The Experts below are selected from a list of 34956 Experts worldwide ranked by ideXlab platform
Vijay K Kuchroo - One of the best experts on this subject based on the ideXlab platform.
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detection of autoreactive myelin proteolipid protein 139 151 specific t cells by using mhc ii ias Tetramers
Journal of Immunology, 2003Co-Authors: N Jayagopala R Reddy, Lindsay B Nicholson, Estelle Bettelli, Kai W Wucherpfennig, Hanspeter Waldner, Mei Huei Jang, Vijay K KuchrooAbstract:Detection of autoreactive T cells using MHC II Tetramers is difficult because of the low affinity of their TCR. We have generated a class II Tetramer using the IA s class II molecule combined with an autoantigenic peptide from myelin proteolipid protein (PLP; PLP 139–151 ) and used it to analyze myelin PLP 139–151 -reactive T cells. Using monomers and multimerized complexes labeled with PE, we confirmed the specificity of the reagent by bioassay and flow cytometry. The IA s Tetramers stimulated and stained the PLP 139–151 -specific 5B6 TCR transgenic T cells and a polyclonal cell line specific for PLP 139–151 , but not a control T cell line specific for PLP 178–191 . We used this reagent to optimize conditions to detect low affinity autoreactive T cells. We found that high pH (∼8.0) and neuraminidase treatment enhances the staining capacity of PLP 139–151 Tetramer without compromising specificity. Furthermore, we found that induction of calcium fluxing by Tetramers in T cells may be used as a sensitive measure to detect autoreactive T cells with a low affinity. Taken together, the data show that the Tetrameric reagent binds and stimulates PLP 139–151 -reactive T cells with specificity. This Tetrameric reagent will be useful in studying the evolution of PLP 139–151 -specific repertoire in naive mice and its expansion during the autoimmune disease experimental autoimmune encephalomyelitis.
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detection of autoreactive myelin proteolipid protein 139 151 specific t cells by using mhc ii ias Tetramers
Journal of Immunology, 2003Co-Authors: N Jayagopala R Reddy, Lindsay B Nicholson, Estelle Bettelli, Kai W Wucherpfennig, Hanspeter Waldner, Mei Huei Jang, Vijay K KuchrooAbstract:Detection of autoreactive T cells using MHC II Tetramers is difficult because of the low affinity of their TCR. We have generated a class II Tetramer using the IA(s) class II molecule combined with an autoantigenic peptide from myelin proteolipid protein (PLP; PLP(139-151)) and used it to analyze myelin PLP(139-151)-reactive T cells. Using monomers and multimerized complexes labeled with PE, we confirmed the specificity of the reagent by bioassay and flow cytometry. The IA(s) Tetramers stimulated and stained the PLP(139-151)-specific 5B6 TCR transgenic T cells and a polyclonal cell line specific for PLP(139-151), but not a control T cell line specific for PLP(178-191). We used this reagent to optimize conditions to detect low affinity autoreactive T cells. We found that high pH ( approximately 8.0) and neuraminidase treatment enhances the staining capacity of PLP(139-151) Tetramer without compromising specificity. Furthermore, we found that induction of calcium fluxing by Tetramers in T cells may be used as a sensitive measure to detect autoreactive T cells with a low affinity. Taken together, the data show that the Tetrameric reagent binds and stimulates PLP(139-151)-reactive T cells with specificity. This Tetrameric reagent will be useful in studying the evolution of PLP(139-151)-specific repertoire in naive mice and its expansion during the autoimmune disease experimental autoimmune encephalomyelitis.
Kai W Wucherpfennig - One of the best experts on this subject based on the ideXlab platform.
