title: Cell lines transfected with the TAP inhibitor ICP47 allow testing peptide binding to a variety of HLA class I molecules.
abstract: The immediate early protein ICP47 of the Herpes simplex virus is known to block the human transporter associated with antigen processing (TAP), thereby creating a TAP-deficient phenotype in any human cell transfected with the corresponding cDNA. Exploiting this inhibitory activity, we constructed a selection of human cell lines each co-expressing one of the cDNAs of human leukocyte antigen (HLA) class I alleles HLA-A*1101, A24, A*3101, A*6601, B8 and B*1516, and the cDNA encoding the ICP47 molecule. The cell lines generated showed diminished HLA class I surface expression and the inhibition of the TAP function was confirmed in peptide translocation assays. The addition of specific exogenous peptide ligands restored the expression of the corresponding HLA class I molecules. Thus, the ICP47 transfectants provide us with a tool to closely examine peptide-HLA class I interactions, to confirm HLA class I ligand motifs and to test peptides predicted to bind.
authors.affiliation: Department of Immunology, Institute for Cell Biology, Eberhard-Karls University Tübingen, Germany.
authors.forename: J
authors.initials: J
authors.lastname: Gatfield
authors.forename: E
authors.initials: E
authors.lastname: Lammert
authors.forename: P
authors.initials: P
authors.lastname: Nickolaus
authors.forename: C
authors.initials: C
authors.lastname: Münz
authors.forename: S
authors.initials: S
authors.lastname: Rothenfusser
authors.forename: P
authors.initials: P
authors.lastname: Fisch
authors.forename: S
authors.initials: S
authors.lastname: Stevanović
authors.forename: H
authors.initials: H
authors.lastname: Schild
authors.forename: H G
authors.initials: HG
authors.lastname: Rammensee
authors.forename: D
authors.initials: D
authors.lastname: Arnold
mesh_terms: D000071181:ATP Binding Cassette Transporter, Subfamily B, Member 2; D018528:ATP-Binding Cassette Transporters / Q000037:antagonists & inhibitors*; D000483:Alleles; D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D005260:Female; D005434:Flow Cytometry; D015234:HLA-A Antigens / Q000378:metabolism; D015235:HLA-B Antigens / Q000378:metabolism; D018258:Herpesvirus 2, Human; D015395:Histocompatibility Antigens Class I / Q000378:metabolism*; D006801:Humans; D017874:Immediate-Early Proteins / Q000235:genetics / Q000378:metabolism*; D008024:Ligands; D051379:Mice; D009842:Oligopeptides / Q000378:metabolism*; D016415:Sequence Alignment; D014162:Transfection; D014764:Viral Proteins
chemical_list: D000071181:ATP Binding Cassette Transporter, Subfamily B, Member 2; D018528:ATP-Binding Cassette Transporters; D015234:HLA-A Antigens; D015235:HLA-B Antigens; D015395:Histocompatibility Antigens Class I; C105843:ICP47 protein, Herpes simplex virus; D017874:Immediate-Early Proteins; D008024:Ligands; D009842:Oligopeptides; C073119:TAP1 protein, human; C486171:Tap1 protein, mouse; D014764:Viral Proteins