Proliferation in monocyte-derived dendritic cell cultures is caused by progenitor cells capable of myeloid differentiation

Dendritic cells (DC) can be generated by culture of adherent peripheral blood (PB) cells in the presence of granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin-4 (IL-4). There is controversy as to whether these DC arise from proliferating precursors or simply from differentiati...

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Bibliographic Details
Published in:Blood 1998-09, Vol.92 (5), p.1598-1607
Main Authors: CAVANAGH, L. L, SAAL, R. J, GRIMMETT, K. L, THOMAS, R
Format: Article
Language:English
Subjects:
Antigen-Presenting Cells - cytology
Biological and medical sciences
Cell Differentiation
Cell Division
Cells, Cultured
Culture Media
Dendritic Cells - cytology
Flow Cytometry
Fundamental and applied biological sciences. Psychology
Fundamental immunology
Granulocyte-Macrophage Colony-Stimulating Factor - pharmacology
Hematopoietic Stem Cells - cytology
Humans
Immunobiology
Immunophenotyping
Interleukin-4 - pharmacology
Ki-67 Antigen - analysis
Lipopolysaccharides - pharmacology
Monocytes - cytology
Monocytes, macrophages
Myeloid cells: ontogeny, maturation, markers, receptors
Peroxidase - analysis
Tumor Necrosis Factor-alpha - pharmacology
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Summary:Dendritic cells (DC) can be generated by culture of adherent peripheral blood (PB) cells in the presence of granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin-4 (IL-4). There is controversy as to whether these DC arise from proliferating precursors or simply from differentiation of monocytes. DC were generated from myeloid-enriched PB non-T cells or sorted monocytes. DC generated from either population functioned as potent antigen-presenting cells. Uptake of [3H]-thymidine was observed in DC cultured from myeloid-enriched non-T cells. Addition of lipopolysaccharide or tumor necrosis factor-alpha led to maturation of the DC, but did not inhibit proliferation. Ki67(+) cells were observed in cytospins of these DC, and by double staining were CD3(-)CD19(-)CD11c-CD40(-) and myeloperoxidase+, suggesting that they were myeloid progenitor cells. Analysis of the starting population by flow cytometry demonstrated small numbers of CD34(+)CD33(-)CD14(-) progenitor cells, and numerous granulocyte-macrophage colony-forming units were generated in standard assays. Thus, production of DC in vitro from adherent PB cells also enriches for progenitor cells that are capable of proliferation after exposure to GM-CSF. Of clinical importance, the yield of DC derived in the presence of GM-CSF and IL-4 cannot be expanded beyond the number of starting monocytes.
ISSN:0006-4971
1528-0020
DOI:10.1182/blood.v92.5.1598