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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
Author(s) -
Motta Maria R.,
Castellani Samantha,
Rizzi Simonetta,
Curti Antonio,
Gubinelli Francesco,
Fogli Miriam,
Ferri Elisa,
Cellini Claudia,
Baccarani Michele,
Lemoli Roberto M.
Publication year - 2003
Publication title -
british journal of haematology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.907
H-Index - 186
eISSN - 1365-2141
pISSN - 0007-1048
DOI - 10.1046/j.1365-2141.2003.04270.x
Subject(s) - cd14 , granulocyte macrophage colony stimulating factor , immunology , dendritic cell , leukapheresis , medicine , tumor necrosis factor alpha , cytokine , cd40 , andrology , chemistry , microbiology and biotechnology , biology , in vitro , immune system , stem cell , cytotoxic t cell , biochemistry , cd34
Summary. 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‐α (TNF‐α) or a cocktail of IL‐1β, IL‐6, TNF‐α and prostaglandin‐E 2 . 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 × 10 8 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‐α 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‐α‐2b was at least as efficient as IL‐4 in inducing the differentiation of mature, functional DC from monocytes.