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ex vivo analysis of thymic cd4 t cells in nonobese diabetic mice with Tetramers generated from i ag7 class ii associated invariant chain peptide precursors
Journal of Immunology, 2003Co-Authors: Mei Huei Jang, Nilufer P Seth, Kai W WucherpfennigAbstract:The MHC determines susceptibility and resistance to type 1 diabetes in humans and nonobese diabetic (NOD) mice. To investigate how a disease-associated MHC molecule shapes the T cell repertoire in NOD mice, we generated a series of Tetramers from I-Ag7/class II-associated invariant chain peptide precursors by peptide exchange. No CD4 T cell populations could be identified for two glutamic acid decarboxylase 65 peptides, but Tetramers with a peptide mimetic recognized by the BDC-2.5 and other islet-specific T cell clones labeled a distinct population in the thymus of young NOD mice. Tetramer-positive cells were identified in the immature CD4+CD8low population that arises during positive selection, and in larger numbers in the more mature CD4+CD8− population. Tetramer labeling was specific based on the use of multiple control Tetramers, including one with a single amino acid analog peptide in which a critical TCR contact residue was substituted. The T cell population was already present in the thymus of 2-wk-old NOD mice before the typical onset of insulitis and was detected in B10 mice congenic for the NOD MHC locus, but not B10 control mice. These results demonstrate that a T cell population can expand in the thymus of NOD mice to levels that are at least two to three orders of magnitude higher than estimated for a given specificity in the naive T cell pool. Based on these data, we propose a model in which I-Ag7 confers susceptibility to type 1 diabetes by biasing positive selection in the thymus and later presenting peptides from islet autoantigens to such T cells in the periphery.
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detection of autoreactive myelin proteolipid protein 139 151 specific t cells by using mhc ii ias Tetramers
Journal of Immunology, 2003Co-Authors: N Jayagopala R Reddy, Lindsay B Nicholson, Estelle Bettelli, Kai W Wucherpfennig, Hanspeter Waldner, Mei Huei Jang, Vijay K KuchrooAbstract:Detection of autoreactive T cells using MHC II Tetramers is difficult because of the low affinity of their TCR. We have generated a class II Tetramer using the IA s class II molecule combined with an autoantigenic peptide from myelin proteolipid protein (PLP; PLP 139–151 ) and used it to analyze myelin PLP 139–151 -reactive T cells. Using monomers and multimerized complexes labeled with PE, we confirmed the specificity of the reagent by bioassay and flow cytometry. The IA s Tetramers stimulated and stained the PLP 139–151 -specific 5B6 TCR transgenic T cells and a polyclonal cell line specific for PLP 139–151 , but not a control T cell line specific for PLP 178–191 . We used this reagent to optimize conditions to detect low affinity autoreactive T cells. We found that high pH (∼8.0) and neuraminidase treatment enhances the staining capacity of PLP 139–151 Tetramer without compromising specificity. Furthermore, we found that induction of calcium fluxing by Tetramers in T cells may be used as a sensitive measure to detect autoreactive T cells with a low affinity. Taken together, the data show that the Tetrameric reagent binds and stimulates PLP 139–151 -reactive T cells with specificity. This Tetrameric reagent will be useful in studying the evolution of PLP 139–151 -specific repertoire in naive mice and its expansion during the autoimmune disease experimental autoimmune encephalomyelitis.
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detection of autoreactive myelin proteolipid protein 139 151 specific t cells by using mhc ii ias Tetramers
Journal of Immunology, 2003Co-Authors: N Jayagopala R Reddy, Lindsay B Nicholson, Estelle Bettelli, Kai W Wucherpfennig, Hanspeter Waldner, Mei Huei Jang, Vijay K KuchrooAbstract:Detection of autoreactive T cells using MHC II Tetramers is difficult because of the low affinity of their TCR. We have generated a class II Tetramer using the IA(s) class II molecule combined with an autoantigenic peptide from myelin proteolipid protein (PLP; PLP(139-151)) and used it to analyze myelin PLP(139-151)-reactive T cells. Using monomers and multimerized complexes labeled with PE, we confirmed the specificity of the reagent by bioassay and flow cytometry. The IA(s) Tetramers stimulated and stained the PLP(139-151)-specific 5B6 TCR transgenic T cells and a polyclonal cell line specific for PLP(139-151), but not a control T cell line specific for PLP(178-191). We used this reagent to optimize conditions to detect low affinity autoreactive T cells. We found that high pH ( approximately 8.0) and neuraminidase treatment enhances the staining capacity of PLP(139-151) Tetramer without compromising specificity. Furthermore, we found that induction of calcium fluxing by Tetramers in T cells may be used as a sensitive measure to detect autoreactive T cells with a low affinity. Taken together, the data show that the Tetrameric reagent binds and stimulates PLP(139-151)-reactive T cells with specificity. This Tetrameric reagent will be useful in studying the evolution of PLP(139-151)-specific repertoire in naive mice and its expansion during the autoimmune disease experimental autoimmune encephalomyelitis.
