title: Large-scale production of human tumorcytotoxic macrophages grown from blood monocytes of cancer patients.
abstract: For adoptive immunotherapy protocols using cells of the macrophage (M phi) system, well differentiated and functionally competent effector cells are required. In this presentation the generation of a large number of M phi grown in vitro from blood monocytes (mo) is reported. Mononuclear cells (MNC) were collected by cytapheresis and subsequent Ficoll centrifugation. Mean yield was 6.9 x 10(9) MNC (range from 3 x 10(9) to 1.2 x 10(10), n = 18) with a mean mo count of 22 +/- 14%. MNC were cultured at 5 x 10(6)/ml in suspension on hydrophobic Teflon foils with 2% autologous serum for 7 days with recombinant human interferon-gamma (rhIFN-gamma) being present for the last 18 h of culture. Cells were harvested and activated mo-derived M phi separated from lymphocytes by counter-current centrifugal elutriation. On average, 42% of mo cultured could be recovered as M phi, the maximal number of M phi generated being 1.7 x 10(9) with a purity of up to 96%. Mo-derived M phi appeared to be mature by their expression of maturation-associated antigens and proved to be cytotoxic to allogeneic tumor targets in vitro. They secreted large quantities of tumor necrosis factor alpha (TNF-alpha), interleukin-6 (IL-6), and granulocyte-macrophage colony stimulating factor (GM-CSF) upon stimulation with endotoxin. Using the technology described, this study revealed that large amounts of tumorcytotoxic M phi can be generated from the peripheral blood of cancer patients to be used in adoptive immunotherapy trials.
authors.affiliation: Medizinische Klinik der Albert-Ludwigs-Universität, Freiburg, Germany.
authors.forename: W
authors.initials: W
authors.lastname: Brugger
authors.forename: C
authors.initials: C
authors.lastname: Scheibenbogen
authors.forename: S
authors.initials: S
authors.lastname: Krause
authors.forename: R
authors.initials: R
authors.lastname: Andreesen
mesh_terms: D000328:Adult; D000943:Antigens, Differentiation / Q000032:analysis; D002452:Cell Count; D002454:Cell Differentiation; D002478:Cells, Cultured; D046508:Culture Techniques / Q000295:instrumentation / Q000379:methods*; D016238:Cytapheresis; D003601:Cytotoxicity Tests, Immunologic; D004731:Endotoxins / Q000494:pharmacology; D005260:Female; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000378:metabolism; D006801:Humans; D016219:Immunotherapy, Adoptive; D007371:Interferon-gamma / Q000494:pharmacology; D015850:Interleukin-6 / Q000378:metabolism; D008262:Macrophage Activation; D008264:Macrophages / Q000187:drug effects / Q000276:immunology / Q000378:metabolism; D008297:Male; D008875:Middle Aged; D009000:Monocytes; D009369:Neoplasms / Q000097:blood / Q000276:immunology / Q000473:pathology* / Q000628:therapy; D011994:Recombinant Proteins; D014182:Transplantation, Autologous; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism
chemical_list: D000943:Antigens, Differentiation; D004731:Endotoxins; D015850:Interleukin-6; D011994:Recombinant Proteins; D014409:Tumor Necrosis Factor-alpha; D007371:Interferon-gamma; D016178:Granulocyte-Macrophage Colony-Stimulating Factor