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Selective Generation of Different Dendritic Cell Precursors from CD34⁺ Cells by Interleukin-6 and Interleukin-3

^a Instituto de Biología Celular,Agencia Valenciana de Ciencia y Tecnología,Valencia, Spain;
^b Centro de Transfusión de la Comunidad Valenciana,Valencia, Spain;
^c Fundación Hospital General Universitario,Valencia, Spain

Key Words. Interleukin-6 • Interleukin-3 • Myeloid dendritic cell precursors • Plasmacytoid dendritic cells

Correspondence: María Dolores Miñana, Ph.D., Instituto de Biología Celular, Agencia Valenciana de Ciencia y Tecnología, Avda del Cid 65 A, 46014 Valencia, Spain. Telephone: 34-96-3868132; Fax: 34-96-3868109; e-mail: minyana_mdo{at}gva.es

There is a growing interest in generating dendritic cells (DCs) for using as vaccines. Several cytokines, especially stem cell factor (SCF) and FLT3-ligand (FL), have been identified as essential to produce large numbers of myeloid precursors and even to increase DC yield obtained by the action of granulocyte-macrophage colony-stimulating factor (GM-CSF) and tumor necrosis factor alpha (TNF-). However, there are few studies on the effect of the early-acting cytokines, commonly used to expand CD34⁺ progenitor cells, on DC generation. We report here that in the absence of serum, SCF, FL, and thrombopoietin (TPO) plus interleukin-6 (IL-6) and SCF, FL, and TPO plus IL-3 were able to generate CD14⁺CD1a^– and CD14^– CD1a⁺ myeloid DC precursors from CD34⁺ cells, but IL-6 had an inhibitory effect on the generation of CD14^– CD1a⁺ cells. Both DC precursors differentiated into mature DCs by GM-CSF, IL-4, and TNF-, and DCs obtained from both types of culture exhibited equal allostimulatory capacity. CD1a⁺ DCs generated could be identified on the basis of DC-specific intracellular adhesion molecule–grabbing nonintegrin (DC-SIGN) expression, a novel C-type lectin receptor expressed on dermal DCs but not on Langerhans cells. In addition, the inclusion of IL-3 to the culture medium induced the appearance of CD13^– cells that differentiated into plasmacytoid DC (DC2) on the addition of TNF-, allowing the identification of developmental stages of DC2. Like true plasmacytoid DCs, these cells secreted interferon- after TLR9-specific stimulation with a specific CpG nucleotide.

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