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Titolo:
Retrovirus-mediated gene transfer into T cells: 95% transduction efficiency without further in vitro selection
Autore:
Movassagh, M; Boyer, O; Burland, MC; Leclercq, V; Klatzmann, D; Lemoine, FM;
Indirizzi:
CHU Pitie Salpetriere, CERVI, CNRS, ESA 7087, F-75651 Paris, France CHU Pitie Salpetriere Paris France F-75651 A 7087, F-75651 Paris, France
Titolo Testata:
HUMAN GENE THERAPY
fascicolo: 8, volume: 11, anno: 2000,
pagine: 1189 - 1200
SICI:
1043-0342(20000520)11:8<1189:RGTITC>2.0.ZU;2-H
Fonte:
ISI
Lingua:
ENG
Soggetto:
PERIPHERAL-BLOOD LYMPHOCYTES; FIBRONECTIN FRAGMENTS; TARGET-CELLS; CORD-BLOOD; HIGH-LEVEL; LEUKEMIA; REPERTOIRE; THERAPY; CD4(+); AGE;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
23
Recensione:
Indirizzi per estratti:
Indirizzo: Lemoine, FM CHU Pitie Salpetriere, CERVI, CNRS, ESA 7087, 83 Blvd Hop, F-75651 Paris, France CHU Pitie Salpetriere 83 Blvd Hop Paris France F-75651 France
Citazione:
M. Movassagh et al., "Retrovirus-mediated gene transfer into T cells: 95% transduction efficiency without further in vitro selection", HUM GENE TH, 11(8), 2000, pp. 1189-1200

Abstract

This study was designed to retrovirally transduce T cells by a protocol that would be simple, short, cost effective, applicable for clinical use, andefficient enough to avoid further selection of transduced T cells. Becauseretrovirally mediated infection is depending on the cell cycle, we first optimized the conditions for activating T cells in the presence of immobilized CD3 monoclonal antibodies and recombinant interleukin 2. Cell cycle analysis indicated that CD8(+) and total T cells reach a maximum of cycling within 4 days whereas CD4(+) T cells attain their maximum of cycling only by day 6. Taking into account these data, CD4(+), CD8(+) and total T cells merepreactivated for 5 and 3 days, respectively, and then infected for 24 hr with supernatant containing retrovirus pseudotyped with gibbon-ape leukemia virus envelope, using a cell centrifugation protocol. Results show that approximately 95% of CD4(+), CD8(+), and total T cells can be transduced, thistransduction efficiency being significantly higher than that obtained,vithamphotropic retrovirus vectors. Furthermore, under permanent growth stimulation, transduced T cells can be expanded similar to 1000-fold in 4 weeks of culture with maintenance of transgene expression. Ho cr ever, Immunoscopeanalysis revealed alterations of T cell repertoire diversity after 2-3 weeks in culture that was not due to retroviral transduction per se. Overall, these data provide evidence that T cells can be transduced at levels that may alleviate the need for both further selection of transduced cells and invitro expansion, thereby preserving the repertoire diversity of the transduced T cells to be reinfused.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 29/09/20 alle ore 07:39:19