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Generation of dendritic cells from peripheral blood of patients at different stages of chronic myeloid leukemia

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Abstract

We report a method to generate dendritic cells (DC) from frozen leukapheresis products of patients with chronic myeloid leukemia (CML), using sterile culture bags and serum-free culture medium, ie conditions feasible for re-infusion into the patient as part of immunother-apeutic protocols. Leukapheresis products were stored from harvests performed either at diagnosis (13 patients) or after chemotherapy with subsequent granulocyte colony stimulating factor (G-CSF) administration (9 patients), for Peripheral Blood Stem Cell (PBSC) collections. In the presence of optimal concentrations of GM-CSF (50 ng/ml) and IL-4 (40 ng/ml) CML progenitors differentiated on day 7 and 14 of culture to DC, expressing CD1a, HLA-DRand CD86 surface antigens. Mature DCs exhibited on average 12-fold higher allo-stimulatory capacity for CD4+ and CD8+ cells compared to non-cultured PBMC in mixed lymphocyte reaction (MLR). Only DCs obtained from CML patients at diagnosis exhibited bcr/abl fusion gene when tested by fluorescentin situ hybridization (FISH). CD34-selection on leukapheresis products from diagnosis (7 patients) resulted in later maturation of DCs (after 14–15 d), compared to the non-selected PBMC. CD34-selection significantly increased the DC growth, and improved the allo-stimulatory capacity in MLR (on average on day 14, 3.5- and 2.3-fold, respectively). Large differences were observed between individual patients and different leukapheresis products from the same patient. Our report demonstrates the possibility to generateex vivo autologous functionally active DC in CML in a way that allows their clinical application as immunotherapeutic agents.

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Authors and Affiliations

  1. Department of Clinical Immunology and Transfusion Medicine, Immunohemotherapy Unit, Karolinska Hospital, S-171 76, Stockholm, Sweden

    C Zheng, O Stromberg & M Hansson

  2. Department of Internal Medicine, Division of Hematology and Infectious Diseases, Karolinska Hospital, S-171 76, Stockholm, Sweden

    P Pisa

  3. Department of Oncology, Radiumhemmet, CCK R8;01, Karolinska Hospital, S-171 76, Stockholm, Sweden

    P Pisa

  4. Department of Clinical Genetics, Karolinska Hospital, S-171 76, Stockholm, Sweden

    E Blennow

Authors
  1. C Zheng
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  2. P Pisa
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  3. O Stromberg
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  4. E Blennow
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  5. M Hansson
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Correspondence to P Pisa.

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Zheng, C., Pisa, P., Stromberg, O. et al. Generation of dendritic cells from peripheral blood of patients at different stages of chronic myeloid leukemia. Med Oncol 17, 270–278 (2000). https://doi.org/10.1007/BF02782191

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  • DOI: https://doi.org/10.1007/BF02782191

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