Mei Huei Jang - One of the best experts on this subject based on the ideXlab platform.
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ex vivo analysis of thymic cd4 t cells in nonobese diabetic mice with Tetramers generated from i ag7 class ii associated invariant chain peptide precursors
Journal of Immunology, 2003Co-Authors: Mei Huei Jang, Nilufer P Seth, Kai W WucherpfennigAbstract:The MHC determines susceptibility and resistance to type 1 diabetes in humans and nonobese diabetic (NOD) mice. To investigate how a disease-associated MHC molecule shapes the T cell repertoire in NOD mice, we generated a series of Tetramers from I-Ag7/class II-associated invariant chain peptide precursors by peptide exchange. No CD4 T cell populations could be identified for two glutamic acid decarboxylase 65 peptides, but Tetramers with a peptide mimetic recognized by the BDC-2.5 and other islet-specific T cell clones labeled a distinct population in the thymus of young NOD mice. Tetramer-positive cells were identified in the immature CD4+CD8low population that arises during positive selection, and in larger numbers in the more mature CD4+CD8− population. Tetramer labeling was specific based on the use of multiple control Tetramers, including one with a single amino acid analog peptide in which a critical TCR contact residue was substituted. The T cell population was already present in the thymus of 2-wk-old NOD mice before the typical onset of insulitis and was detected in B10 mice congenic for the NOD MHC locus, but not B10 control mice. These results demonstrate that a T cell population can expand in the thymus of NOD mice to levels that are at least two to three orders of magnitude higher than estimated for a given specificity in the naive T cell pool. Based on these data, we propose a model in which I-Ag7 confers susceptibility to type 1 diabetes by biasing positive selection in the thymus and later presenting peptides from islet autoantigens to such T cells in the periphery.
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detection of autoreactive myelin proteolipid protein 139 151 specific t cells by using mhc ii ias Tetramers
Journal of Immunology, 2003Co-Authors: N Jayagopala R Reddy, Lindsay B Nicholson, Estelle Bettelli, Kai W Wucherpfennig, Hanspeter Waldner, Mei Huei Jang, Vijay K KuchrooAbstract:Detection of autoreactive T cells using MHC II Tetramers is difficult because of the low affinity of their TCR. We have generated a class II Tetramer using the IA s class II molecule combined with an autoantigenic peptide from myelin proteolipid protein (PLP; PLP 139–151 ) and used it to analyze myelin PLP 139–151 -reactive T cells. Using monomers and multimerized complexes labeled with PE, we confirmed the specificity of the reagent by bioassay and flow cytometry. The IA s Tetramers stimulated and stained the PLP 139–151 -specific 5B6 TCR transgenic T cells and a polyclonal cell line specific for PLP 139–151 , but not a control T cell line specific for PLP 178–191 . We used this reagent to optimize conditions to detect low affinity autoreactive T cells. We found that high pH (∼8.0) and neuraminidase treatment enhances the staining capacity of PLP 139–151 Tetramer without compromising specificity. Furthermore, we found that induction of calcium fluxing by Tetramers in T cells may be used as a sensitive measure to detect autoreactive T cells with a low affinity. Taken together, the data show that the Tetrameric reagent binds and stimulates PLP 139–151 -reactive T cells with specificity. This Tetrameric reagent will be useful in studying the evolution of PLP 139–151 -specific repertoire in naive mice and its expansion during the autoimmune disease experimental autoimmune encephalomyelitis.
