Lentiviral transduction of immune cells

Swainson, L.; Mongellaz, C.; Adjali, O.; Vicente, R.; Taylor, N.

Methods Mol Biol

2008 / vol 415 / pages 301-20


Gene transfer into mammalian cells has been of crucial importance for studies determining the role of specific genes in the differentiation and cell fate of various hematopoietic lineages. Until recently, the majority of these studies were performed in transformed cell lines due to difficulties in achieving levels of transfection of greater than 1-3% in primary hematopoietic cells. Vectors based on retrovirus and lentivirus backbones have revolutionized our ability to transfer genes into primary hematopoietic cells. These vectors have allowed extensive ex vivo and in vivo studies following introduction of a gene of interest and have been used clinically in individuals suffering from cancers, infections, and genetic diseases. Ex vivo lentiviral gene transfer can result in efficient transduction of progenitor cells (>80%) that can then be further differentiated into immune lineage cells including T, B, dendritic, or natural killer cells. Alternatively, differentiated immune cells can themselves be transduced ex vivo with lentiviral vectors. Here, we discuss optimization of technologies for human immunodeficiency virus (HIV)-based gene transfer into murine and human progenitor and immune cell lineages.

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Animals; Cells, Cultured; Mice; Transgenes; Transfection; B-Lymphocytes/virology; Centrifugation; Dendritic Cells/virology; Genetic Vectors/genetics; Hematopoietic Stem Cells/virology; Lentivirus/*genetics; Leukocytes, Mononuclear/*virology; Plasmids/genetics; Promoter Regions (Genetics)/genetics; T-Lymphocytes/virology; Titrimetry; Transduction, Genetic/*methods; Virion/metabolism

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