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Titolo:
Pre-clinical evaluation of an in vitro selection protocol for the enrichment of transduced CD34(+) cell-derived human dendritic cells
Autore:
Evans, JT; Cravens, P; Gatlin, J; Kelly, PF; Lipsky, PE; Garcia, JV;
Indirizzi:
Univ Texas, SW Med Ctr, Dept Internal Med, Div Infect Dis Y9 206, Dallas, TX 75390 USA Univ Texas Dallas TX USA 75390 iv Infect Dis Y9 206, Dallas, TX 75390 USA Univ Texas, SW Med Ctr, Harold C Simmons Arthrit Res Ctr, Dept Internal Med, Dallas, TX 75390 USA Univ Texas Dallas TX USA 75390 r, Dept Internal Med, Dallas, TX 75390 USA St Jude Childrens Hosp, Dept Hematol, Memphis, TN 38105 USA St Jude Childrens Hosp Memphis TN USA 38105 ematol, Memphis, TN 38105 USA NIAMSD, Autoimmunol Branch, Bethesda, MD 20892 USA NIAMSD Bethesda MD USA20892 , Autoimmunol Branch, Bethesda, MD 20892 USA
Titolo Testata:
GENE THERAPY
fascicolo: 18, volume: 8, anno: 2001,
pagine: 1427 - 1435
SICI:
0969-7128(200109)8:18<1427:PEOAIV>2.0.ZU;2-P
Fonte:
ISI
Lingua:
ENG
Soggetto:
MEDIATED GENE-TRANSFER; CYTOTOXIC T-LYMPHOCYTES; RESISTANT BONE-MARROW; CORD-BLOOD; IN-VIVO; STEM-CELLS; DIHYDROFOLATE-REDUCTASE; NUCLEOSIDE TRANSPORT; HEMATOPOIETIC STEM; ANTITUMOR IMMUNITY;
Keywords:
dendritic cells; DHFR selection; gene therapy; lentivirus vectors; MLV vectors; immunotherapy;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
43
Recensione:
Indirizzi per estratti:
Indirizzo: Garcia, JV Univ Texas, SW Med Ctr, Dept Internal Med, Div Infect Dis Y9 206, Dallas, TX 75390 USA Univ Texas Dallas TX USA 75390 is Y9 206, Dallas, TX 75390 USA
Citazione:
J.T. Evans et al., "Pre-clinical evaluation of an in vitro selection protocol for the enrichment of transduced CD34(+) cell-derived human dendritic cells", GENE THER, 8(18), 2001, pp. 1427-1435

Abstract

The efficient genetic modification of CD34(+) 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(+) 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(+) cells and subsequentdifferentiation into mature DC. Each vector expressed both EGFP and the dominant selectable marker DHFRL22Y 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(+) cells. However, in vitro expansion and differentiation in the presence of GM-CSF, TNF-alpha, Flt-3L, SCF and IL-4 resulted in areduction in the percentage of DC expressing the transgene. Selection withTMTX 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.

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Documento generato il 02/12/20 alle ore 05:19:06