BALB C; D009205:Myocarditis / Q000276:immunology*; D018995:Myosin Heavy Chains / Q000737:chemistry / Q000276:immunology*; D010446:Peptide Fragments / Q000737:chemistry / Q000276:immunology*; D018417:Th1 Cells / Q000276:immunology
chemical_list: D001324:Autoantigens; D018984:Epitopes, T-Lymphocyte; D010446:Peptide Fragments; D018995:Myosin Heavy Chains