Islet-specific CTL cloned from a type 1 diabetes patient cause beta-cell destruction after engraftment into HLA-A2 transgenic NOD/scid/IL2RG null mice

• Published in: PloS one Vol. 7; no. 11; p. e49213
• Main Authors: Unger, Wendy W J, Pearson, Todd, Abreu, Joana R F, Laban, Sandra, van der Slik, Arno R, der Kracht, Sacha Mulder-van, Kester, Michel G D, Serreze, Dave V, Shultz, Leonard D, Griffioen, Marieke, Drijfhout, Jan Wouter, Greiner, Dale L, Roep, Bart O
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 14.11.2012; Public Library of Science (PLoS)
• Subjects: Animals; Antigens; Beta cells; Biology; Blood; Blood transfusions; Catalysis; CD8 antigen; CD8-Positive T-Lymphocytes - immunology; CD8-Positive T-Lymphocytes - metabolism; Cytotoxicity; Destruction; Destructive testing; Diabetes; Diabetes mellitus; Diabetes Mellitus, Type 1 - genetics; Diabetes Mellitus, Type 1 - immunology; Diabetes Mellitus, Type 1 - metabolism; Epitopes; Epitopes - immunology; Epitopes - metabolism; Glucose; Glucose-6-phosphatase; Granzyme B; Hematology; Histocompatibility antigen HLA; HLA-A2 Antigen - genetics; HLA-A2 Antigen - immunology; HLA-A2 Antigen - metabolism; Humans; Insulin-Secreting Cells - immunology; Insulin-Secreting Cells - metabolism; Insulitis; Laboratories; Lymphocytes; Lymphocytes T; Medicine; Mice; Mice, Inbred NOD; Mice, Transgenic; Pancreas; Pathogenesis; Pathogenicity; Pathogens; Peripheral blood; Rodents; Studies; T cell receptors; T-Lymphocytes, Cytotoxic - immunology; T-Lymphocytes, Cytotoxic - metabolism; γ-Interferon; Netherlands; United States > US; Maine; Massachusetts
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0049213

Despite increasing evidence that autoreactive CD8 T-cells are involved in both the initiation of type 1 diabetes (T1D) and the destruction of beta-cells, direct evidence for their destructive role in-vivo is lacking. To address a destructive role for autoreactive CD8 T-cells in human disease, we assessed the pathogenicity of a CD8 T-cell clone derived from a T1D donor and specific for an HLA-A2-restricted epitope of islet-specific glucose-6-phosphatase catalytic-subunit related protein (IGRP). HLA-A2/IGRP tetramer staining revealed a higher frequency of IGRP-specific CD8 T-cells in the peripheral blood of recent onset human individuals than of healthy donors. IGRP(265-273)-specific CD8 T-cells that were cloned from the peripheral blood of a recent onset T1D individual were shown to secrete IFNγ and Granzyme B after antigen-specific activation and lyse HLA-A2-expressing murine islets in-vitro. Lytic capacity was also demonstrated in-vivo by specific killing of peptide-pulsed target cells. Using the HLA-A2 NOD-scid IL2rγ(null) mouse model, HLA-A2-restricted IGRP-specific CD8 T-cells induced a destructive insulitis. Together, this is the first evidence that human HLA-restricted autoreactive CD8 T-cells target HLA-expressing beta-cells in-vivo, demonstrating the translational value of humanized mice to study mechanisms of disease and therapeutic intervention strategies.