title: Detection of GAD65 autoreactive T-cells by HLA class I tetramers in type 1 diabetic patients.
abstract: Type 1 diabetes (T1D) is an autoimmune disease, in which pancreatic beta cells are destroyed in genetically predisposed individuals. While the direct contribution of autoantibodies to the disease pathogenesis is controversial, it is generally recognised that the mechanism of beta cell destruction is mediated by autoreactive T cells that had escaped the thymic selection. We aimed to design a method to detect circulating CD8+ T cells autoreactive against an epitope of the glutamic acid decarboxylase autoantigen, isoform 65 (GAD65) ex vivo in T1D patients by using HLA class I tetramers. Low frequencies of GAD65 peptide-specific CD8+ cytotoxic T lymphocytes were detected in peripheral blood lymphocytes (PBMC) of normal controls after GAD65 peptide-specific stimulation. Conversely, their frequencies were significantly higher than in controls in PBMC of T1D patients after GAD65 peptide stimulation. These preliminary data are encouraging in order to develop a reliable assay to be employed in large-scale screening studies.
authors.affiliation: Autoimmunity and Organ Regeneration Laboratory, Children's Hospital Bambino Gesù, Research Institute (IRCCS), Piazza S. Onofrio 4, 00165 Rome, Italy.
authors.forename: Laura
authors.initials: L
authors.lastname: Giuliani
authors.forename: Raffaella
authors.initials: R
authors.lastname: Mele
authors.forename: Monica
authors.initials: M
authors.lastname: Di Giovine
authors.forename: Laura
authors.initials: L
authors.lastname: Altieri
authors.forename: Antonino
authors.initials: A
authors.lastname: Crinò
authors.forename: Lucilla
authors.initials: L
authors.lastname: Ravà
authors.forename: Alessandra
authors.initials: A
authors.lastname: Fierabracci
mesh_terms: D000293:Adolescent; D001324:Autoantigens / Q000378:metabolism*; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology / Q000378:metabolism*; D002478:Cells, Cultured; D002648:Child; D003922:Diabetes Mellitus, Type 1 / Q000276:immunology / Q000378:metabolism*; D005260:Female; D005434:Flow Cytometry; D005968:Glutamate Decarboxylase / Q000276:immunology / Q000378:metabolism*; D015234:HLA-A Antigens / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D015789:HLA-A2 Antigen; D006801:Humans; D007963:Leukocytes, Mononuclear; D008297:Male; D055503:Protein Multimerization; D018709:Statistics, Nonparametric
chemical_list: D001324:Autoantigens; D015234:HLA-A Antigens; C435939:HLA-A*02:01 antigen; D015789:HLA-A2 Antigen; D005968:Glutamate Decarboxylase