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*Substance via MeSH
The Journal of Immunology, 2001, 166: 6665-6670.
Copyright © 2001 by The American Association of Immunologists

Tetramer-Guided Epitope Mapping: Rapid Identification and Characterization of Immunodominant CD4+ T Cell Epitopes from Complex Antigens1

Erik J. Novak*,{dagger}, Andrew W. Liu*, John A. Gebe*, Ben A. Falk*, Gerald T. Nepom*, David M. Koelle{ddagger},§ and William W. Kwok2,*

* Virginia Mason Research Center, Benaroya Research Institute, Seattle, WA 98101; {dagger} R. H. Williams Laboratory, Molecular and Cellular Biology Program, and {ddagger} Departments of Medicine and Laboratory Medicine, University of Washington, Seattle, WA 98195; and § Fred Hutchinson Cancer Research Center, Seattle, WA 98109

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.




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