Abstract
The lack of high affinity reagents has made distinguishing T cells on the basis of antigen specificity difficult to accomplish. This unit provides protocols that utilize innovations in molecular design to permit construction of soluble multivalent MHC complexes (MHC-Ig dimers) with high avidity for cognate T cell receptors. MHC-Ig dimers display stable binding properties when they interact with antigen-specific T cells thus allowing their use in the staining of antigen-specific T cells by flow cytometry. Methods for constructing and detecting these MHC-Ig dimers are included along with protocols for applying their use for the quantitation of antigen-specific T cells.
MeSH terms
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Animals
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Antigens / immunology*
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Antigens, Surface / immunology
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CD8-Positive T-Lymphocytes / immunology*
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Cloning, Molecular
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DNA, Complementary / genetics
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DNA, Complementary / immunology
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Dimerization
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Enzyme-Linked Immunosorbent Assay / methods
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Flow Cytometry / methods
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Genetic Vectors / genetics
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Genetic Vectors / immunology
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Histocompatibility Antigens Class I / genetics
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Histocompatibility Antigens Class I / immunology*
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Histocompatibility Antigens Class II / genetics
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Histocompatibility Antigens Class II / immunology*
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Humans
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Immunoglobulins / genetics
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Immunoglobulins / immunology*
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Immunoglobulins / isolation & purification
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Polymerase Chain Reaction / methods
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Receptors, Antigen, T-Cell / immunology
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Recombinant Fusion Proteins / genetics
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Recombinant Fusion Proteins / immunology
Substances
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Antigens
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Antigens, Surface
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DNA, Complementary
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Histocompatibility Antigens Class I
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Histocompatibility Antigens Class II
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Immunoglobulins
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Receptors, Antigen, T-Cell
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Recombinant Fusion Proteins