title: Prediction of an HLA-DR-binding peptide derived from Wilms' tumour 1 protein and demonstration of in vitro immunogenicity of WT1(124-138)-pulsed dendritic cells generated according to an optimised protocol.
abstract: The Wilms' tumour 1 (WT1) protein is over-expressed in several types of cancer including leukaemias and might therefore constitute a novel target for immunotherapy. Recently, human leucocyte antigen (HLA) class I-binding WT1 peptides have been identified and shown to stimulate CD8(+) T cells in vitro. For maximal CD8 cell efficacy, CD4(+) helper T cells responding to major histocompatibility complex (MHC) class II-binding epitopes are required. Here, we report that scanning the WT1 protein sequence using an evidence-based predictive computer algorithm (SYFPEITHI) yielded a peptide WT1(124-138) predicted to bind the HLA-DRB1*0401 molecule with high affinity. Moreover, synthetic WT1(124-138)-peptide-pulsed dendritic cells (DC), generated according to a protocol optimised in the present study, sensitised T cells in vitro to proliferate and secrete interferon-gamma (IFN-gamma) when rechallenged with specific peptide-pulsed DC, but not with peripheral blood mononuclear cells (PBMC). These results suggest that the WT1 protein may yield epitopes immunogenic to CD4 as well as CD8 T cells, and therefore constitute a novel potential target for specific immunotherapy.
authors.affiliation: University of Tübingen, Section for Transplantation Immunology and Immunohaematology, Second Department of Internal Medicine, Zentrum für Medizinische Forschung ZMF, Waldhörnlestrasse 22, 72072 Tübingen, Germany.
authors.forename: Ashley John
authors.initials: AJ
authors.lastname: Knights
authors.forename: Angeliki
authors.initials: A
authors.lastname: Zaniou
authors.forename: Robert C
authors.initials: RC
authors.lastname: Rees
authors.forename: Graham
authors.initials: G
authors.lastname: Pawelec
authors.forename: Ludmila
authors.initials: L
authors.lastname: Müller
mesh_terms: D000465:Algorithms; D015496:CD4-Positive T-Lymphocytes / Q000378:metabolism; D018414:CD8-Positive T-Lymphocytes / Q000378:metabolism; D002448:Cell Adhesion; D002455:Cell Division; D002469:Cell Separation; D003713:Dendritic Cells / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes; D005434:Flow Cytometry; D006684:HLA-DR Antigens / Q000737:chemistry*; D006801:Humans; D016130:Immunophenotyping; D007371:Interferon-gamma / Q000494:pharmacology; D007963:Leukocytes, Mononuclear / Q000378:metabolism; D009000:Monocytes / Q000378:metabolism; D010455:Peptides / Q000737:chemistry*; D011066:Poly C / Q000737:chemistry; D011069:Poly I / Q000737:chemistry; D011485:Protein Binding; D012984:Software; D025721:WT1 Proteins / Q000737:chemistry* / Q000378:metabolism
chemical_list: D000939:Epitopes; D006684:HLA-DR Antigens; D010455:Peptides; D025721:WT1 Proteins; D011069:Poly I; D011066:Poly C; D007371:Interferon-gamma