title: Pocket 4 of the HLA-DR(alpha,beta 1*0401) molecule is a major determinant of T cells recognition of peptide.
abstract: To investigate the functional roles of individual HLA-DR residues in T cell recognition, transfectants expressing wild-type or mutant DR(alpha,beta 1*0401) molecules with single amino acid substitutions at 14 polymorphic positions of the DR beta 1*0401 chain or 19 positions of the DR alpha chain were used as antigen-presenting cells for five T cell clones specific for the influenza hemagglutinin peptide, HA307-19. Of the six polymorphic positions in the DR beta floor that were examined, mutations at only two positions eliminated T cell recognition: positions 13 (four clones) and 28 (one clone). In contrast, individual mutations at DR beta positions 70, 71, 78, and 86 on the alpha helix eliminated recognition by each of the clones, and mutations at positions 74 and 67 eliminated recognition by four and two clones, respectively. Most of the DR alpha mutations had minimal or no effect on most of the clones, although one clone was very sensitive to changes in the DR alpha chain, with loss of recognition in response to 10 mutants. Mutants that abrogated recognition by all of the clones were assessed for peptide binding, and only the beta 86 mutation drastically decreased peptide binding. Single amino acid substitutions at polymorphic positions in the central part of the DR beta alpha helix disrupted T cell recognition much more frequently than substitutions in the floor, suggesting that DR beta residues on the alpha helix make relatively greater contributions than those in the floor to the ability of the DR(alpha,beta 1*0401) molecule to present HA307-19. The data indicate that DR beta residues 13, 70, 71, 74, and 78, which are located in pocket 4 of the peptide binding site in the crystal structure of the DR1 molecule, exert a major and disproportionate influence on the outcome of T cell recognition, compared with other polymorphic residues.
authors.affiliation: Department of Immunology, G. D. Searle & Co., St. Louis, Missouri 63198.
authors.forename: X T
authors.initials: XT
authors.lastname: Fu
authors.forename: C P
authors.initials: CP
authors.lastname: Bono
authors.forename: S L
authors.initials: SL
authors.lastname: Woulfe
authors.forename: C
authors.initials: C
authors.lastname: Swearingen
authors.forename: N L
authors.initials: NL
authors.lastname: Summers
authors.forename: F
authors.initials: F
authors.lastname: Sinigaglia
authors.forename: A
authors.initials: A
authors.lastname: Sette
authors.forename: B D
authors.initials: BD
authors.lastname: Schwartz
authors.forename: R W
authors.initials: RW
authors.lastname: Karr
mesh_terms: D000595:Amino Acid Sequence; D002460:Cell Line; D006684:HLA-DR Antigens / Q000737:chemistry / Q000276:immunology*; D006801:Humans; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D017433:Protein Structure, Secondary; D013329:Structure-Activity Relationship; D013601:T-Lymphocytes / Q000276:immunology*
chemical_list: D006684:HLA-DR Antigens