title: The alternative open reading frame of LAGE-1 gives rise to multiple promiscuous HLA-DR-restricted epitopes recognized by T-helper 1-type tumor-reactive CD4+ T cells.
abstract: The NY-ESO-1 and LAGE-1 genes are expressed by many human cancers, but not by normal tissues, with the exception of testis and placenta. The NY-ESO-1 and LAGE-1 genes give rise to multiple MHC class I and class II-presented epitopes derived from the open reading frames (ORF) 1 and 2. Here, we have investigated whether NY-ESO-1/LAGE-1 ORF2 encodes promiscuous MHC class II-restricted epitopes. Using a set of overlapping peptides from the ORF2 protein sequence and autologous dendritic cells (DCs) from normal donors and melanoma patients, we have identified three HLA-DRB1*0401-restricted peptide sequences from the LAGE-1 ORF2 that are capable of stimulating T-helper 1-type melanoma-reactive CD4+ T cells. From these bulk CD4+ T cells, we have generated CD4+ T-cell clones able to recognize not only peptide-pulsed DCs but also autologous DCs loaded with the LAGE-1 ORF2 protein. We have demonstrated that these peptides not only bind to multiple HLA-DR molecules apart from HLA-DRB1*0401 but also stimulate CD4+ T cells when presented in the context of these HLA-DR molecules. Furthermore, our binding data have delineated two additional sequences capable of broadly binding to multiple HLA-DR molecules. Altogether, these data support the immunogenicity of NY-ESO-1/LAGE-1 ORF2 gene products and clearly demonstrate their capability to stimulate T-helper 1 type CD4+ T cells. Because of the role of these cells in promoting long-lasting antitumor CTL responses, our data provide a rationale for cancer vaccine trials with peptides derived from the NY-ESO-1/LAGE-1 ORF2 for a large fraction of patients with NY-ESO-1/LAGE-1(+) tumors.
authors.affiliation: Department of Medicine and Melanoma Center, University of Pittsburgh School of Medicine, Pittsburgh, PA 15213, USA.
authors.forename: Maja
authors.initials: M
authors.lastname: Mandic
authors.forename: Christine
authors.initials: C
authors.lastname: Almunia
authors.forename: Stephanie
authors.initials: S
authors.lastname: Vicel
authors.forename: Daniel
authors.initials: D
authors.lastname: Gillet
authors.forename: Bratislav
authors.initials: B
authors.lastname: Janjic
authors.forename: Kara
authors.initials: K
authors.lastname: Coval
authors.forename: Bernard
authors.initials: B
authors.lastname: Maillere
authors.forename: John M
authors.initials: JM
authors.lastname: Kirkwood
authors.forename: Hassane M
authors.initials: HM
authors.lastname: Zarour
mesh_terms: D000595:Amino Acid Sequence; D000951:Antigens, Neoplasm / Q000235:genetics / Q000276:immunology; D000954:Antigens, Surface; D001665:Binding Sites; D045744:Cell Line, Tumor; D003713:Dendritic Cells / Q000276:immunology; D018984:Epitopes, T-Lymphocyte / Q000235:genetics / Q000276:immunology*; D006684:HLA-DR Antigens / Q000235:genetics / Q000276:immunology*; D059811:HLA-DRB1 Chains; D006801:Humans; D008545:Melanoma / Q000235:genetics / Q000276:immunology; D008565:Membrane Proteins; D008969:Molecular Sequence Data; D016366:Open Reading Frames / Q000276:immunology; D011506:Proteins / Q000235:genetics* / Q000276:immunology; D018417:Th1 Cells / Q000276:immunology*
chemical_list: D000951:Antigens, Neoplasm; D000954:Antigens, Surface; C104948:CTAG1B protein, human; C510162:CTAG2 protein, human; D018984:Epitopes, T-Lymphocyte; D006684:HLA-DR Antigens; D059811:HLA-DRB1 Chains; C553505:HLA-DRB1*04:01 antigen; D008565:Membrane Proteins; D011506:Proteins