title: [Identification of a novel HLA-A2-restrictive CTL epitope of an ovary cancer-associated antigen OVA66].
abstract: OBJECTIVE
  To identify a novel HLA-A2-restrictive CTL epitope of an ovary cancer-associated antigen OVA66.
  
  
  METHODS
  Dendritic cells (DCs), induced from peripheral blood mononuclear cells(PBMCs) by cytokines, were confirmed by morphological observation and FACS. Mature DCs were pulsed with each of the two synthesized peptides which were selected as possible CTL epitopes by software analysis. The pulsed DCs were used to stimulate autologous CD8+ T cells from an HLA-A2+ healthy donor. One week later, the peptides-pulsed autologous PBMCs were used to stimulate the CD8+ T cells for another 3 times at weekly intervals. The stimulated CD8+ T cells were used as CTLs. The cytotoxicity of CTLs to target cells was detected by lactate dehydrogenase (LDH) release assay and the number of T cells secreting antigen-specific IFN-gamma in CTLs was analyzed by enzyme-linked immunospot assay (ELISPOT).
  
  
  RESULTS
  The results of morphology observation and FACS indicated that mature DCs were induced from PBMCs. Of the two peptides, peptide L235(FLPDHINIV) induced peptide-specific CD8+ T cells that lysed HLA-A2+ T2 cells pulsed with L235 and OVA66+/HLA-A2+ SW480 cells. Compared with control peptide, L235 increased the number of IFN-gamma producing T cells.
  
  
  CONCLUSIONS
  This novel OVA66-derived CTL epitope L235 can induce HLA-A2-restrictive CTL response, which lays the foundation for preparation of tumor-specific peptide vaccine.
authors.affiliation: Shanghai Institute of Immunology, Shanghai Second Medical University, Shanghai 200025, China. jinshushu@sina.com
authors.forename: Shu
authors.initials: S
authors.lastname: Jin
authors.forename: Ying
authors.initials: Y
authors.lastname: Wang
authors.forename: Shu-jun
authors.initials: SJ
authors.lastname: Wang
authors.forename: Hui-zhen
authors.initials: HZ
authors.lastname: Zhang
authors.forename: Mei-xing
authors.initials: MX
authors.lastname: Li
authors.forename: Ying
authors.initials: Y
authors.lastname: Chen
authors.forename: Hai-liang
authors.initials: HL
authors.lastname: Ge
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000951:Antigens, Neoplasm / Q000276:immunology*; D019496:Cancer Vaccines / Q000276:immunology; D049109:Cell Proliferation; D003713:Dendritic Cells / Q000276:immunology; D018984:Epitopes, T-Lymphocyte / Q000032:analysis* / Q000276:immunology*; D005260:Female; D005434:Flow Cytometry; D015789:HLA-A2 Antigen / Q000276:immunology*; D006801:Humans; D007371:Interferon-gamma / Q000378:metabolism; D051379:Mice; D010051:Ovarian Neoplasms / Q000276:immunology*; D010455:Peptides / Q000737:chemistry / Q000276:immunology; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology* / Q000378:metabolism / Q000473:pathology
chemical_list: D000951:Antigens, Neoplasm; D019496:Cancer Vaccines; D018984:Epitopes, T-Lymphocyte; D015789:HLA-A2 Antigen; D010455:Peptides; D007371:Interferon-gamma