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Titolo:
Establishment of an optimised gene transfer protocol for human primary T lymphocytes according to clinical requirements
Autore:
Ayuk, FA; Li, Z; Kuhlcke, K; Lindemann, C; Schade, UM; Eckert, HG; Zander, AR; Fehse, B;
Indirizzi:
Univ Hamburg, Hosp Eppendorf, D-20251 Hamburg, Germany Univ Hamburg Hamburg Germany D-20251 Eppendorf, D-20251 Hamburg, Germany EUFETS GmbH, Idar Oberstein, Germany EUFETS GmbH Idar Oberstein GermanyEUFETS GmbH, Idar Oberstein, Germany
Titolo Testata:
GENE THERAPY
fascicolo: 10, volume: 6, anno: 1999,
pagine: 1788 - 1792
SICI:
0969-7128(199910)6:10<1788:EOAOGT>2.0.ZU;2-T
Fonte:
ISI
Lingua:
ENG
Soggetto:
APE LEUKEMIA-VIRUS; FIBRONECTIN FRAGMENTS; RETROVIRAL VECTORS; CELLS; EFFICIENCY; INFECTION; THERAPY; TRANSDUCTION;
Keywords:
gene therapy; retroviral vectors; T lymphocytes;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
19
Recensione:
Indirizzi per estratti:
Indirizzo: Fehse, B Univ Hamburg, Hosp Eppendorf, Martinistr 52, D-20251 Hamburg, Germany Univ Hamburg Martinistr 52 Hamburg Germany D-20251 burg, Germany
Citazione:
F.A. Ayuk et al., "Establishment of an optimised gene transfer protocol for human primary T lymphocytes according to clinical requirements", GENE THER, 6(10), 1999, pp. 1788-1792

Abstract

Current gene therapeutic protocols directed towards the treatment of inherited disorders (eg ADA-SCID) and viral infections (eg AIDS), as well as adoptive immunotherapy approaches are based on the use of genetically modifiedlymphocytes. Since only insufficient transduction of T cells is obtained using existing techniques, the development of more efficient gene transfer protocols into these cells is of great importance. We present here a protocol for the highly efficient transduction of human primary T cells at high densities (1 x 10(6)/ml) by retroviral infection. Using retroviral vectors encoding a truncated human low-affinity nerve growth factor receptor (Delta LNGFR) as a gene transfer marker, we obtained transduction frequencies of more than 70% of CD3(+) cells after two cycles of infection. Our protocol is based on the use of FBS-free media for both the production of retrovirus-containing supernatant and the cultivation of the primary T cells. Since the protocol presented here works just as efficiently under large-scale conditions, it may be easily adapted to clinical needs and 'good manufacturing practice' (GMP) standards.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 23/09/20 alle ore 15:29:36