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Tetramer-guided epitope mapping: rapid identification and characterization of immunodominant CD4+ T cell epitopes from complex antigens.
Title | Tetramer-guided epitope mapping: rapid identification and characterization of immunodominant CD4+ T cell epitopes from complex antigens. |
Publication Type | Journal Article |
Year of Publication | 2001 |
Authors | Novak, EJ, Liu, AW, Gebe, JA, Falk, BA, Nepom, GT, Koelle, DM, Kwok, WW |
Journal | J Immunol |
Volume | 166 |
Issue | 11 |
Pagination | 6665-70 |
Date Published | 2001 Jun 1 |
ISSN | 0022-1767 |
Keywords | CD4-Positive T-Lymphocytes, Cell Line, Transformed, Clone Cells, Epitope Mapping, Epitopes, T-Lymphocyte, Herpes Simplex Virus Protein Vmw65, Herpesvirus 2, Human, HLA-DR alpha-Chains, HLA-DR Antigens, HLA-DRB1 Chains, Humans, Immunodominant Epitopes, Lymphocyte Activation, Macromolecular Substances, Peptide Fragments, Protein Binding |
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. |
Alternate Journal | J. Immunol. |
PubMed ID | 11359821 |