Catalogo Articoli (Spogli Riviste)

OPAC HELP

Titolo:
Systematic analysis of clinically applicable conditions leading to a high efficiency of transduction and transgene expression in human T cells
Autore:
Lamana, ML; Segovia, JC; Guenechea, G; Bueren, JA;
Indirizzi:
CIEMAT Fdn, Gene Therapy Programme, Madrid 28040, Spain CIEMAT Fdn Madrid Spain 28040 ene Therapy Programme, Madrid 28040, Spain
Titolo Testata:
JOURNAL OF GENE MEDICINE
fascicolo: 1, volume: 3, anno: 2001,
pagine: 32 - 41
SICI:
1099-498X(200101/02)3:1<32:SAOCAC>2.0.ZU;2-J
Fonte:
ISI
Lingua:
ENG
Soggetto:
PERIPHERAL-BLOOD LYMPHOCYTES; MEDIATED GENE-TRANSFER; TUMOR-INFILTRATING LYMPHOCYTES; SCAFFOLD ATTACHMENT REGION; GRAFT-VERSUS-LEUKEMIA; THYMIDINE KINASE GENE; RETROVIRAL VECTOR; BONE-MARROW; FIBRONECTIN FRAGMENTS; HEMATOPOIETIC-CELLS;
Keywords:
human T lymphocytes; retroviral vectors; gene transfer; nerve growth factor receptor; transgene expression;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
40
Recensione:
Indirizzi per estratti:
Indirizzo: Bueren, JA CIEMAT Fdn, Gene Therapy Programme, Ave Complutense 22, Madrid 28040, Spain CIEMAT Fdn Ave Complutense 22 Madrid Spain 28040 28040, Spain
Citazione:
M.L. Lamana et al., "Systematic analysis of clinically applicable conditions leading to a high efficiency of transduction and transgene expression in human T cells", J GENE MED, 3(1), 2001, pp. 32-41

Abstract

Background The transduction of human peripheral blood T cells with retroviral vectors constitutes an attractive approach for the correction of a number of genetic diseases. In this study we have conducted a systematic analysis of the relevance of a large number of parameters currently considered toaffect the transduction of, and transgene expression in, human T cells. Methods Retroviral vectors encoding the human nerve growth factor receptor(NGFR) were used for transducing human T cells from normal volunteers. Theproportion of T cells that expressed the marker transgene was determined by flow cytometry using anti-NGFR antibodies. Results Spinoculation and static fibronectin (FN)-assisted infections improved to a similar extent the transduction efficiency of PHA/IL-2 stimulatedT cells, when compared with samples subjected to standard static infections. When immobilized anti-CD3 (anti-CD3i) or anti-CD3i/28i-stimulated T cells were considered, static infections in FN-coated plates were reproducibly more efficient than spinoculation infections performed on FN-uncoated plates. Under optimized manipulation conditions (three infection cycles of anti-CD3i/28i-stimulated T cells in FN-coated plates) the total number of NGFR(+) T cells harvested after 7 days of incubation represented, on average, twice the total number of T cells seeded at Day 0, and up to 95% of the human T cells efficiently expressed the marker transgene. Similar results were obtained regardless of whether samples were manipulated in medium supplemented with fetal bovine serum or with heat-inactivated autologous serum. Conclusions Our study offers new experimental conditions for the transduction of human T cells, with obvious implications for the development of genetherapy protocols. Copyright (C) 2000 John Wiley & Sons, Ltd.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 22/09/20 alle ore 20:11:27