Cell lines transfected with the TAP inhibitor ICP47 allow testing peptide binding to a variety of HLA class I molecules.
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.
Gatfield
J
J
Department of Immunology, Institute for Cell Biology, Eberhard-Karls University Tübingen, Germany.
Lammert
E
E
Nickolaus
P
P
Münz
C
C
Rothenfusser
S
S
Fisch
P
P
Stevanović
S
S
Schild
H
H
Rammensee
H G
HG
Arnold
D
D
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
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