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Titolo:
FLT3 ligand preserves the uncommitted CD34(+) CD38(-) progenitor cells during cytokine prestimulation for retroviral transduction
Autore:
Nielsen, SD; Husemoen, LLN; Sorensen, TU; Gram, GJ; Hansen, JES;
Indirizzi:
Univ Copenhagen, Hvidovre Hosp, Infect Dis Lab, DK-2650 Hvidovre, Denmark Univ Copenhagen Hvidovre Denmark DK-2650 Lab, DK-2650 Hvidovre, Denmark
Titolo Testata:
JOURNAL OF HEMATOTHERAPY & STEM CELL RESEARCH
fascicolo: 5, volume: 9, anno: 2000,
pagine: 695 - 701
SICI:
1525-8165(200010)9:5<695:FLPTUC>2.0.ZU;2-W
Fonte:
ISI
Lingua:
ENG
Soggetto:
HEMATOPOIETIC STEM-CELLS; IMMUNODEFICIENCY-VIRUS TYPE-1; PERIPHERAL-BLOOD LYMPHOCYTES; EFFICIENCY GENE-TRANSFER; MURINE LEUKEMIA-VIRUS; RIBOSOME ENTRY SITE; IN-VIVO; ANTIRETROVIRAL THERAPY; MEDIATED TRANSDUCTION; INFECTED PATIENTS;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Clinical Medicine
Life Sciences
Citazioni:
38
Recensione:
Indirizzi per estratti:
Indirizzo: Nielsen, SD Univ Copenhagen, Hvidovre Hosp, Infect Dis Lab, 144 KettegaardAlle 30, DK-2650 Hvidovre, Denmark Univ Copenhagen 144 Kettegaard Alle 30 Hvidovre Denmark DK-2650
Citazione:
S.D. Nielsen et al., "FLT3 ligand preserves the uncommitted CD34(+) CD38(-) progenitor cells during cytokine prestimulation for retroviral transduction", J HEMATH ST, 9(5), 2000, pp. 695-701

Abstract

Before stem cell gene therapy can be considered for clinical applications,problems regarding cytokine prestimulation remain to be solved. In this study, a retroviral vector carrying the genes for the enhanced version of green fluorescent protein (EGFP) and neomycin resistance (neo(r)) was used fortransduction of CD34(+) cells. The effect of cytokine prestimulation on transduction efficiency and the population of uncommitted CD34(+)CD38(-) cells was determined. CD34(+) cells harvested from umbilical cord blood were kept in suspension cultures and stimulated with combinations of the cytokinesstem cell factor (SCF), FLT3 ligand, interleukin-3 (IL-3), IL-6, and IL-7 prior to transduction. Expression of the two genes was assessed by flow cytometry and determination of neomycin-resistant colonies in a selective colony-forming unit (CFU) assay, respectively. The neomycin resistance gene wasexpressed in a higher percentage of cells than the EGFP gene, but there seemed to be a positive correlation between expression of the two genes. The effect of cytokine prestimulation was therefore monitored using EGFP as marker for transduction. When SCF was compared to SCF in combination with morepotent cytokines, highest transduction efficiency was found with SCF and IL-3 and IL-6 (5.05% +/- 0.80 versus 2.66% +/- 0.53 with SCF alone, p = 0.04). However, prestimulation with SCF in combination with IL-3 and IL-6 also reduced the percentage of CD34(+) cells (p = 0.02). Then, prestimulation with SCF and FLT3 ligand was compared. Significant difference in transductionefficiency was not found. Interestingly, FLT3 ligand seemed to preserve the population of CD34(+)CD38(-) cells compared to SCF (16.56% +/- 2.02 versus 9.39% +/- 2.35, p = 0.03). In conclusion, prestimulation with potent cytokine combinations increased the transduction efficiency, but reduced the fraction of CD34(+) cells. Importantly, the use of FLT3 ligand seemed to preserve the population of uncommitted cells.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 24/11/20 alle ore 14:37:16