title: Development and characterization of HAT-sensitive Ewing tumour cells for immunotherapy.
abstract: BACKGROUND
  Despite improvements in the treatment of patients with Ewing family tumours (EFT) during the past decades, the prognosis for patients with advanced disease is still unsatisfying. New treatment strategies have to be developed.
  
  
  METHODS
  A hypoxanthine/aminopterin/thymidine (HAT)-sensitive EFT cell line was developed by repetitive treatment of the EFT cell line SK-N-MC with 8'-azaguanine (8AG). By using DNA microarrays, the gene expression profile of this cell line was characterized. Immunostimulatory activity was assessed by mixed lymphocyte/tumour cell culture (MLTC). Artificial fusion of tumour cells and dendritic cells was visualized by flow cytometry.
  
  
  RESULTS
  After selection of 8AG-resistant cells, a cell line with high sensitivity for treatment with HAT was obtained. Expression of the X chromosome inactivation specific transcript XIST was higher in HAT-sensitive cells. Nevertheless, HAT-sensitive cells retained the EFT-associated gene expression profile. Moreover, in the presence of HAT, it was possible to use these cells without irradiation as stimulatory cells in MLTC or as fusion partner for dendritic cells.
  
  
  CONCLUSIONS
  HAT-sensitive EFT cells might be an interesting tool for the development of new immunotherapeutic approaches for the treatment of EFT.
authors.affiliation: Universitätsklinik und Poliklinik für Kinder- und Jugendmedizin, Klinikum der Martin-Luther-Universität Halle-Wittenberg, Ernst Grube Str. 40, D-06097 Halle, Germany.
authors.forename: Christian
authors.initials: C
authors.lastname: Pfeifle
authors.forename: Kerstin
authors.initials: K
authors.lastname: Reinhardt
authors.forename: Sigrid
authors.initials: S
authors.lastname: Heins
authors.forename: Stefan
authors.initials: S
authors.lastname: Burdach
authors.forename: Martin S
authors.initials: MS
authors.lastname: Staege
mesh_terms: D000630:Aminopterin / Q000494:pharmacology*; D000951:Antigens, Neoplasm / Q000096:biosynthesis / Q000276:immunology; D001375:Azaguanine / Q000494:pharmacology; D002148:Calmodulin-Binding Proteins / Q000096:biosynthesis / Q000276:immunology; D019496:Cancer Vaccines / Q000276:immunology; D045744:Cell Line, Tumor; D020869:Gene Expression Profiling; D015789:HLA-A2 Antigen / Q000276:immunology / Q000378:metabolism; D006801:Humans; D019271:Hypoxanthine / Q000494:pharmacology*; D016233:Immunotherapy, Active / Q000379:methods*; D020411:Oligonucleotide Array Sequence Analysis; D010455:Peptides / Q000276:immunology; D051789:Proto-Oncogene Protein c-fli-1 / Q000096:biosynthesis / Q000276:immunology; D034802:RNA-Binding Protein EWS; D016601:RNA-Binding Proteins / Q000096:biosynthesis / Q000276:immunology; D012512:Sarcoma, Ewing / Q000235:genetics / Q000276:immunology* / Q000473:pathology; D013936:Thymidine / Q000494:pharmacology*
chemical_list: D000951:Antigens, Neoplasm; D002148:Calmodulin-Binding Proteins; D019496:Cancer Vaccines; C500986:EWSR1 protein, human; C078433:FLI1 protein, human; D015789:HLA-A2 Antigen; D010455:Peptides; D051789:Proto-Oncogene Protein c-fli-1; D034802:RNA-Binding Protein EWS; D016601:RNA-Binding Proteins; D019271:Hypoxanthine; D000630:Aminopterin; D001375:Azaguanine; D013936:Thymidine