Title Antigen processing : methods and protocols / edited by Peter van Endert

Published New York : Humana Press, ©2013

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Description 1 online resource (xvii, 589 pages) : color illustrations Series Methods in molecular biology, 1064-3745 ; 960 Methods in molecular biology (Clifton, N.J.) ; 960. 1064-3745 Contents Purification of large cytosolic proteases for in vitro assays: 20S and 26S proteasomes -- Analysis of proteasome generated antigenic peptides by mass spectrometry -- Using protease inhibitors in antigen presentation assays -- Kinetic studies of cytoplasmic antigen processing and production of MHC class I ligands -- Assaying peptide translocation by the peptide transporter TAP -- In vitro reconstitution of the MHC class I peptide-loading complex -- Studying mhc class i peptide loading and exchange in vitro -- Measuring synthesis and degradation of MHC class I molecules -- Studying ubiquitination of MHC class I molecules -- Evaluation of viral interference with MHC class I-restricted antigen processing and presentation using a flow cytometry-based approach -- Determining the activity of the transporter associated with antigen processing in the compartments of the secretory pathway -- Biochemical large-scale identification of MHC class I ligands -- Establishing MHC class I peptide motifs -- Quantitating MHC class I ligand production and presentation using TCR-like antibodies -- Biochemical analysis of naturally processed antigenic peptides presented by MHC class I molecules -- Identifying source proteins for MHC class I-presented peptides -- Purification, preparation, and use of chaperone-peptide complexes for tumor immunotherapy -- Recombinant poxviruses: Versatile tools for immunological assays -- Bioinformatics identification of antigenic peptide: Predicting the specificity of major MHC class I and II pathway players -- Evaluating CD8(+) T cell responses in vitro -- Cloning CD8(+) cytolytic T lymphocytes -- Production of CD4(+) and CD8 (+) T cell hybridomas -- Tracking antigen-specific CD8(+) T cells using MHC class I multimers -- The purification of large numbers of antigen presenting dendritic cells from mouse spleen -- Preparation of dendritic cells by in vitro cultures -- Monitoring dendritic cell activation and maturation -- Monitoring the intracellular routing of internalized antigens by immunofluorescence microscopy -- Isolation of a specialized, antigen-loaded early endosomal subpopulation by flow cytometry -- Preparing antigens suitable for cross-presentation assays in vitro and in vivo -- Gene transduction in human monocyte-derived dendritic cells using lentiviral vectors -- Pulse-chase analysis for studies of MHC class II biosynthesis, maturation, and peptide loading -- Assembly of matched [alpha/beta] subunits to HLA class II peptide receptors -- Studying MHC class II peptide loading and editing in vitro -- Study of the allogeneic response induced by endothelial cells expressing hla class ii after lentiviral transduction -- Antigen processing for mhc presentation via macroautophagy -- Studying MHC class II transport in dendritic cells -- In vitro digestion with proteases producing MHC class II ligands -- MHC-II ubiquitination -- Studying MHC class II presentation of immobilized antigen by B lymphocytes -- Production of primary human CD4(+) T cell lines and clones -- Analyzing antigen recognition by natural killer T cells -- Chromatofocusing purification of CD1B-antigen complexes and their analysis by isoelectric focusing Summary Antigen processing is a biological process that prepares antigens for the presentation to special cells in the immune system called T lymphocytes. In Antigen Processing: Methods and Protocols, expert researchers in the field provide a comprehensive set of protocols for studying presentation of antigens produced in the standard processing pathways for MHC class I and class II molecules. The chapters follow chronology of intracellular processing events, ending with recognition of peptide-MHC complexes at the cell surface by T lymphocytes. Written in the highly successful Methods in Molecular Biology™ series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and key tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Antigen Processing: Methods and Protocols is designed for beginners and experts interested in studying antigen processing Analysis Medicine Immunology Monoclonal antibodies Biomedicine Antibodies biomedische wetenschappen immunologie antilichamen Medicine (General) Geneeskunde (algemeen) Bibliography Includes bibliographical references and index Notes English Subject Antigens -- Analysis -- Laboratory manuals Antigen presenting cells. Antigen Presentation Antigen presenting cells Antigens -- Analysis Genre/Form Laboratory manuals Laboratory manuals. Manuels de laboratoire. Form Electronic book Author Endert, Peter van ISBN 9781627032186 1627032185

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