Generation of dendritic cells from bone marrow progenitors using GM‐CSF, TNF‐α, and additional cytokines: antagonistic effects of IL‐4 and IFN‐γ and selective involvement of TNF‐α receptor‐1

• Published in: Immunology 1997-08, Vol.91 (4), p.553-559
• Main Authors: LARDON, F., SNOECK, H.‐W., BERNEMAN, Z. N., VAN TENDELOO, V. F. I., NIJS, G., LENJOU, M., HENCKAERTS, E., BOECKXTAENS, C. J., VANDENABEELE, P., KESTENS, L. L., VAN BOCKSTAELE, D. R., VANHAM, G. L. E. E.
• Format: Article
• Language: English
• Subjects: Antigen Presentation; Antigens, CD34 - analysis; Cell Culture Techniques; Cell Differentiation - immunology; Cytokines - immunology; Dendritic Cells - immunology; Granulocyte-Macrophage Colony-Stimulating Factor - immunology; Hematopoietic Stem Cells - cytology; Hematopoietic Stem Cells - immunology; Humans; Immunophenotyping; Interleukin-4 - immunology; Mutation; Receptors, Tumor Necrosis Factor - immunology; Recombinant Proteins - immunology; Tumor Necrosis Factor-alpha - genetics; Tumor Necrosis Factor-alpha - immunology
• ISSN: 0019-2805; 1365-2567
• DOI: 10.1046/j.1365-2567.1997.00295.x

We report the generation of dendritic cells (DC) starting from CD34+ bone marrow (BM) progenitor cells, using a two‐stage culture system in which, besides granulocyte–macrophage colony‐stimulating factor (GM‐CSF) and tumour necrosis factor‐α (TNF‐α), stem‐cell factor (SCF) was added during the first 5 days, while interleukin‐4 (IL‐4) and/or interferon‐γ (IFN‐γ) were added during the secondary culture period of 9 days. Addition of IL‐4 favoured the outgrowth of CD1a+, HLA‐DR+, CD4+, CD40+, CD80+ but CD14− cells with dendritic morphology and strong antigen‐presenting capacity. Addition of IFN‐γ selectively induced HLA‐DR and CD86 but did not up‐regulate CD1a expression or antigen‐presenting capacity of the differentiated cells. An antagonism between IL‐4 and IFN‐γ could further be confirmed in that, as compared with IL‐4 alone, the simultaneous addition of IL‐4 and IFN‐γ to GM‐CSF plus TNF‐α during maturation reduced both the phenotypical (CD1a, CD4, CD40) and functional characteristics of DC. Using receptor‐specific TNF‐α mutants, we investigated the relative involvement of TNF‐α receptors R1 and R2 in the generation of DC. The induction of CD1a and HLA‐DR, as well as the increase in allostimulatory capacity were dependent on TNF‐R1 triggering, whereas triggering through TNF‐R2 had no measurable effect. We conclude first, that the expansion of DC from BM progenitors could most effectively be enhanced in a two‐stage culture assay using SCF, GM‐CSF, TNF‐α and IL‐4; second, that the effect of TNF‐α in DC generation involves signalling via the TNF‐R1 receptor; and third, that IFN‐γ counteracts some of the effects of IL‐4 in DC generation.