Generation of peptide MHC class I monomers and multimers through ligand exchange.
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.
Toebes
Mireille
M
The Netherlands Cancer Institute, Amsterdam, The Netherlands.
Rodenko
Boris
B
The Netherlands Cancer Institute, Amsterdam, The Netherlands.
Ovaa
Huib
H
The Netherlands Cancer Institute, Amsterdam, The Netherlands.
Schumacher
Ton N M
TNM
The Netherlands Cancer Institute, Amsterdam, The Netherlands.
D000818:Animals; D019921:Biotinylation / Q000379:methods; D019731:Carbon-Nitrogen Ligases / Q000096:biosynthesis / Q000302:isolation & purification; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D029968:Escherichia coli Proteins / Q000096:biosynthesis / Q000302:isolation & purification; D005456:Fluorescent Dyes / Q000737:chemistry; D015395:Histocompatibility Antigens Class I / Q000096:biosynthesis / Q000737:chemistry* / Q000276:immunology*; D006801:Humans; D007202:Indicators and Reagents / Q000737:chemistry; D008024:Ligands; D051379:Mice; D010455:Peptides / Q000737:chemistry* / Q000276:immunology* / Q000528:radiation effects; D055503:Protein Multimerization; D020673:Protein Renaturation; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification; D012097:Repressor Proteins / Q000096:biosynthesis / Q000302:isolation & purification; D014466:Ultraviolet Rays; D001613:beta 2-Microglobulin / Q000096:biosynthesis / Q000737:chemistry / Q000276:immunology
D029968:Escherichia coli Proteins; D005456:Fluorescent Dyes; D015395:Histocompatibility Antigens Class I; D007202:Indicators and Reagents; D008024:Ligands; D010455:Peptides; D011994:Recombinant Proteins; D012097:Repressor Proteins; D001613:beta 2-Microglobulin; D019731:Carbon-Nitrogen Ligases; C058125:birA protein, E coli