Selective generation of different dendritic cell precursors from CD34+ cells by interleukin-6 and interleukin-3.
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-alpha). 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-alpha, 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-alpha, allowing the identification of developmental stages of DC2. Like true plasmacytoid DCs, these cells secreted interferon-alpha after TLR9-specific stimulation with a specific CpG nucleotide.
Encabo
Araceli
A
Instituto de Biología Celular, Agencia Valenciana de Ciencia y Tecnología, Avda del Cid 65 A, 46014 Valencia, Spain.
Solves
Pilar
P
Mateu
Emilia
E
Sepúlveda
Pilar
P
Carbonell-Uberos
Francisco
F
Miñana
María Dolores
MD
D018949:Antigens, CD1 / Q000276:immunology; D018952:Antigens, CD34 / Q000276:immunology*; D018826:CD13 Antigens / Q000276:immunology; D015815:Cell Adhesion Molecules / Q000276:immunology; D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation / Q000187:drug effects / Q000276:immunology; D002455:Cell Division / Q000187:drug effects / Q000276:immunology; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D036101:Cord Blood Stem Cell Transplantation / Q000379:methods*; D003713:Dendritic Cells / Q000166:cytology / Q000187:drug effects / Q000276:immunology*; D005260:Female; D005312:Fetal Blood / Q000166:cytology / Q000187:drug effects / Q000276:immunology*; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000187:drug effects / Q000276:immunology*; D006801:Humans; D007231:Infant, Newborn; D016898:Interferon-alpha / Q000187:drug effects / Q000378:metabolism; D007377:Interleukin-3 / Q000276:immunology / Q000494:pharmacology; D015850:Interleukin-6 / Q000276:immunology / Q000494:pharmacology; D007378:Interleukins / Q000276:immunology* / Q000494:pharmacology; D037181:Lectins, C-Type / Q000276:immunology; D018950:Lipopolysaccharide Receptors / Q000276:immunology; D008562:Membrane Glycoproteins / Q000819:agonists / Q000276:immunology; D010950:Plasma Cells / Q000166:cytology / Q000187:drug effects / Q000276:immunology; D011956:Receptors, Cell Surface / Q000819:agonists / Q000276:immunology; D051217:Toll-Like Receptor 9; D051193:Toll-Like Receptors; D014409:Tumor Necrosis Factor-alpha / Q000276:immunology / Q000494:pharmacology; D000099215:DC-Specific ICAM-3 Grabbing Nonintegrin
D018949:Antigens, CD1; D018952:Antigens, CD34; D018826:CD13 Antigens; D015815:Cell Adhesion Molecules; D016898:Interferon-alpha; D007377:Interleukin-3; D015850:Interleukin-6; D007378:Interleukins; D037181:Lectins, C-Type; D018950:Lipopolysaccharide Receptors; D008562:Membrane Glycoproteins; D011956:Receptors, Cell Surface; D051217:Toll-Like Receptor 9; D051193:Toll-Like Receptors; D014409:Tumor Necrosis Factor-alpha; D000099215:DC-Specific ICAM-3 Grabbing Nonintegrin; C419393:CD1a antigen; C495371:TLR9 protein, human