Naturally processed T cell epitopes from human glutamic acid decarboxylase identified using mice transgenic for the type 1 diabetes-associated human MHC class II allele, DRB1*0401.

Wicker, Linda S.; Chen, Shiow Ling; Nepom, Gerald T.; Elliott, John F.; Freed, Daniel C.; Bansal, Alka; Zheng, Song Guo; Herman, Andrew; Lernmark, Åke; Zaller, Dennis M.; Peterson, Laurence B.; Rothbard, Jonathan B.; Cummings, Richard; Whiteley, Phyllis Jonas

doi:10.1172/jci119079

Cited by 133 publications

(116 citation statements)

References 24 publications

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“…The choice of these immunogenic epitopes from among the often hundreds or thousands of amino acids comprising an antigenic protein depends significantly on the binding properties of a given MHC type and the interactions of specific amino acids with the TCR. Understanding which peptide epitopes participate in T cell-mediated immunity provides a basis for directed modulation of the immune response, including development of peptide vaccines and therapies against allergens, autoimmune diseases and tumors (1)(2)(3)(4)(5). However, elucidation of specific epitopes from complex Ags can be a cumbersome and difficult process, as it generally involves extensive phenotype screening of T cell clones isolated from whole Ag-stimulated cells.…”

Section: Tetramer-guided Epitopementioning

confidence: 99%

Tetramer-Guided Epitope Mapping: Rapid Identification and Characterization of Immunodominant CD4+ T Cell Epitopes from Complex Antigens

Novak

Liu²,

Gebe³

et al. 2001

The Journal of Immunology

129

107

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T cell responses to Ags involve recognition of selected peptide epitopes contained within the antigenic protein. In this report, we describe a new approach for direct identification of CD4+ T cell epitopes of complex Ags that uses human class II tetramers to identify reactive cells. With a panel of 60 overlapping peptides covering the entire sequence of the VP16 protein, a major Ag for HSV-2, we generated a panel of class II MHC tetramers loaded with peptide pools that were used to stain peripheral lymphocytes of an HSV-2 infected individual. With this approach, we identified four new DRA1*0101/DRB1*0401- and two DRA1*0101/DRB1*0404-restricted, VP16-specific epitopes. By using tetramers to sort individual cells, we easily obtained a large number of clones specific to these epitopes. Although DRA1*0101/DRB1*0401 and DRA1*0101/DRB1*0404 are structurally very similar, nonoverlapping VP16 epitopes were identified, illustrating high selectivity of individual allele polymorphisms within common MHC variants. This rapid approach to detecting CD4+ T cell epitopes from complex Ags can be applied to any known Ag that gives a T cell response.

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Section: Tetramer-guided Epitopementioning

confidence: 99%

Tetramer-Guided Epitope Mapping: Rapid Identification and Characterization of Immunodominant CD4+ T Cell Epitopes from Complex Antigens

Novak

Liu²,

Gebe³

et al. 2001

The Journal of Immunology

129

107

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“…Although CD4+ T cells expressing TCR that recognize autoantigenpeptides presented on HLA-DR or -DQ heterodimers remain in undisputable target for prevention, it seems equally important that B cells expressing BCR reactive an islet autoantigen are targeted ( Figure 6). As shown in ex vivo experiments, B cells are effective antigen presenting cells 58,59 . Recent studies in the mouse provide evidence that there is a so-called cross-talk between B lymphocytes and CD8+effector T cells that may lead to diabetes (ref).…”

Section: E Future Directionsmentioning

confidence: 92%

Immune therapy in type 1 diabetes mellitus

Lernmark

Larsson

2013

Nat Rev Endocrinol

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(200 words) Type 1 diabetes mellitus (T1D) is an autoimmune disorder directed against the pancreatic islet β cells. The genetic risk for the disease is linked to HLA-DQ genotypes and unknown environmental triggers. In most countries, only 10-15% of newly diagnosed T1D children or young adults have a first degree relative with the disease. Autoantibodies against insulin, GAD65, IA-2 or ZnT8 transporter mark islet autoimmunity. These islet autoantibodies may develop already at 1-2 years of age. Immune therapy in T1D is approached at three stages. Primary prevention is treatment of subjects at increased genetic risk. The TRIGR trial is testing if hydrolyzed casein milk formula may reduce T1D in genetically predisposed infants. Secondary prevention is in subjects with persistent islet autoantibodies. On-going trials are either non-autoantigen (BCG, CD3 monoclonal antibodies) or autoantigen (oral and nasal insulin or Alum-formulated GAD65) specific. Intervention at diabetes onset include non-autoantigen (CD3 monoclonal antibodies, IL-2 receptor antibodies, Il-1b receptor inhibitor, Il-1b antibodies, BCG, ATG, DiaPep277) and autoantigen (proinsulin peptides) specific therapy. Although preserved beta-cell function long term has been difficult to achieve in many prior studies, considerable progress is being made through controlled clinical trials and animal investigations to uncover mechanisms of beta-cell destruction. Novel therapies that would prevent islet autoimmunity or halt progressive beta-cell destruction need to be designed.

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“…GAD65-specific murine T-cell hybridomas T33.1 (274-286) and T35 (115-127) were kindly donated by Dr. Linda Wicker (University of Cambridge, Cambridge, U.K.). The T33.1 (274-286) and T35 (115-127) cells, generated in an HLA-DR4 (DRB1*0401) transgenic mouse, recognized the GAD65 274-286 and 115-127 epitopes, respectively [1]. The T cell hybridomas were maintained in RPMI 1640 and 10% FCS, 1.0 μM ß-mercaptoethanol (BioRad, Hercules, CA), 1 mmol/L sodium pyruvate, 100 U/ml penicillin, and 100 μg/ml streptomycin.…”

Section: Methodsmentioning

confidence: 99%

“…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].…”

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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Naturally processed T cell epitopes from human glutamic acid decarboxylase identified using mice transgenic for the type 1 diabetes-associated human MHC class II allele, DRB1*0401.

Cited by 133 publications

References 24 publications

Tetramer-Guided Epitope Mapping: Rapid Identification and Characterization of Immunodominant CD4+ T Cell Epitopes from Complex Antigens

Tetramer-Guided Epitope Mapping: Rapid Identification and Characterization of Immunodominant CD4+ T Cell Epitopes from Complex Antigens

Immune therapy in type 1 diabetes mellitus

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

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