Differentiation of human dendritic cells from monocytes in vitro using granulocyte-macrophage colony stimulating factor and low concentration of interleukin-4

• Published in: Vojnosanitetski pregled Vol. 60; no. 5; pp. 531 - 538
• Main Authors: Colić, Miodrag, Jandrić, Dusan, Stojić-Vukanić, Zorica, Antić-Stanković, Jelena, Popović, Petar, Vasilijić, Sasa, Milosavljević, Petar, Balint, Bela
• Format: Journal Article
• Language: English
• Published: Serbia Military Health Department, Ministry of Defance, Serbia 01-09-2003
• Subjects: Apoptosis; Cell Adhesion; Cell Differentiation - drug effects; Cells, Cultured; dendritic cells; Dendritic Cells - cytology; Dendritic Cells - drug effects; Dendritic Cells - immunology; granulocyte-macrophage colony-stimulating factor; Granulocyte-Macrophage Colony-Stimulating Factor - pharmacology; Humans; Immunophenotyping; interleukin-4; Interleukin-4 - pharmacology; Lymphocyte Activation; monocytes; Monocytes - cytology; Monocytes - drug effects; phenotype; T-lymphocytes
• DOI: 10.2298/VSP0305531C
• EISSN: 2406-0720
• ISSN: 0042-8450

Several laboratories have developed culture systems that allow the generation of large numbers of human dendritic cells (DC) from monocytes using granulocyte-macrophage colony stimulating factor (GM-CSF), and interleukin-4 (IL-4). In this work we provided evidence that GM-CSF (100 ng/ml) in combination with a low concentration of IL-4 (5 ng/ml) was efficient in the generation of immature, non-adherent, monocyte-derived DC as the same concentration of GM-CSF, and ten times higher concentration of IL-4 (50 ng/ml). This conclusion was based on the similar phenotype profile of DC, such as the expression of CD1a, CD80, CD86, and HLA-DR, down-regulation of CD14, and the absence of CD83, as well as on their similar allostimulatory activity for T cells. A higher number of cells remained adherent in cultures with lower concentrations of IL-4 than in cultures with higher concentrations of the cytokine. However, most of these adherent cells down-regulated CD14 and stimulated the proliferation of alloreactive T cells. In contrast, adherent cells cultivated with GM-CSF alone were predominantly macrophages, as judged by the expression of CD14 and the inefficiency to stimulate alloreactive T cells. DC generated in the presence of lower concentrations of IL-4 had higher proapoptotic potential for the Jurkat cell line than DC differentiated with higher concentrations of IL-4, suggesting their stronger cytotoxic, anti-tumor effect.