Sustained long-term engraftment and transgene expression of peripheral blood CD34+ cells transduced with third-generation lentiviral vectors.
As mobilized peripheral blood (MPB) represents an attractive cell source for gene therapy, we investigated the ability of third-generation lentiviral vectors (LVs) to transfer the enhanced green fluorescent protein gene into MPB CD34(+) cells in culture conditions allowing expansion of transplantable human hematopoietic stem cells. To date, few studies have reported transduction of MPB cells with vesicular stomatitis virus G pseudotyped LVs. The critical issue remains whether primitive, hematopoietic repopulating cells have, indeed, been transduced. In vitro (5 weeks' culture in FLT3 ligand + thrombopoietin + stem cell factor + interleukin 6) and in vivo (serial transplantation in NOD/SCID mice) experiments show that MPB CD34(+) cells can be effectively long-term transduced by LV and maintain their proliferation, self-renewal, and multilineage differentiation potentials. We show that expansion following transduction improves the engraftment of transduced MPB CD34(+) (4.6-fold expansion of SCID repopulating cells by limiting dilution studies). We propose ex vivo expansion after transduction as an effective tool to improve gene therapy protocols with MPB. Disclosure of potential conflicts of interest is found at the end of this article.
Tesio
Melania
M
Department of Oncological Sciences, Laboratory of Clinical Oncology, University of Torino Medical School, Institute for Cancer Research and Treatment, Candiolo, Torino, Italy.
Gammaitoni
Loretta
L
Gunetti
Monica
M
Leuci
Valeria
V
Pignochino
Ymera
Y
Jordaney
Noela
N
Capellero
Sonia
S
Cammarata
Cristina
C
Caione
Luisa
L
Migliaretti
Giuseppe
G
Fagioli
Franca
F
Tabilio
Antonio
A
Aglietta
Massimo
M
Piacibello
Wanda
W
D000818:Animals; D018952:Antigens, CD34 / Q000032:analysis*; D001773:Blood Cells / Q000502:physiology; D001854:Bone Marrow Cells / Q000502:physiology; D005434:Flow Cytometry; D017930:Genes, Reporter; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000494:pharmacology; D049452:Green Fluorescent Proteins / Q000235:genetics; D015497:HIV-1 / Q000235:genetics*; D019650:Hematopoietic Stem Cell Mobilization; D006801:Humans; D016130:Immunophenotyping; D007377:Interleukin-3 / Q000494:pharmacology; D051379:Mice; D016513:Mice, SCID; D011994:Recombinant Proteins / Q000494:pharmacology; D013926:Thrombopoietin / Q000494:pharmacology
D018952:Antigens, CD34; D007377:Interleukin-3; D011994:Recombinant Proteins; D049452:Green Fluorescent Proteins; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D013926:Thrombopoietin