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Titolo:
Lentiviral and murine retroviral transduction of T cells for expression ofhuman CD40 ligand
Autore:
Barry, SC; Seppen, J; Ramesh, N; Foster, JL; Seyama, K; Ochs, HD; Garcia, JV; Osborne, WRA;
Indirizzi:
Univ Washington, Dept Pediat, Seattle, WA 98195 USA Univ Washington Seattle WA USA 98195 , Dept Pediat, Seattle, WA 98195 USA Univ Texas, SW Med Ctr, Dept Internal Med, Dallas, TX 75235 USA Univ Texas Dallas TX USA 75235 r, Dept Internal Med, Dallas, TX 75235 USA
Titolo Testata:
HUMAN GENE THERAPY
fascicolo: 2, volume: 11, anno: 2000,
pagine: 323 - 332
SICI:
1043-0342(20000120)11:2<323:LAMRTO>2.0.ZU;2-D
Fonte:
ISI
Lingua:
ENG
Soggetto:
HYPER-IGM SYNDROME; HUMAN ADENOSINE-DEAMINASE; LONG-TERM EXPRESSION; HUMAN GENE-THERAPY; FIBRONECTIN FRAGMENTS; SURFACE RECEPTORS; MUTATIONS; VECTORS; VIRUS; GP39;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
36
Recensione:
Indirizzi per estratti:
Indirizzo: Osborne, WRA Univ Washington, Dept Pediat, 1959 NE Pacific St,MS 356320, Seattle, WA 98195 USA Univ Washington 1959 NE Pacific St,MS 356320 Seattle WA USA 98195
Citazione:
S.C. Barry et al., "Lentiviral and murine retroviral transduction of T cells for expression ofhuman CD40 ligand", HUM GENE TH, 11(2), 2000, pp. 323-332

Abstract

Efficient transduction of primary human T cells is an important goal toward treating a number of genetic defects. Patient T cells could be harvested by leukapheresis, transduced, and returned to the donor. A wide range of secreted or cell surface therapeutic proteins may be delivered in this way. The ability to produce antibodies is the consequence of interactions betweenT cells and B cells and lack of expression of CD40 ligand (CD40L) on T cells causes X-linked hyper-IgM syndrome (XHIM). We are investigating deliveryof a normal CD40 ligand to treat this disorder, We tested promoters driving the expression of either reporter genes such as enhanced green fluorescent protein (eGFP) or human CDC40L. Using murine retroviruses, the best able to drive gene expression in T cells was the cytomegalovirus (CMV) promoter enhancer element; however, transduction efficiency was low. To achieve efficient, high-level gene expression we tested lentiviral gene delivery vectors. At a low multiplicity of infection (MOI) (0.5-2) a large fraction of target cells was transduced by lentiviral vectors (40-93%), and the strength of gene expression was high, as determined by flow cytometric analysis. We monitored the expression of eGFP or human CD40L on T cell lines and untransformed primary human T cells from normal and CD40L-deficient patients. We achieved efficient gene expression without an extended exposure to virus, andwithout the need for selection. These results are encouraging for efficient lentivirus-mediated transduction of refractory human cells to achieve therapeutic gene delivery.

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Documento generato il 29/09/20 alle ore 06:55:23