Abstract
The recognition of defined antigen-MHC complexes by antigen-specific T cells forms the molecular basis of T cell immunity. It has been shown that fluorescently labeled recombinant MHC tetramers can be utilized to detect antigen-specific T cells by flow cytometry. Since this first description, MHC tetramers and other types of MHC multimers have become a core tool to monitor the development of disease- and therapy-induced antigen-specific T cell responses both in humans and in animal model systems. This unit describes a set of protocols that transform classical MHC multimer technology into a high-throughput platform, allowing one to produce large collections of MHC class I molecules charged with different peptides. This technology is based on the development of conditional MHC ligands that can be triggered to self-destruct while in the MHC-bound state.
MeSH terms
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Animals
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Biotinylation / methods
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Carbon-Nitrogen Ligases / biosynthesis
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Carbon-Nitrogen Ligases / isolation & purification
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Enzyme-Linked Immunosorbent Assay / methods
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Escherichia coli Proteins / biosynthesis
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Escherichia coli Proteins / isolation & purification
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Fluorescent Dyes / chemistry
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Histocompatibility Antigens Class I / biosynthesis
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Histocompatibility Antigens Class I / chemistry*
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Histocompatibility Antigens Class I / immunology*
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Humans
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Indicators and Reagents / chemistry
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Ligands
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Mice
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Peptides / chemistry*
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Peptides / immunology*
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Peptides / radiation effects
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Protein Multimerization
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Protein Renaturation
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Recombinant Proteins / biosynthesis
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Recombinant Proteins / chemistry
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Recombinant Proteins / isolation & purification
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Repressor Proteins / biosynthesis
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Repressor Proteins / isolation & purification
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Ultraviolet Rays
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beta 2-Microglobulin / biosynthesis
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beta 2-Microglobulin / chemistry
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beta 2-Microglobulin / immunology
Substances
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Escherichia coli Proteins
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Fluorescent Dyes
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Histocompatibility Antigens Class I
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Indicators and Reagents
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Ligands
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Peptides
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Recombinant Proteins
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Repressor Proteins
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beta 2-Microglobulin
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Carbon-Nitrogen Ligases
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birA protein, E coli