Antigen processing and presentation, as a field, explores a broad range of protein interactions and functions, both intracellular (in the cytoplasm and in the endoplasmic reticulum) and at the cell surface (between T cells and MHC molecules). To investigate such a diverse array, it is necessary that biochemical, cell biology, and immunological techniques all be employed. The purpose of Antigen Processing and Presentation Protocols is therefore to detail the most up-to-date techniques being used in this burgeoning field. Such techniques include those used to question how MHC-binding peptides…mehr
Antigen processing and presentation, as a field, explores a broad range of protein interactions and functions, both intracellular (in the cytoplasm and in the endoplasmic reticulum) and at the cell surface (between T cells and MHC molecules). To investigate such a diverse array, it is necessary that biochemical, cell biology, and immunological techniques all be employed. The purpose of Antigen Processing and Presentation Protocols is therefore to detail the most up-to-date techniques being used in this burgeoning field. Such techniques include those used to question how MHC-binding peptides are generated, to test how peptides are delivered to MHC molecules, to analyze MHC peptide-binding patterns, and to assay the T-cell response to MHC/peptide. Antigen Processing and Presentation Protocols should aid both those new and those experienced in this area of research in extending the questions that can be asked and answered by the application of these current methods. For editorial assistance, I would like to thank Angela Beninga and Rachael Turnquist.
Purification of 20S Proteasomes.- Use of Proteasome Inhibitors to Examine Processing of Antigens for Major Histocompatibility Complex Class I Presentation.- Tracing the Route Taken by Peptides and Major Histocompatibility Complex Class I Molecules in Presentation of Exogenous Antigens.- Functional Analysis of Antigen Processing and Major Histocompatibility Complex Class II-Restricted Presentation.- Quantitation of Major Histocompatibility Complex Antigen Endocytosis Using Capture ELISA.- In Vitro Generation of Functional Human and Murine Dendritic Cells.- Genetic Engineering of Dendritic Cells Using Retrovirus-Based Gene Transfer Techniques.- Use of Vaccinia Virus Expression Vectors to Investigate Antigen Processing and Presentation.- Sindbis Virus-Based Vectors for the Study of Class I Antigen Presentation In Vitro and In Vivo.- Precision Genotyping of Human Leukocyte Antigen-A, -B and -C Loci Via Direct DNA Sequencing.- Photolabeling the Transporter Associated with Antigen Processing.- Application of One-Dimensional Isoelectric Focusing to Separate Different Major Histocompatibility Complex Class I Alleles and Determine Their Allelic Interactions with Transporter Associated with Antigen Processing (TAP).- Analysis of Major Histocompatibility Complex Class I Interactions with Endoplasmic Reticulum Proteins.- Methods of Purification of Heat Shock Protein-Peptide Complexes for Use as Vaccines Against Cancers and Infectious Diseases.- Random Peptide Libraries.- Neural Network Method for Predicting Peptides that Bind Major Histocompatibility Complex Molecules.- Analysis of T-Cell Response Using Altered Peptide Ligands.- Assays of Thymic Selection Fetal Thymus Organ Culture and In Vitro Thymocyte Dulling Assay.- Protocol for Diversion of a CD4+Response to theT-Helper 2 CellPathway.- Quantitating Presentation of MHC Class I-Restricted Antigens.- Identification of CD8+T-Cell-Stimulating Antigen Genes in cDNA Libraries.- Generation of Antigen-Specific, LacZ-Inducible T-Cell Hybrids.
Purification of 20S Proteasomes.- Use of Proteasome Inhibitors to Examine Processing of Antigens for Major Histocompatibility Complex Class I Presentation.- Tracing the Route Taken by Peptides and Major Histocompatibility Complex Class I Molecules in Presentation of Exogenous Antigens.- Functional Analysis of Antigen Processing and Major Histocompatibility Complex Class II-Restricted Presentation.- Quantitation of Major Histocompatibility Complex Antigen Endocytosis Using Capture ELISA.- In Vitro Generation of Functional Human and Murine Dendritic Cells.- Genetic Engineering of Dendritic Cells Using Retrovirus-Based Gene Transfer Techniques.- Use of Vaccinia Virus Expression Vectors to Investigate Antigen Processing and Presentation.- Sindbis Virus-Based Vectors for the Study of Class I Antigen Presentation In Vitro and In Vivo.- Precision Genotyping of Human Leukocyte Antigen-A, -B and -C Loci Via Direct DNA Sequencing.- Photolabeling the Transporter Associated with Antigen Processing.- Application of One-Dimensional Isoelectric Focusing to Separate Different Major Histocompatibility Complex Class I Alleles and Determine Their Allelic Interactions with Transporter Associated with Antigen Processing (TAP).- Analysis of Major Histocompatibility Complex Class I Interactions with Endoplasmic Reticulum Proteins.- Methods of Purification of Heat Shock Protein-Peptide Complexes for Use as Vaccines Against Cancers and Infectious Diseases.- Random Peptide Libraries.- Neural Network Method for Predicting Peptides that Bind Major Histocompatibility Complex Molecules.- Analysis of T-Cell Response Using Altered Peptide Ligands.- Assays of Thymic Selection Fetal Thymus Organ Culture and In Vitro Thymocyte Dulling Assay.- Protocol for Diversion of a CD4+Response to theT-Helper 2 CellPathway.- Quantitating Presentation of MHC Class I-Restricted Antigens.- Identification of CD8+T-Cell-Stimulating Antigen Genes in cDNA Libraries.- Generation of Antigen-Specific, LacZ-Inducible T-Cell Hybrids.
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