Expansion of dendritic cell precursors from human CD34(+) progenitor cells isolated from healthy donor blood; growth factor combination determines proliferation rate and functional outcome.
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.
Bontkes
Hetty J
HJ
Department of Pathology, Vrije Universiteit Medical Center, Amsterdam, The Netherlands. hj.bontkes@vumc.nl
De Gruijl
Tanja D
TD
Schuurhuis
Gert Jan
GJ
Scheper
Rik J
RJ
Meijer
Chris J L M
CJ
Hooijberg
Erik
E
D000328:Adult; D017951:Antigen Presentation / Q000187:drug effects; D018949:Antigens, CD1 / Q000032:analysis; D000956:Antigens, Viral / Q000276:immunology; D001782:Blood Donors; D002455:Cell Division / Q000187:drug effects; D003713:Dendritic Cells / Q000166:cytology*; D004357:Drug Synergism; D016179:Granulocyte Colony-Stimulating Factor / Q000494:pharmacology; D016298:Hematopoietic Cell Growth Factors / Q000494:pharmacology*; D019650:Hematopoietic Stem Cell Mobilization; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000187:drug effects*; D006801:Humans; D007156:Immunologic Memory; D016130:Immunophenotyping; D016753:Interleukin-10 / Q000096:biosynthesis; D018664:Interleukin-12 / Q000096:biosynthesis; D007377:Interleukin-3 / Q000494:pharmacology; D015850:Interleukin-6 / Q000494:pharmacology; D007959:Lymphocyte Culture Test, Mixed; D008565:Membrane Proteins / Q000494:pharmacology; D019089:Stem Cell Factor / Q000494:pharmacology; D016176:T-Lymphocyte Subsets / Q000276:immunology; D013926:Thrombopoietin / Q000494:pharmacology; D014763:Viral Matrix Proteins / Q000276:immunology
D018949:Antigens, CD1; D000956:Antigens, Viral; C419393:CD1a antigen; D016298:Hematopoietic Cell Growth Factors; D007377:Interleukin-3; D015850:Interleukin-6; C031540:M-protein, influenza virus; D008565:Membrane Proteins; D019089:Stem Cell Factor; D014763:Viral Matrix Proteins; C084881:flt3 ligand protein; D016753:Interleukin-10; D016179:Granulocyte Colony-Stimulating Factor; D018664:Interleukin-12; D013926:Thrombopoietin