Expansion of dendritic cell precursors from human CD34(+) progenitor cells isolated from healthy donor blood; growth factor combination determines proliferation rate and functional outcome

J Leukoc Biol. 2002 Aug;72(2):321-9.

Abstract

CD34(+) haematopoietic progenitor cells, which circulate at extremely low frequencies in peripheral blood, are used to generate dendritic cells (DC) in vitro. Here, we describe a method to grow large numbers of DC precursors from these low frequent cells. Different combinations of early acting haematopoietic growth factors supported expansion of CD34(+) cells. CD1a(+) DC derived from precursors, expanded in fms-like tyrosine kinase-3 ligand (Flt3-L), stem-cell factor (SCF), interleukin (IL)-3, and IL-6, were less potent antigen-presenting cells (APC) compared to CD1a(+) DC derived from precursors expanded in Flt3-L, trombopoietine (TPO), and SCF. Furthermore, the latter produced high levels of IL-12 and low levels of IL-10, a cytokine profile favorable for the priming cytotoxic T cells. In contrast, a mean increase of total cell number of 453-fold was obtained with Flt3-L, SCF, IL-3, and IL-6, and this increase was only 38-fold with Flt3-L, TPO, and SCF. Sequential cultures of both cocktails resulted in high numbers of potent APC, which can be useful DC-based cancer vaccines.

Publication types

  • Comparative Study

MeSH terms

  • Adult
  • Antigen Presentation / drug effects
  • Antigens, CD1 / analysis
  • Antigens, Viral / immunology
  • Blood Donors
  • Cell Division / drug effects
  • Dendritic Cells / cytology*
  • Drug Synergism
  • Granulocyte Colony-Stimulating Factor / pharmacology
  • Hematopoietic Cell Growth Factors / pharmacology*
  • Hematopoietic Stem Cell Mobilization
  • Hematopoietic Stem Cells / cytology
  • Hematopoietic Stem Cells / drug effects*
  • Humans
  • Immunologic Memory
  • Immunophenotyping
  • Interleukin-10 / biosynthesis
  • Interleukin-12 / biosynthesis
  • Interleukin-3 / pharmacology
  • Interleukin-6 / pharmacology
  • Lymphocyte Culture Test, Mixed
  • Membrane Proteins / pharmacology
  • Stem Cell Factor / pharmacology
  • T-Lymphocyte Subsets / immunology
  • Thrombopoietin / pharmacology
  • Viral Matrix Proteins / immunology

Substances

  • Antigens, CD1
  • Antigens, Viral
  • CD1a antigen
  • Hematopoietic Cell Growth Factors
  • Interleukin-3
  • Interleukin-6
  • M-protein, influenza virus
  • Membrane Proteins
  • Stem Cell Factor
  • Viral Matrix Proteins
  • flt3 ligand protein
  • Interleukin-10
  • Granulocyte Colony-Stimulating Factor
  • Interleukin-12
  • Thrombopoietin