Unilineage monocytopoiesis in hematopoietic progenitor culture: switching cytokine treatment at all Mo developmental stages induces differentiation into dendritic cells.
We have developed a new culture system whereby human hematopoietic progenitors purified from adult peripheral blood extensively proliferate and gradually differentiate into >95% pure monocytic (Mo) cells. At all developmental stages treatment with interleukin (IL)-4+granulocyte-macrophage colony-stimulating factor or IL-4+c-Kit-ligand+FLT-3 ligand switched the Mo precursors into dendritic cells (DCs). The switching capacity decreased only at the end of the culture, when most Mo cells matured to macrophages. Moreover, the Mo precursors were highly susceptible to transduction with lentiviral vectors: once switched to DCs, they maintained the transgene expression, as well as the phenotype and function of the DC lineage. Our results provide new insight into the potential role of the Mo lineage as a reservoir of DCs in vivo. Furthermore, the methodology for transduction of Mo precursors provides a tool to generate genetically modified, normally functioning DCs potentially useful for immunotherapy.
Montesoro
E
E
Department of Hematology, Oncology and Molecular Medicine, Istituto Superiore di Sanità, Rome, Italy. e.montes@iss.it
Castelli
G
G
Morsilli
O
O
Nisini
R
R
Stafsnes
M H
MH
Carè
A
A
Peschle
C
C
Chelucci
C
C
D019070:Cell Lineage; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D016207:Cytokines / Q000494:pharmacology*; D003713:Dendritic Cells / Q000166:cytology* / Q000502:physiology; D005786:Gene Expression Regulation; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000494:pharmacology; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000187:drug effects; D006801:Humans; D007167:Immunotherapy; D015847:Interleukin-4 / Q000494:pharmacology; D016086:Lentivirus / Q000235:genetics; D008565:Membrane Proteins / Q000494:pharmacology; D009000:Monocytes / Q000737:chemistry / Q000166:cytology* / Q000187:drug effects; D038042:Myelopoiesis / Q000187:drug effects*; D010641:Phenotype; D016187:Receptors, Granulocyte-Macrophage Colony-Stimulating Factor / Q000032:analysis / Q000235:genetics; D019089:Stem Cell Factor / Q000494:pharmacology; D014161:Transduction, Genetic; D019076:Transgenes
D016207:Cytokines; D008565:Membrane Proteins; D016187:Receptors, Granulocyte-Macrophage Colony-Stimulating Factor; D019089:Stem Cell Factor; C084881:flt3 ligand protein; D015847:Interleukin-4; D016178:Granulocyte-Macrophage Colony-Stimulating Factor