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- Title
Pre-clinical evaluation of an in vitro selection protocol for the enrichment of transduced CD34<sup>+</sup> cell-derived human dendritic cells.
- Authors
Evans, J T; Cravens, P; Gatlin, J; Kelly, P F; Lipsky, P E; Garcia, J V
- Abstract
The efficient genetic modification of CD34[sup +] cell-derived dendritic cells (DC) will provide a significant advancement towards the development of immunotherapy protocols for cancer, autoimmune disorders and infectious diseases. Recent reports have described the transduction of CD34[sup +] cells via retrovirus- and lentivirus-based gene transfer vectors and subsequent differentiation into functional DC. Since there is significant apprehension regarding the clinical uses of HIV-based vectors, in this report, we compare a murine leukemia virus (MLV)- and a human immunodeficiency virus (HIV)-based bicistronic vector for gene transfer into human CD34[sup +] cells and subsequent differentiation into mature DC. Each vector expressed both EGFP and the dominant selectable marker DHFR[sub L22Y] allowing for the enrichment of marked cells in the presence of the antifolate drug trimetrexate (TMTX). Both MLV-based and HIV-based vectors efficiently transduced cytokine mobilized human peripheral blood CD34[sup +] cells. However, in vitro expansion and differentiation in the presence of GM-CSF, TNF-α, Flt-3L, SCF and IL-4 resulted in a reduction in the percentage of DC expressing the transgene. Selection with TMTX during differentiation increased the percentage of marked DC, resulting in up to 79% (MLV vector) and up to 94% (lentivirus-vector) transduced cells expressing EGFP without loss of DC phenotype. Thus, MLV-based vectors and in vitro selection of transduced human DC show great promise for immunotherapy protocols.
- Subjects
DENDRITIC cells; GENETIC markers; GENE therapy; IMMUNOTHERAPY
- Publication
Gene Therapy, 2001, Vol 8, Issue 18, p1427
- ISSN
0969-7128
- Publication type
Article
- DOI
10.1038/sj.gt.3301530