Prediction of spontaneous autoimmune diabetes in NOD mice by quantification of autoreactive T cells in peripheral blood

Abstract

Type 1 diabetes is an autoimmune disorder characterized by the selective destruction of pancreatic β cells, resulting in insulin deficiency and hyperglycemia. Studies of both humans (1) and the NOD mouse model (2, 3) have shown that β cell destruction is mediated largely by T lymphocytes. Despite the direct β cell cytotoxic role of T cells during diabetes progression, prediction of disease in both humans and the NOD mouse has been based primarily on the presence of circulating autoantibodies to putative T cell antigens (4, 5). Detection and characterization of antigen-specific T cells from peripheral blood during the progression of type 1 diabetes, or other autoimmune diseases, has thus far not been possible without in vitro manipulation. Consequently, little is known regarding the evolution of autoreactive T cell populations during the natural history of autoimmune diseases.Although the detection of antigen-specific T cells using MHC tetramers has permitted careful characterization of T cell responses to many viral and bacterial infections (6, 7), attempts to elucidate the in vivo dynamics of autoreactive T cells during the progression of autoimmune diseases have been less successful. With a single exception (8), attempts to visualize autoreactive T cells in peripheral blood or lymphoid organs ex vivo with MHC tetramers bearing naturally-occurring ligands have failed (9-14), possibly because of the lower avidity that T cells have for self versus foreign peptide/MHC ligands (15, 16). To overcome this limitation, we have engineered high-avidity peptide/MHC tetramers capable of stably interacting with low-avidity autoreactive T cells, thus facilitating their detection. The visualization of lowavidity T cell populations in peripheral blood would provide a simple, minimally invasive method for assessing the presence of autoreactive T cells within infiltrated tissues and might therefore be useful for predicting the development of autoimmune disease.The feasibility of employing a high-avidity MHC tetramer to detect relatively low-avidity autoreactive T cells was investigated using NOD mice, an extensively studied model of human type 1 diabetes (3,17). In these mice, diabetes develops by 16 to 20 weeks of age in approximately 80% of females and is preceded by a prolonged period of mononuclear cell inflammation of the Autoimmune (type 1) diabetes mellitus results from the destruction of insulin-producing pancreatic β cells by T lymphocytes. Prediction of cell-mediated autoimmune diseases by direct detection of autoreactive T cells in peripheral blood has proved elusive, in part because of their low frequency and reduced avidity for peptide MHC ligands. We demonstrate here that MHC class I tetramers complexed to a high-avidity analogue of an immunodominant β cell epitope detect diabetogenic CD8 + T cells in the peripheral blood of NOD mice ex vivo and that the quantification of this autoreactive T cell population in peripheral blood is a powerful predictor of autoimmune diabetes.This article was published online in advan...

Recurrence of autoreactive antigen-specific CD4+ T cells in autoimmune diabetes after pancreas transplantation

“…The variability in detection of peripheral tetramer-binding cells, which was observed over several years in our study, may reflect true changes in cell frequency or, alternatively, changes in the sequestration and circulation of these cells over time. Similar variability has been previously reported in a mouse model of diabetes, using class I tetramer detection strategies [20]. Recently, a study of tetramer-positive cells in celiac disease demonstrated successful detection in peripheral blood samples after antigen challenge in vivo, suggesting the possibility that autoreactive cells may persist in extravascular sites, presumably in the target organ or associated lymphatics, and that they can be mobilized for detection via sampling of the peripheral blood [18].…”

Apportioning Blame: Autoreactive CD4+ and CD8+ T Cells in Type 1 Diabetes

“…For some of these novel autoantigens, there is data suggesting their significant role in the development of T1D, such as the existence of T-cell clones capable of transferring the disease or immunotherapy using antigenic peptides derived from these autoantigens (Table 1). These include islet-specific glusose-6-phosphatase catalytic subunitrelated protein (IGRP), ZnT8 and, interestingly, the neurotropic factor S100beta and the glial fibrillary acidic protein (GFAP) (Gomez-Tourino et al 2010Lieberman et al 2003;Trudeau et al 2003;Winer et al 2003), the latter two being antigens not expressed by the beta cells themselves but by a group of nerve cells surrounding the islets called the peri-islet Schwann cells (pSC) which makes their role in T1D pathogenesis controversial.…”

“…Its relevance to T1D derives from the fact that in the NOD mice the number of IGRP-specific CD8 + T cells correlate with the future development of clinical disease (Trudeau et al 2003).…”

“…Interestingly, these IGRPspecific T cells have a high frequency amongst the isletinfiltrating T lymphocytes (Trudeau et al 2003), where a substantial proportion of CD8 + T cells with a shared T cell receptor (TCR) α chain (Vα17-Jα42) recognize an IGRPderived peptide. The pathogenic role of this prevalent T cell population is well established through the study of the 8.3 T cell clone, representative of this T cell population (Verdaguer et al 1997).…”

Prospective prediction of spontaneous but not recurrent autoimmune diabetes in the non-obese diabetic mouse

“…These CTL recognise peptide sequences from the insulin B chain [15][16][17][18][19][20][21][22][23] (G9C8 clone) [2], islet-specific glucose-6-phosphatase catalytic subunit-related protein (IGRP 206-214 ; 8.3 clone) [3] and dystrophia myotonica kinase (AI4 clone) [4]. Together, these CTL populations constitute a large proportion (up to 60%) of the CD8 + T cells within the pancreatic islets of prediabetic NOD mice [4,5].…”