Autoreactive T cell Responses in Insulin-dependent (Type 1) Diabetes Mellitus. Report of the First International Workshop for Standardization of T cell assays

Roep, Bart O.; Atkinson, Mark A.; Endert, Peter Van; Gottlieb, Peter A.; Wilson, S. Brian; Sachs, J. A.

doi:10.1006/jaut.1999.0312

Cited by 120 publications

(82 citation statements)

References 67 publications

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“…Selection of patients based on permanent insulin requirement from the time of diagnosis also limited the potential inclusion of latent autoimmune diabetes in adults (LADA) patients (44). Importantly (35,45,46), detection of ␤-cell-specific CD8 ϩ T-cells was accomplished on both fresh and frozen PBMCs and directly ex vivo. This rules out any bias introduced by in vitro manipulation, while encouraging the transfer to clinical application.…”

Section: Cd8mentioning

confidence: 99%

“…Different assays were compared for their ability to distinguish type 1 diabetes from healthy donors, using 23 frozen samples from new-onset patients. While HLA class II tetramer (36) and IFN-␥ ELISpot assays (49) could not be evaluated because of poor performance on frozen samples (45,46), a cellular immunoblot assay reached 91% sensitivity and 83% specificity (50,51). A T-cell proliferation assay (52) showed a lower sensitivity (58%) but higher specificity (94%).…”

Section: Cd8mentioning

confidence: 99%

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CD8+ T-Cell Responses Identify β-Cell Autoimmunity in Human Type 1 Diabetes

et al. 2007

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Section: Cd8mentioning

confidence: 99%

Section: Cd8mentioning

confidence: 99%

CD8+ T-Cell Responses Identify β-Cell Autoimmunity in Human Type 1 Diabetes

et al. 2007

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“…However, antibodies to these autoantigens do not appear to play a pathogenic role, but T-cells would play crucial roles in destroying the pancreatic ␤-cells in the development of type 1 diabetes (3). Several laboratories have tried to detect cytotoxic T-cells from the peripheral blood of type 1 diabetic patients (6), but conclusive results have yet to be established.…”

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confidence: 99%

Pancreatic Biopsy as a Procedure for Detecting In Situ Autoimmune Phenomena in Type 1 Diabetes

et al. 2001

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To better understand the pathogenesis of type 1 diabetes, we have developed pancreatic biopsy under laparoscope for recent-onset type 1 diabetic patients. The patients included 29 acute-onset type 1 diabetic patients, 5 latent-onset type 1 diabetic patients, and 1 type 2 diabetic patient. Their median age was 28 years, and the duration of diabetes at the time of biopsy was ϳ3 months. In 31 of 35 patients, we could obtain the pancreas tissue by punching. No serious complications, such as heavy bleeding, peritonitis, or pancreatitis, have been experienced. Pneumoderma was observed in two patients, and abdominal dull pain had continued for 2 days in two patients. However, special treatment was not necessary for these complications. T-cell-predominant infiltration to islets (insulitis) and hyperexpression of major histocompatibility complex class I antigens on islet cells were the two major findings and were observed in 17 of 29 recent-onset type 1 diabetic patients. These findings could be regarded as evidence of immune attack against ␤-cells, and their presence was closely correlated with the presence of either anti-GAD or anti-IA-2 antibodies (P ‫؍‬ 0.02). In conclusion, pancreatic biopsy under laparoscope is a safe procedure without serious complications, according to our findings, for detecting in situ autoimmune phenomenon in recent-onset type 1 diabetic patients.

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“…Ultimately, this implies that the use of one specific GAD65 antibody, particularly a polyclonal antibody, may not be sufficient to characterize the binding affinity of a given GAD65 antigen, particularly that of T cell autoreactivity and GAD immunizations [24]. At present, there is no consensus on the quantitative effects of GAD65 on APC in vitro, despite exhaustive attempts by investigators worldwide to standardize GAD65-specific T cell responses [8,9]. Our study suggests that factors other than GAD65 concentrations have to be taken into account when analyzing the ability of GAD65 to stimulate T cells in vitro.…”

