title: In vitro induction of human cytotoxic T lymphocyte responses against peptides of mutant and wild-type p53.
abstract: The central role of the p53 tumor suppressor gene product in oncogenesis is gradually being clarified. Point mutations in the p53 tumor suppressor gene are common in most human cancers and are often associated with p53 protein overexpression. Overexpressed wild-type or mutant determinants of the p53 protein thus represent an attractive target for immunotherapy of cancer directed against a structure involved in malignant transformation. An important step towards this goal is identification of epitopes of p53 that can be recognized by human cytotoxic T lymphocytes. We identified peptides of (mutant) p53 capable of binding to HLA-A2.1 in an in vitro assay. These HLA-A2.1-binding peptides were utilized for in vitro induction of primary cytotoxic T lymphocyte responses using a human processing-defective cell line (174CEM.T2) as antigen-presenting cell. These cells display "empty" HLA class I surface molecules, that can efficiently be loaded with a single peptide. We obtained CD8+ cytotoxic T lymphocyte clones capable of specifically lysing target cells loaded with wild-type or tumor-specific mutant p53 peptides. This strategy allows the in vitro initiation of human cytotoxic T lymphocyte responses against target molecules of choice.
authors.affiliation: Department of Immunohaematology and Blood Bank, University Hospital, Leiden, The Netherlands.
authors.forename: J G
authors.initials: JG
authors.lastname: Houbiers
authors.forename: H W
authors.initials: HW
authors.lastname: Nijman
authors.forename: S H
authors.initials: SH
authors.lastname: van der Burg
authors.forename: J W
authors.initials: JW
authors.lastname: Drijfhout
authors.forename: P
authors.initials: P
authors.lastname: Kenemans
authors.forename: C J
authors.initials: CJ
authors.lastname: van de Velde
authors.forename: A
authors.initials: A
authors.lastname: Brand
authors.forename: F
authors.initials: F
authors.lastname: Momburg
authors.forename: W M
authors.initials: WM
authors.lastname: Kast
authors.forename: C J
authors.initials: CJ
authors.lastname: Melief
mesh_terms: D000595:Amino Acid Sequence; D002460:Cell Line; D003602:Cytotoxicity, Immunologic; D015789:HLA-A2 Antigen / Q000378:metabolism; D006801:Humans; D008969:Molecular Sequence Data; D009154:Mutation; D013329:Structure-Activity Relationship; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*; D016159:Tumor Suppressor Protein p53 / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism
chemical_list: D015789:HLA-A2 Antigen; D016159:Tumor Suppressor Protein p53