title: Rapid generation of antigen-presenting cells from leukaemic blasts in acute myeloid leukaemia.
abstract: The ability of acute myeloid leukaemia (AML) cells to acquire dendritic cell (DC)-like characteristics in vitro with a rapid culture method based either on the phorbol ester PMA or calcium ionophores has been studied in comparison to conventional AML-DC cultures with the cytokines granulocyte-macrophage colony-stimulating factor (GM-CSF), tumour necrosis factor-alpha (TNF-alpha), interleukin-3 (IL-3), SCF, FLT3-L and IL-4. In all AML patients, antigen-presenting cells (APC) could be generated from leukaemic cells in 2 days by incubation with PMA or calcium ionophore (A23187 or ionomycin) in the presence as well as in the absence of IL-4. In 30 out of 36 patients APC could be generated after 2 weeks of culture in cytokine-enriched medium. AML-APC cultured with PMA or calcium ionophores immunophenotypically and functionally were at a more mature stage than those cultured in cytokine-enriched medium. The most mature APC were generated by calcium ionophore A23187 plus IL-4, as evidenced by the higher expression of CD40, CD80, CD86 and HLA-DR. Autologous T cell mediated cytotoxicity towards AML blast cells in vitro was observed in 2 cases tested. The persistence of cytogenetic abnormalities confirmed the leukaemic origin of the AML-APC. The generation of AML-APC was possible from freshly isolated as well as cryopreserved material. Our data show that generation of sufficient AML-APC by A23187 plus IL-4 is feasible, for vaccination purposes, in approximately 70% of AML specimens, offering a time-saving and cost-effective approach in preparing anti-leukaemia vaccines.
authors.affiliation: Department of Haematology, VU University Medical Centre, De Boelelaan 1117, Amsterdam, 1081 HV, The Netherlands.
authors.forename: Theresia M
authors.initials: TM
authors.lastname: Westers
authors.forename: Anita G M
authors.initials: AG
authors.lastname: Stam
authors.forename: Rik J
authors.initials: RJ
authors.lastname: Scheper
authors.forename: Johanna C
authors.initials: JC
authors.lastname: Regelink
authors.forename: Aggie W M
authors.initials: AW
authors.lastname: Nieuwint
authors.forename: Gerrit Jan
authors.initials: GJ
authors.lastname: Schuurhuis
authors.forename: Arjan A
authors.initials: AA
authors.lastname: van de Loosdrecht
authors.forename: Gert J
authors.initials: GJ
authors.lastname: Ossenkoppele
mesh_terms: D000208:Acute Disease; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000938:Antigen-Presenting Cells / Q000166:cytology* / Q000276:immunology; D015703:Antigens, CD / Q000096:biosynthesis; D000951:Antigens, Neoplasm / Q000032:analysis; D001854:Bone Marrow Cells / Q000166:cytology / Q000187:drug effects / Q000276:immunology; D000001:Calcimycin / Q000494:pharmacology*; D002118:Calcium / Q000502:physiology; D019496:Cancer Vaccines; D018929:Cell Culture Techniques / Q000379:methods*; D015925:Cryopreservation; D003602:Cytotoxicity, Immunologic; D005240:Feasibility Studies; D005260:Female; D006684:HLA-DR Antigens / Q000096:biosynthesis; D006801:Humans; D015847:Interleukin-4 / Q000494:pharmacology*; D007476:Ionophores / Q000494:pharmacology; D007951:Leukemia, Myeloid / Q000276:immunology* / Q000473:pathology / Q000628:therapy; D007959:Lymphocyte Culture Test, Mixed; D008297:Male; D008875:Middle Aged; D009360:Neoplastic Cells, Circulating / Q000187:drug effects / Q000276:immunology; D014411:Neoplastic Stem Cells / Q000166:cytology / Q000187:drug effects* / Q000276:immunology; D015203:Reproducibility of Results; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology*; D013997:Time Factors; D014021:Tissue Preservation / Q000379:methods; D014407:Tumor Cells, Cultured / Q000166:cytology / Q000187:drug effects / Q000276:immunology; D014611:Vaccination
chemical_list: D015703:Antigens, CD; D000951:Antigens, Neoplasm; D019496:Cancer Vaccines; D006684:HLA-DR Antigens; D007476:Ionophores; D015847:Interleukin-4; D000001:Calcimycin; D013755:Tetradecanoylphorbol Acetate; D002118:Calcium