Expression of the nlsLacz gene in dendritic cells derived from retrovirally transduced peripheral blood CD34+ cells

Haematologica. 1999 Mar;84(3):195-203.

Abstract

Background and 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.

Design and 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+).

Interpretation and 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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antigen Presentation
  • Antigens, CD1 / analysis
  • Antigens, CD34 / analysis
  • Cell Differentiation / drug effects
  • Cell Lineage
  • Dendritic Cells / enzymology*
  • Dendritic Cells / virology
  • Enzyme Induction
  • Genes, Reporter
  • Genetic Vectors / genetics*
  • Hematopoietic Cell Growth Factors / pharmacology
  • Hematopoietic Stem Cells / cytology*
  • Hematopoietic Stem Cells / drug effects
  • Hematopoietic Stem Cells / metabolism
  • Hematopoietic Stem Cells / virology
  • Humans
  • Interleukin-4 / pharmacology
  • Lac Operon*
  • Lipopolysaccharide Receptors / analysis
  • Lymphocyte Culture Test, Mixed
  • Macrophages / drug effects
  • Macrophages / enzymology
  • Macrophages / virology
  • Recombinant Fusion Proteins / biosynthesis
  • Recombinant Fusion Proteins / genetics
  • Retroviridae / genetics*
  • Transfection
  • Tumor Necrosis Factor-alpha / pharmacology
  • beta-Galactosidase / biosynthesis*
  • beta-Galactosidase / genetics

Substances

  • Antigens, CD1
  • Antigens, CD34
  • Hematopoietic Cell Growth Factors
  • Lipopolysaccharide Receptors
  • Recombinant Fusion Proteins
  • Tumor Necrosis Factor-alpha
  • Interleukin-4
  • beta-Galactosidase