Abstract
T cell responses to Ags involve recognition of selected peptide epitopes contained within the antigenic protein. In this report, we describe a new approach for direct identification of CD4+ T cell epitopes of complex Ags that uses human class II tetramers to identify reactive cells. With a panel of 60 overlapping peptides covering the entire sequence of the VP16 protein, a major Ag for HSV-2, we generated a panel of class II MHC tetramers loaded with peptide pools that were used to stain peripheral lymphocytes of an HSV-2 infected individual. With this approach, we identified four new DRA1*0101/DRB1*0401- and two DRA1*0101/DRB1*0404-restricted, VP16-specific epitopes. By using tetramers to sort individual cells, we easily obtained a large number of clones specific to these epitopes. Although DRA1*0101/DRB1*0401 and DRA1*0101/DRB1*0404 are structurally very similar, nonoverlapping VP16 epitopes were identified, illustrating high selectivity of individual allele polymorphisms within common MHC variants. This rapid approach to detecting CD4+ T cell epitopes from complex Ags can be applied to any known Ag that gives a T cell response.
Publication types
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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CD4-Positive T-Lymphocytes / chemistry*
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CD4-Positive T-Lymphocytes / metabolism
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CD4-Positive T-Lymphocytes / virology
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Cell Line, Transformed
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Clone Cells / chemistry
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Clone Cells / immunology
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Epitope Mapping / methods*
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Epitopes, T-Lymphocyte / analysis*
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Epitopes, T-Lymphocyte / chemistry
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Epitopes, T-Lymphocyte / metabolism
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HLA-DR Antigens / analysis
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HLA-DR Antigens / biosynthesis
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HLA-DR Antigens / metabolism
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HLA-DR alpha-Chains
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HLA-DRB1 Chains
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Herpes Simplex Virus Protein Vmw65 / analysis
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Herpes Simplex Virus Protein Vmw65 / biosynthesis
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Herpes Simplex Virus Protein Vmw65 / immunology
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Herpes Simplex Virus Protein Vmw65 / metabolism
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Herpesvirus 2, Human / immunology
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Humans
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Immunodominant Epitopes / analysis*
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Immunodominant Epitopes / chemistry
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Immunodominant Epitopes / metabolism
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Lymphocyte Activation
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Macromolecular Substances
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Peptide Fragments / biosynthesis
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Peptide Fragments / immunology
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Peptide Fragments / metabolism
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Protein Binding / immunology
Substances
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Epitopes, T-Lymphocyte
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HLA-DR Antigens
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HLA-DR alpha-Chains
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HLA-DRB1 Chains
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HLA-DRB1*04:01 antigen
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Herpes Simplex Virus Protein Vmw65
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Immunodominant Epitopes
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Macromolecular Substances
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Peptide Fragments