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detection of autoreactive myelin proteolipid protein 139 151 specific t cells by using mhc ii ias Tetramers
Journal of Immunology, 2003Co-Authors: N Jayagopala R Reddy, Lindsay B Nicholson, Estelle Bettelli, Kai W Wucherpfennig, Hanspeter Waldner, Mei Huei Jang, Vijay K KuchrooAbstract:Detection of autoreactive T cells using MHC II Tetramers is difficult because of the low affinity of their TCR. We have generated a class II Tetramer using the IA(s) class II molecule combined with an autoantigenic peptide from myelin proteolipid protein (PLP; PLP(139-151)) and used it to analyze myelin PLP(139-151)-reactive T cells. Using monomers and multimerized complexes labeled with PE, we confirmed the specificity of the reagent by bioassay and flow cytometry. The IA(s) Tetramers stimulated and stained the PLP(139-151)-specific 5B6 TCR transgenic T cells and a polyclonal cell line specific for PLP(139-151), but not a control T cell line specific for PLP(178-191). We used this reagent to optimize conditions to detect low affinity autoreactive T cells. We found that high pH ( approximately 8.0) and neuraminidase treatment enhances the staining capacity of PLP(139-151) Tetramer without compromising specificity. Furthermore, we found that induction of calcium fluxing by Tetramers in T cells may be used as a sensitive measure to detect autoreactive T cells with a low affinity. Taken together, the data show that the Tetrameric reagent binds and stimulates PLP(139-151)-reactive T cells with specificity. This Tetrameric reagent will be useful in studying the evolution of PLP(139-151)-specific repertoire in naive mice and its expansion during the autoimmune disease experimental autoimmune encephalomyelitis.
N Jayagopala R Reddy - One of the best experts on this subject based on the ideXlab platform.
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detection of autoreactive myelin proteolipid protein 139 151 specific t cells by using mhc ii ias Tetramers
Journal of Immunology, 2003Co-Authors: N Jayagopala R Reddy, Lindsay B Nicholson, Estelle Bettelli, Kai W Wucherpfennig, Hanspeter Waldner, Mei Huei Jang, Vijay K KuchrooAbstract:Detection of autoreactive T cells using MHC II Tetramers is difficult because of the low affinity of their TCR. We have generated a class II Tetramer using the IA s class II molecule combined with an autoantigenic peptide from myelin proteolipid protein (PLP; PLP 139–151 ) and used it to analyze myelin PLP 139–151 -reactive T cells. Using monomers and multimerized complexes labeled with PE, we confirmed the specificity of the reagent by bioassay and flow cytometry. The IA s Tetramers stimulated and stained the PLP 139–151 -specific 5B6 TCR transgenic T cells and a polyclonal cell line specific for PLP 139–151 , but not a control T cell line specific for PLP 178–191 . We used this reagent to optimize conditions to detect low affinity autoreactive T cells. We found that high pH (∼8.0) and neuraminidase treatment enhances the staining capacity of PLP 139–151 Tetramer without compromising specificity. Furthermore, we found that induction of calcium fluxing by Tetramers in T cells may be used as a sensitive measure to detect autoreactive T cells with a low affinity. Taken together, the data show that the Tetrameric reagent binds and stimulates PLP 139–151 -reactive T cells with specificity. This Tetrameric reagent will be useful in studying the evolution of PLP 139–151 -specific repertoire in naive mice and its expansion during the autoimmune disease experimental autoimmune encephalomyelitis.
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detection of autoreactive myelin proteolipid protein 139 151 specific t cells by using mhc ii ias Tetramers
Journal of Immunology, 2003Co-Authors: N Jayagopala R Reddy, Lindsay B Nicholson, Estelle Bettelli, Kai W Wucherpfennig, Hanspeter Waldner, Mei Huei Jang, Vijay K KuchrooAbstract:Detection of autoreactive T cells using MHC II Tetramers is difficult because of the low affinity of their TCR. We have generated a class II Tetramer using the IA(s) class II molecule combined with an autoantigenic peptide from myelin proteolipid protein (PLP; PLP(139-151)) and used it to analyze myelin PLP(139-151)-reactive T cells. Using monomers and multimerized complexes labeled with PE, we confirmed the specificity of the reagent by bioassay and flow cytometry. The IA(s) Tetramers stimulated and stained the PLP(139-151)-specific 5B6 TCR transgenic T cells and a polyclonal cell line specific for PLP(139-151), but not a control T cell line specific for PLP(178-191). We used this reagent to optimize conditions to detect low affinity autoreactive T cells. We found that high pH ( approximately 8.0) and neuraminidase treatment enhances the staining capacity of PLP(139-151) Tetramer without compromising specificity. Furthermore, we found that induction of calcium fluxing by Tetramers in T cells may be used as a sensitive measure to detect autoreactive T cells with a low affinity. Taken together, the data show that the Tetrameric reagent binds and stimulates PLP(139-151)-reactive T cells with specificity. This Tetrameric reagent will be useful in studying the evolution of PLP(139-151)-specific repertoire in naive mice and its expansion during the autoimmune disease experimental autoimmune encephalomyelitis.