Section: Discussionmentioning

confidence: 99%

“…Autoantibodies directed to GAD65 (GAD65Ab) are present in the majority of new onset T1D patients [3,4] and GAD65-specific T cells have been identified in T1D patients and in spontaneously diabetic NOD mice and BB rats (for review see [5,6]). However, while GAD65Ab are readily detectable in radioligand binding assays with good correspondence between different laboratories [7], measurement of T cell responses to GAD65 vary markedly [8] and only some GAD65 preparations have shown reproducible results in international standardization efforts [9]. The establishment of a standard T cell assay remains critical for the understanding of GAD65 antigen uptake, processing and presentation.…”

Section: Introductionmentioning

confidence: 99%

Antigen presentation of detergent-free glutamate decarboxylase (GAD65) is affected by human serum albumin as carrier protein

Steed

Gilliam

Harris³

et al. 2008

Journal of Immunological Methods

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SummaryThe smaller isoform of glutamate decarboxylase (GAD65) is a major autoantigen in type 1 diabetes (TID). Its hydrophobic character requires detergent to keep the protein in solution, which complicates studies of antigen processing and presentation. In this study an attempt was made to replace detergent with human serum albumin (HSA) for in vitro antigen presentation. Different preparations of recombinant human GAD65 complexed with HSA were incubated with Priess B cells (HLA DRB1*0401) and antigen presentation was tested with HLA DRB1*0401-restricted and epitopespecific T33.1 (GAD65 epitope 274-286) and T35 (GAD65 epitope 115-127) T cell hybridomas. Specific epitope recognition by T33.1 (274-286) and T35 (115-127) cells varied between the different GAD65/HSA preparations, and a reverse pattern of antigen presentation were detected by the two hybridoma. The HSA-specific T-cell hybridoma 17.9 response to the different GAD65/HSA preparations followed the same pattern as that observed for the T33.1 cells. The content of immunoreactive GAD65 measured with four GAD65 antibodies indicated that the lowest GAD65 concentration resulted in the highest 274-286, but the lowest 115-127 presentation. This suggests that HSA-GAD65 complexes qualitatively affect the epitope specificity of GAD65 presentation. HSA may enhance the 274-286 epitope presentation, while suppressing the 115-127 epitope.Keywords antigen presentation; GAD65; Human serum albumin; T cell assay; Type 1 diabetes; T cells; Diabetes; Autoimmunity; Antigen Presentation/Processing IntroductionType 1 diabetes mellitus (TID) is an autoimmune disease that results in the specific destruction of the pancreatic islet beta cells. The smaller isoform of glutamate decarboxylase (GAD65) is implicated as a major contributing autoantigen in the pathogenesis of beta cell destruction [1,2]. Autoantibodies directed to GAD65 (GAD65Ab) are present in the majority of new onset T1D patients [3,4] and GAD65-specific T cells have been identified in T1D patients and in spontaneously diabetic NOD mice and BB rats (for review see [5,6] [9]. The establishment of a standard T cell assay remains critical for the understanding of GAD65 antigen uptake, processing and presentation.GAD65, which catalyzes the generation of the neurotransmitter GABA, is associated with intracellular membranes [10,11]. In vitro, the highly hydrophobic GAD65 requires detergents to remain in solution [12]. However, because detergents are extremely cytotoxic, it is critical that the detergent is removed prior to incubation with antigen-presenting cells (APC) and responder T cells.In the present study, we tested the well known property of human serum albumin (HSA) to maintain proteins of hydrophobic character in solution (for review see [13,14]). The aim of our study was to test whether GAD65 complexed with HSA affected antigen presentation by human Priess B cells to two GAD65-and one HSA-specific specific HLA-matched T cell hybridoma as readouts of antigen presentation. The specific epitope response of the two ...

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Autoreactive T cell Responses in Insulin-dependent (Type 1) Diabetes Mellitus. Report of the First International Workshop for Standardization of T cell assays

Cited by 120 publications

References 67 publications

CD8+ T-Cell Responses Identify β-Cell Autoimmunity in Human Type 1 Diabetes

CD8+ T-Cell Responses Identify β-Cell Autoimmunity in Human Type 1 Diabetes

Pancreatic Biopsy as a Procedure for Detecting In Situ Autoimmune Phenomena in Type 1 Diabetes

Antigen presentation of detergent-free glutamate decarboxylase (GAD65) is affected by human serum albumin as carrier protein

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