Progenitor recruitment and in vitro expansion of immunostimulatory dendritic cells from human CD34+ bone marrow cells by c-kit-ligand, GM-CSF, and TNF alpha.
Several cytokines have been identified that support the development of dendritic cells from murine and human precursor populations, most notably GM-CSF, TNF alpha, and IL-4. We have been interested in human bone marrow as a source of defined CD34+ progenitors to generate large numbers of autologous dendritic cells for use as adjuvants in immune based therapy. In serum-replete conditions with c-kit-ligand, GM-CSF, and TNF alpha, dendritic cells constitute approximately 10-15% of the myeloid progeny (equivalent to approximately 1.7 x 10(6) dendritic cells per single ml of starting bone marrow); and they develop together with granulocytic intermediates and monocytes in the same cultures. CD14- dendritic cells share expression of class II MHC and costimulatory ligands with CD14+ monocyte progeny, but only the CD14- HLA-DR+ dendritic cells are highly stimulatory of resting unprimed T cells. We have further identified a novel colony that develops in the presence of GM-CSF and TNF alpha alongside typical CFU-GM, which is comprised of dendritic cells mixed with < or = 15% monocytes (CFU-DC/mono). c-kit-ligand recruits and expands early progenitors responsive to the dendritic cell-differentiating effects of GM-CSF and TNF alpha, effecting a 100- to 1000-fold greater expansion of CFU-DC/mono by 14d and 21d respectively than does the combination of GM-CSF and TNF alpha without c-kit-ligand.(ABSTRACT TRUNCATED AT 250 WORDS)
Laboratory of Cellular Physiology and Immunology, Rockefeller University, New York, NY, USA.
P
P
Szabolcs
E D
ED
Feller
M A
MA
Moore
J W
JW
Young
D018952:Antigens, CD34 / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects; D003114:Colony-Forming Units Assay; D003713:Dendritic Cells / Q000166:cytology / Q000187:drug effects* / Q000276:immunology; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000008:administration & dosage*; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000187:drug effects* / Q000276:immunology; D006801:Humans; D066298:In Vitro Techniques; D009000:Monocytes / Q000166:cytology; D019089:Stem Cell Factor / Q000008:administration & dosage*; D014409:Tumor Necrosis Factor-alpha / Q000008:administration & dosage*
D018952:Antigens, CD34; D019089:Stem Cell Factor; D014409:Tumor Necrosis Factor-alpha; D016178:Granulocyte-Macrophage Colony-Stimulating Factor