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

Maria R. Motta, Samantha Castellani, Simonetta Rizzi, Antonio Curti, Francesco Gubinelli, Miriam Fogli, Elisa Ferri, Claudia Cellini, Michele Baccarani, Roberto M. Lemoli

Research output: Contribution to journalArticlepeer-review

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-α (TNF-α) or a cocktail of IL-1β, IL-6, TNF-α 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 × 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-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.

Original languageEnglish
Pages (from-to)240-250
Number of pages11
JournalBritish Journal of Haematology
Volume121
Issue number2
DOIs
Publication statusPublished - Apr 2003

Keywords

  • CD14 cells
  • Cellular immunotherapy
  • Dendritic cells
  • Multiple myeloma

ASJC Scopus subject areas

  • Hematology

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