Massive ex vivo generation of functional dendritic cells from mobilized CD34+ blood progenitors for anticancer therapy

S. Siena, M. Di Nicola, M. Bregni, R. Mortarini, A. Anichini, L. Lombardi, F. Ravagnani, G. Parmiani, A. M. Gianni

Research output: Contribution to journalArticlepeer-review

Abstract

We report that blood cell autografts, collected by single leukapheresis in cancer patients (n = 11) at the time of mobilization of hematopoietic progenitors into peripheral blood following anticancer therapy with high-dose cyclophosphamide (HD-CTX) plus interleukin-3 (IL-3) and granulocyte colony-stimulating factor (G-CSF/filgrastim), comprise 1.98 ± 0.39 x 105/kg (mean ± SE) CD34+ progenitors of dendritic cells (DCs). This number corresponds to 140-fold more progenitors than in a control autograft collected in the steady state. DCs derived from mobilized CD34+ cells, morphologically and immunophenotypically undistinguishable from skin Langerhans cells and DCs from bone marrow and cord blood CD34+ cells, are shown to be powerful stimulators of allogeneic T cell proliferation in primary MLR and of autologous HLA-DR-restricted CD4+ T cell proliferation in response to presentation of xenogenic antigens. We show that the GM-CSF-plus-TNF-α-dependent ex vivo generation of DCs from mobilized CD34+ cells is 2.5-fold enhanced by flk-2/flt-3 ligand or c-kit ligand (stem cell factor) and five-fold enhanced by a combination of these growth factors. In addition, the optimal serum for the generation of DCs is autologous HD-CTX recovery-phase serum rather than fetal calf serum (FCS) or steady-state human serum, which are clinically inadequate and ineffective, respectively. In practice, the stimulation of CD34+ cells in a blood cell autograft (15.75 ± 2.46 x 106/kg) provided by the above four growth factors should permit ex vivo generation of approximately 40 x 109 DCs in an adult patient. These new findings provide advantageous tools for the large-scale generation of DCs that are potentially usable for clinical protocols of immunotherapy or vaccination in patients undergoing cancer treatment.

Original languageEnglish
Pages (from-to)1463-1471
Number of pages9
JournalExperimental Hematology
Volume23
Issue number14
Publication statusPublished - 1995

Keywords

  • c-kit ligand
  • CD34
  • Dendritic cell progenitors
  • Dendritic cells
  • flk-2/flt-3 ligand
  • Peripheral blood hematopoietic progenitors

ASJC Scopus subject areas

  • Cancer Research
  • Cell Biology
  • Genetics
  • Hematology
  • Oncology
  • Transplantation

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