Dendritic cells (DCs) can be successfully generated from leukemic blasts in individual patients with AML or MDS: an evaluation of different methods

J Immunother. Feb-Mar 2010;33(2):185-99. doi: 10.1097/CJI.0b013e3181b8f4ce.

Abstract

Myeloid-leukemic cells (AML, MDS, CML) can be differentiated to leukemia-derived dendritic cell [DC (DCleu)] potentially presenting the whole leukemic antigen repertoire without knowledge of distinct leukemia antigens and are regarded as promising candidates for a vaccination strategy. We studied the capability of 6 serum-free DC culture methods, chosen according to different mechanisms, to induce DC differentiation in 137 cases of AML and 52 cases of MDS. DC-stimulating substances were cytokines ("standard-medium", "MCM-Mimic", "cytokine-method"), bacterial lysates ("Picibanil"), double-stranded RNA ["Poly (I:C)"] or a cytokine bypass method ("Ca-ionophore"). The quality/quantity of DC generated was estimated by flow cytometry studying (co) expressions of "DC"antigens, costimulatory, maturation, and blast-antigens. Comparing these methods on average 15% to 32% DC, depending on methods used, could be obtained from blast-containing mononuclear cells (MNC) in AML/MDS cases with a DC viability of more than 60%. In all, 39% to 64% of these DC were mature; 31% to 52% of leukemic blasts could be converted to DCleu and DCleu-proportions in the suspension were 2% to 70% (13%). Average results of all culture methods tested were comparable, however not every given case of AML could be differentiated to DC with 1 selected method. However performing a pre-analysis with 3 DC-generating methods (MCM-Mimic, Picibanil, Ca-ionophore) we could generate DC in any given case. Functional analyses provided proof, that DC primed T cells to antileukemia-directed cytotoxic cells, although an anti-leukemic reaction was not achieved in every case. In summary our data show that a successful, quantitative DC/DCleu generation is possible with the best of 3 previously tested methods in any given case. Reasons for different functional behaviors of DC-primed T cells must be evaluated to design a practicable DC-based vaccination strategy.

MeSH terms

  • Adult
  • Aged
  • Aged, 80 and over
  • Antigen Presentation / drug effects
  • Antigens, Differentiation / metabolism
  • Antigens, Neoplasm / immunology
  • Antigens, Neoplasm / metabolism
  • Cancer Vaccines*
  • Cell Culture Techniques / methods*
  • Cell Differentiation / drug effects
  • Cell Separation
  • Culture Media, Serum-Free
  • Cytokines / metabolism
  • Cytokines / pharmacology
  • Cytotoxicity, Immunologic
  • Dendritic Cells / drug effects
  • Dendritic Cells / immunology
  • Dendritic Cells / metabolism
  • Dendritic Cells / pathology*
  • Female
  • Flow Cytometry
  • Humans
  • Leukemia, Myeloid, Acute / immunology
  • Leukemia, Myeloid, Acute / pathology*
  • Leukemia, Myeloid, Acute / therapy
  • Male
  • Middle Aged
  • Myelodysplastic Syndromes / immunology
  • Myelodysplastic Syndromes / pathology*
  • Myelodysplastic Syndromes / therapy
  • Picibanil / pharmacology
  • Poly I-C / pharmacology
  • T-Lymphocytes, Cytotoxic / immunology

Substances

  • Antigens, Differentiation
  • Antigens, Neoplasm
  • Cancer Vaccines
  • Culture Media, Serum-Free
  • Cytokines
  • Picibanil
  • Poly I-C