Hanspeter Waldner - One of the best experts on this subject based on the ideXlab platform.
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detection of autoreactive myelin proteolipid protein 139 151 specific t cells by using mhc ii ias Tetramers
Journal of Immunology, 2003Co-Authors: N Jayagopala R Reddy, Lindsay B Nicholson, Estelle Bettelli, Kai W Wucherpfennig, Hanspeter Waldner, Mei Huei Jang, Vijay K KuchrooAbstract:Detection of autoreactive T cells using MHC II Tetramers is difficult because of the low affinity of their TCR. We have generated a class II Tetramer using the IA s class II molecule combined with an autoantigenic peptide from myelin proteolipid protein (PLP; PLP 139–151 ) and used it to analyze myelin PLP 139–151 -reactive T cells. Using monomers and multimerized complexes labeled with PE, we confirmed the specificity of the reagent by bioassay and flow cytometry. The IA s Tetramers stimulated and stained the PLP 139–151 -specific 5B6 TCR transgenic T cells and a polyclonal cell line specific for PLP 139–151 , but not a control T cell line specific for PLP 178–191 . We used this reagent to optimize conditions to detect low affinity autoreactive T cells. We found that high pH (∼8.0) and neuraminidase treatment enhances the staining capacity of PLP 139–151 Tetramer without compromising specificity. Furthermore, we found that induction of calcium fluxing by Tetramers in T cells may be used as a sensitive measure to detect autoreactive T cells with a low affinity. Taken together, the data show that the Tetrameric reagent binds and stimulates PLP 139–151 -reactive T cells with specificity. This Tetrameric reagent will be useful in studying the evolution of PLP 139–151 -specific repertoire in naive mice and its expansion during the autoimmune disease experimental autoimmune encephalomyelitis.
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detection of autoreactive myelin proteolipid protein 139 151 specific t cells by using mhc ii ias Tetramers
Journal of Immunology, 2003Co-Authors: N Jayagopala R Reddy, Lindsay B Nicholson, Estelle Bettelli, Kai W Wucherpfennig, Hanspeter Waldner, Mei Huei Jang, Vijay K KuchrooAbstract:Detection of autoreactive T cells using MHC II Tetramers is difficult because of the low affinity of their TCR. We have generated a class II Tetramer using the IA(s) class II molecule combined with an autoantigenic peptide from myelin proteolipid protein (PLP; PLP(139-151)) and used it to analyze myelin PLP(139-151)-reactive T cells. Using monomers and multimerized complexes labeled with PE, we confirmed the specificity of the reagent by bioassay and flow cytometry. The IA(s) Tetramers stimulated and stained the PLP(139-151)-specific 5B6 TCR transgenic T cells and a polyclonal cell line specific for PLP(139-151), but not a control T cell line specific for PLP(178-191). We used this reagent to optimize conditions to detect low affinity autoreactive T cells. We found that high pH ( approximately 8.0) and neuraminidase treatment enhances the staining capacity of PLP(139-151) Tetramer without compromising specificity. Furthermore, we found that induction of calcium fluxing by Tetramers in T cells may be used as a sensitive measure to detect autoreactive T cells with a low affinity. Taken together, the data show that the Tetrameric reagent binds and stimulates PLP(139-151)-reactive T cells with specificity. This Tetrameric reagent will be useful in studying the evolution of PLP(139-151)-specific repertoire in naive mice and its expansion during the autoimmune disease experimental autoimmune encephalomyelitis.
