Expression of the nlsLacz gene in dendritic cells derived from retrovirally transduced peripheral blood CD34+ cells.
OBJECTIVE
Gene transfer and expression of exogenous genetic information coding for an immunogenic protein in antigen presenting cells (APCs) can promote an immune response. This was investigated by retroviral transfer of a marker gene into CD34+ derived APCs.


METHODS
To achieve long term expression of a specific transgene in APCs, G-CSF mobilized peripheral blood CD34+ cell populations were retrovirally transduced with the bacterial nlsLacZ, a marker gene used here as a model, in the presence of IL-3, IL-6, GM-CSF and SCF prior to being induced to differentiate into dendritic and macrophage cells by GM-CSF and TNF-a.


RESULTS
Addition of IL-4 was found to induce dendritic differentiation preferentially by inhibiting proliferation and differentiation of the macrophage lineage. As assessed by X-Gal staining, LacZ gene expression was observed in cells from both the dendritic lineage (CD1a+/CD14-) which still exhibits the highest immunostimulatory activity in mixed lymphocyte reaction and from the macrophage lineage (CD1a-/ CD14+).


CONCLUSIONS
This study sets out the possibility of transducing dendritic and macrophage progenitors present in the CD34+ cell population and in using a marker gene such as nlsLacZ to study gene expression in antigen presenting cell compartments.
Chischportich
C
C
Centre de Thérapie Génique, Institut Paoli-Calmettes, Marseille, France.
Bagnis
C
C
Galindo
R
R
Mannoni
P
P
D017951:Antigen Presentation; D018949:Antigens, CD1 / Q000032:analysis; D018952:Antigens, CD34 / Q000032:analysis; D002454:Cell Differentiation / Q000187:drug effects; D019070:Cell Lineage; D003713:Dendritic Cells / Q000201:enzymology* / Q000821:virology; D004790:Enzyme Induction; D017930:Genes, Reporter; D005822:Genetic Vectors / Q000235:genetics*; D016298:Hematopoietic Cell Growth Factors / Q000494:pharmacology; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism / Q000821:virology; D006801:Humans; D015847:Interleukin-4 / Q000494:pharmacology; D007763:Lac Operon; D018950:Lipopolysaccharide Receptors / Q000032:analysis; D007959:Lymphocyte Culture Test, Mixed; D008264:Macrophages / Q000187:drug effects / Q000201:enzymology / Q000821:virology; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics; D012190:Retroviridae / Q000235:genetics*; D014162:Transfection; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology; D001616:beta-Galactosidase / Q000096:biosynthesis* / Q000235:genetics
D018949:Antigens, CD1; D018952:Antigens, CD34; D016298:Hematopoietic Cell Growth Factors; D018950:Lipopolysaccharide Receptors; D011993:Recombinant Fusion Proteins; D014409:Tumor Necrosis Factor-alpha; D015847:Interleukin-4; D001616:beta-Galactosidase