Abstract
We report in this work the expression and characterization of class I molecules expressed as single-chain trimers consisting of an antigenic peptide-spacer-beta(2)-microglobulin-spacer H chain. Our results indicate that these single-chain constructs assemble efficiently, maintain their covalent structure, and are unusually stable at the cell surface. Consequently, these constructs are at least 1000-fold less accessible to exogenous peptide than class I molecules loaded with endogenous peptides, and they are potent simulators of peptide-specific CTL and Abs. Our combined findings suggest that single-chain trimers may have applications as DNA vaccines against virus infection or tumors.
Publication types
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Animals
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B-Lymphocytes / immunology*
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Egg Proteins / chemistry
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Egg Proteins / genetics
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Egg Proteins / immunology
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Egg Proteins / physiology
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Epitopes, B-Lymphocyte / immunology
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Epitopes, T-Lymphocyte / immunology
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Genetic Vectors / chemical synthesis
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Genetic Vectors / immunology
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Genetic Vectors / physiology
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H-2 Antigens / chemistry
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H-2 Antigens / genetics
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H-2 Antigens / immunology
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H-2 Antigens / physiology
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Histocompatibility Antigens Class I / chemistry*
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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 I / physiology*
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L Cells
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Lymphocyte Activation / genetics
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Lymphocyte Activation / immunology*
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Mice
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Mice, Inbred BALB C
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Mice, Inbred C3H
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Mice, Inbred C57BL
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Mice, Transgenic
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Ovalbumin / chemistry
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Ovalbumin / genetics
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Ovalbumin / immunology
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Ovalbumin / physiology
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Peptide Fragments / chemical synthesis*
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Peptide Fragments / genetics
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Peptide Fragments / immunology
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Peptide Fragments / physiology*
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Structure-Activity Relationship
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T-Lymphocytes / immunology*
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beta 2-Microglobulin / chemical synthesis
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beta 2-Microglobulin / genetics
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beta 2-Microglobulin / physiology
Substances
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Egg Proteins
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Epitopes, B-Lymphocyte
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Epitopes, T-Lymphocyte
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H-2 Antigens
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H-2Kb protein, mouse
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Histocompatibility Antigens Class I
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OVA-8
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Peptide Fragments
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beta 2-Microglobulin
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Ovalbumin