title: A rapid culture technique produces functional dendritic-like cells from human acute myeloid leukemia cell lines.
abstract: Most anti-cancer immunotherapeutic strategies involving dendritic cells (DC) as vaccines rely upon the adoptive transfer of DC loaded with exogenous tumour-peptides. This study utilized human acute myeloid leukemia (AML) cells as progenitors from which functional dendritic-like antigen presenting cells (DLC) were generated, that constitutively express tumour antigens for recognition by CD8(+) T cells. DLC were generated from AML cell lines KG-1 and MUTZ-3 using rapid culture techniques and appropriate cytokines. DLC were evaluated for their cell-surface phenotype, antigen uptake and ability to stimulate allogeneic responder cell proliferation, and production of IFN-γ; compared with DC derived from normal human PBMC donors. KG-1 and MUTZ-3 DLC increased expression of CD80, CD83, CD86, and HLA-DR, and MUTZ-3 DLC downregulated CD14 and expressed CD1a. Importantly, both KG-1 and MUTZ-3-derived DLC promoted proliferation of allogeneic responder cells more efficiently than unmodified cells; neither cells incorporated FITC-labeled dextran, but both stimulated IFN-γ production from responding allogeneic CD8(+) T cells. Control DC produced from PBMC using the FastDC culture also expressed high levels of critical cell surface ligands and demonstrated good APC function. This paper indicates that functional DLC can be cultured from the AML cell lines KG-1 and MUTZ-3, and FastDC culture generates functional KG-1 DLC.
authors.affiliation: Immunotherapy Laboratory, Stem Cell R&D, Bristol Institute for Transfusion Sciences, NHS Blood and Transplant, North Bristol Park, Northway, Filton, Bristol BS34 7QH, UK.
authors.forename: Jian
authors.initials: J
authors.lastname: Ning
authors.forename: David
authors.initials: D
authors.lastname: Morgan
authors.forename: Derwood
authors.initials: D
authors.lastname: Pamphilon
mesh_terms: D000704:Analysis of Variance; D015703:Antigens, CD / Q000378:metabolism; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology / Q000378:metabolism; D018929:Cell Culture Techniques / Q000379:methods*; D045744:Cell Line, Tumor; D016207:Cytokines / Q000378:metabolism; D003713:Dendritic Cells / Q000276:immunology / Q000378:metabolism*; D003911:Dextrans / Q000737:chemistry; D005434:Flow Cytometry; D016650:Fluorescein-5-isothiocyanate / Q000031:analogs & derivatives / Q000737:chemistry; D015395:Histocompatibility Antigens Class I / Q000378:metabolism; D006801:Humans; D007371:Interferon-gamma / Q000378:metabolism; D015470:Leukemia, Myeloid, Acute / Q000276:immunology* / Q000473:pathology*; D007963:Leukocytes, Mononuclear / Q000276:immunology / Q000473:pathology; D010641:Phenotype
chemical_list: D015703:Antigens, CD; D016207:Cytokines; D003911:Dextrans; D015395:Histocompatibility Antigens Class I; C015219:fluorescein isothiocyanate dextran; D007371:Interferon-gamma; D016650:Fluorescein-5-isothiocyanate