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.
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.
Knights
Ashley John
AJ
University of Tübingen, Section for Transplantation Immunology and Immunohaematology, Second Department of Internal Medicine, Zentrum für Medizinische Forschung ZMF, Waldhörnlestrasse 22, 72072 Tübingen, Germany.
Zaniou
Angeliki
A
Rees
Robert C
RC
Pawelec
Graham
G
Müller
Ludmila
L
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
D000939:Epitopes; D006684:HLA-DR Antigens; D010455:Peptides; D025721:WT1 Proteins; D011069:Poly I; D011066:Poly C; D007371:Interferon-gamma