title: Generation of dendritic cells from CD14+ monocytes positively selected by immunomagnetic adsorption for multiple myeloma patients enrolled in a clinical trial of anti-idiotype vaccination.
abstract: Circulating monocytes from multiple myeloma patients enrolled in a clinical study of anti-idiotype vaccination were labelled with clinical-grade anti-CD14 microbeads and positively selected with the CliniMACS instrument. Cells were then grown, according to good manufacturing practice guidelines, in fetal-calf-serum-free medium in cell culture bags and differentiated to dendritic cells (DC) with granulocyte-macrophage colony stimulating factor plus interleukin 4 (IL-4), followed by either tumour necrosis factor-alpha (TNF-alpha) or a cocktail of IL-1beta, IL-6, TNF-alpha and prostaglandin-E2. The CD14+ cell yield was increased from 17.6 +/- 6.5% to 93.8 +/- 6.3% (recovery 64.4 +/- 15.4%, viability > 97%). After cell culture, phenotypic analysis showed that 86.7 +/- 6.8% of the cells were DC: 2.27 +/- 0.9 x 108 DC/leukapheresis were obtained, which represented 20.7 +/- 4.6% of the initial number of CD14+ cells. Notably, the cytokine cocktail induced a significantly higher percentage and yield (28.6 +/- 3% of initial CD14+ cells) of DC than TNF-alpha alone, with secretion of larger amounts of IL-12, potent stimulatory activity on allogeneic T cells and efficient presentation of tumour idiotype to autologous T cells. Storage in liquid nitrogen did not modify the phenotype or functional characteristics of preloaded DC. The recovery of thawed, viable DC was 78 +/- 10%. Finally, interferon-alpha-2b was at least as efficient as IL-4 in inducing the differentiation of mature, functional DC from monocytes.
authors.affiliation: Institute of Haematology and Medical Oncology, L. & A. Seràgnoli, University of Bologna, Bologna, Italy.
authors.forename: Maria R
authors.initials: MR
authors.lastname: Motta
authors.forename: Samantha
authors.initials: S
authors.lastname: Castellani
authors.forename: Simonetta
authors.initials: S
authors.lastname: Rizzi
authors.forename: Antonio
authors.initials: A
authors.lastname: Curti
authors.forename: Francesco
authors.initials: F
authors.lastname: Gubinelli
authors.forename: Miriam
authors.initials: M
authors.lastname: Fogli
authors.forename: Elisa
authors.initials: E
authors.lastname: Ferri
authors.forename: Claudia
authors.initials: C
authors.lastname: Cellini
authors.forename: Michele
authors.initials: M
authors.lastname: Baccarani
authors.forename: Roberto M
authors.initials: RM
authors.lastname: Lemoli
mesh_terms: D000888:Antibodies, Anti-Idiotypic / Q000008:administration & dosage*; D002454:Cell Differentiation; D002478:Cells, Cultured; D015925:Cryopreservation; D003713:Dendritic Cells / Q000276:immunology*; D006801:Humans; D018189:Immunomagnetic Separation; D016130:Immunophenotyping; D016219:Immunotherapy, Adoptive / Q000379:methods*; D000077190:Interferon alpha-2; D016898:Interferon-alpha / Q000494:pharmacology; D015847:Interleukin-4 / Q000494:pharmacology; D007963:Leukocytes, Mononuclear / Q000166:cytology / Q000276:immunology*; D018950:Lipopolysaccharide Receptors / Q000276:immunology; D009101:Multiple Myeloma / Q000628:therapy*; D011994:Recombinant Proteins
chemical_list: D000888:Antibodies, Anti-Idiotypic; D000077190:Interferon alpha-2; D016898:Interferon-alpha; D018950:Lipopolysaccharide Receptors; D011994:Recombinant Proteins; D015847:Interleukin-4