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Titolo:
High-efficiency gene transfer into rhesus macaque primary T lymphocytes bycombining 32 degrees C centrifugation and CH-296-coated plates: Effect of gene transfer protocol on T cell homing receptor expression
Autore:
Zhou, P; Lee, J; Moore, P; Brasky, KM;
Indirizzi:
SW Fdn Biomed Res, Dept Virol & Immunol, San Antonio, TX 78245 USA SW Fdn Biomed Res San Antonio TX USA 78245 nol, San Antonio, TX 78245 USA SW Fdn Biomed Res, Dept Lab Anim Med, San Antonio, TX 78245 USA SW Fdn Biomed Res San Antonio TX USA 78245 Med, San Antonio, TX 78245 USA
Titolo Testata:
HUMAN GENE THERAPY
fascicolo: 15, volume: 12, anno: 2001,
pagine: 1843 - 1855
SICI:
1043-0342(20011015)12:15<1843:HGTIRM>2.0.ZU;2-I
Fonte:
ISI
Lingua:
ENG
Soggetto:
HUMAN-IMMUNODEFICIENCY-VIRUS; APE LEUKEMIA-VIRUS; PERIPHERAL-BLOOD LYMPHOCYTES; HEMATOPOIETIC STEM-CELLS; L-SELECTIN; FIBRONECTIN FRAGMENTS; RETROVIRAL VECTORS; INFECTED PATIENTS; MESSENGER-RNA; CROSS-LINKING;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
55
Recensione:
Indirizzi per estratti:
Indirizzo: Zhou, P SW Fdn Biomed Res, Dept Virol & Immunol, 7620 NW Loop 410, San Antonio, TX78227 USA SW Fdn Biomed Res 7620 NW Loop 410 San Antonio TX USA 78227 27 USA
Citazione:
P. Zhou et al., "High-efficiency gene transfer into rhesus macaque primary T lymphocytes bycombining 32 degrees C centrifugation and CH-296-coated plates: Effect of gene transfer protocol on T cell homing receptor expression", HUM GENE TH, 12(15), 2001, pp. 1843-1855

Abstract

Although steady progress has been made in transducing human T lymphocytes by Moloney murine leukemia virus (Mo-MuLV)-based vectors, few studies have been done to define ex vivo gene transfer protocols to transduce rhesus macaque primary T lymphocytes. Given the fact that simian immunodeficiency virus (SIV) infection in rhesus macaque is a well-characterized model for human immunodeficiency virus (HIV), it is of great interest to develop an efficient protocol to transduce rhesus macaque primary T cells. In this study, we have used MuLV-10A1-pseudotyped retrovirus expressing enhanced green fluorescent protein (EGFP) to evaluate a number of ex vivo gene transfer protocols in rhesus macaque primary T lymphocytes. Our objectives in designing these protocols were (1) to test whether higher efficiency gene transfer could be obtained by combining two previously defined protocols, centrifugationat 32 degreesC and the CH-296-coated plate; and (2) to study the effect ofan ex vivo gene transfer protocol on the expression of lymphocyte homing receptors L-selectin and alpha4 beta7 and alpha4 beta1 integrins. From sevenindependent experiments we demonstrate by flow cytometry analyses of EGFP expression that whereas centrifugation at 32 degreesC or the fibronectin fragment CH-296-coated plate protocol alone yielded 10-14% transduction efficiency, combining these two protocols resulted in 28.1-51.2% transduction efficiency. EGFP in transduced cells was expressed highly throughout the 14 days of posttransduction expansion. Our results also demonstrate that whereas the transduction procedure per se did not significantly alter the expression of lymphocyte homing receptors, anti-CD3 and anti-CD28 antibody stimulation profoundly reduced the expression of L-selectin. The selective reduction of L-selectin may result in significant in vivo consequences if transduced cells are infused.

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Documento generato il 24/09/20 alle ore 21:28:49