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Journal of Biomedicine and Biotechnology
Volume 2009, Article ID 576219, 7 pages
http://dx.doi.org/10.1155/2009/576219
Methodology Report

Detection of GAD65 Autoreactive T-Cells by HLA Class I Tetramers in Type 1 Diabetic Patients

1Autoimmunity and Organ Regeneration Laboratory, Children's Hospital Bambino Gesù, Research Institute (IRCCS), Piazza S. Onofrio 4, 00165 Rome, Italy
2Unit of Pediatric Autoimmune Endocrine Diseases, Children's Hospital Bambino Gesù, Research Institute (IRCCS), Piazza S. Onofrio 4, 00165 Rome, Italy
3Department of Epidemiology, Children's Hospital Bambino Gesù, Research Institute (IRCCS), Piazza S. Onofrio 4, 00165 Rome, Italy

Received 23 April 2009; Revised 30 July 2009; Accepted 30 August 2009

Academic Editor: Marija Mostarica-Stojković

Copyright © 2009 Laura Giuliani et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Type 1 diabetes (T1D) is an autoimmune disease, in which pancreatic 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 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.