Tetramer-guided epitope mapping: rapid identification and characterization of immunodominant CD4+ T cell epitopes from complex antigens.
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.
Novak
E J
EJ
Virginia Mason Research Center, Benaroya Research Institute, Seattle, WA 98101, USA.
Liu
A W
AW
Gebe
J A
JA
Falk
B A
BA
Nepom
G T
GT
Koelle
D M
DM
Kwok
W W
WW
D015496:CD4-Positive T-Lymphocytes / Q000737:chemistry* / Q000378:metabolism / Q000821:virology; D002461:Cell Line, Transformed; D002999:Clone Cells / Q000737:chemistry / Q000276:immunology; D018604:Epitope Mapping / Q000379:methods*; D018984:Epitopes, T-Lymphocyte / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D006684:HLA-DR Antigens / Q000032:analysis / Q000096:biosynthesis / Q000378:metabolism; D059805:HLA-DR alpha-Chains; D059811:HLA-DRB1 Chains; D017405:Herpes Simplex Virus Protein Vmw65 / Q000032:analysis / Q000096:biosynthesis / Q000276:immunology / Q000378:metabolism; D018258:Herpesvirus 2, Human / Q000276:immunology; D006801:Humans; D016056:Immunodominant Epitopes / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D008213:Lymphocyte Activation; D046911:Macromolecular Substances; D010446:Peptide Fragments / Q000096:biosynthesis / Q000276:immunology / Q000378:metabolism; D011485:Protein Binding / Q000276:immunology
D018984:Epitopes, T-Lymphocyte; D006684:HLA-DR Antigens; D059805:HLA-DR alpha-Chains; D059811:HLA-DRB1 Chains; C553505:HLA-DRB1*04:01 antigen; D017405:Herpes Simplex Virus Protein Vmw65; D016056:Immunodominant Epitopes; D046911:Macromolecular Substances; D010446:Peptide Fragments