Efficient infection of a human T-cell line and of human primary peripheral blood leukocytes with a pseudotyped retrovirus vector.
Efficient infection of a human T-cell line and of human primary peripheral blood leukocytes with a pseudotyped retrovirus vector.
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用假型逆转录病毒载体有效感染人类 T 细胞系和人类原代外周血白细胞。
DOI：
10.1073/pnas.93.21.11842
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发表时间：
1996
影响因子：
11.1
通讯作者：
Friedmann,T
中科院分区：
文献类型：
--
作者：
Sharma,S;Cantwell,M;Kipps,TJ;Friedmann,T
外周血淋巴细胞（PBL）是人类基因治疗中基因转移研究的重要靶点。然而，目前还没有可重复的有效方法将外源的、潜在的治疗基因转移到这些细胞中。虽然来源于鼠逆转录病毒的载体是最广泛使用的系统，但它们在淋巴细胞中的感染效率低，需要在感染后延长体外培养和选择以获得有用数量的遗传修饰细胞。我们先前报道了用水泡性口炎G糖蛋白（VSV-G）包膜假型化的逆转录病毒载体可以感染多种细胞类型，并且可以浓缩至大于10（9）感染单位/ml的滴度。在本研究中，我们研究了表达鼠细胞表面蛋白B7-1的嗜中性和假型载体感染人T细胞系Jurkat或人血淋巴细胞的能力。转导的Jurkat细胞的有限稀释分析表明，假型化载体在感染T细胞方面比在相同感染复数（moi）下使用的亲嗜性载体显著更有效。为了鉴定对PBL的转导效率，我们检测了感染后48至72小时B7-1表面标志物的细胞表面表达水平。用假型化载体的PBL的转导效率随着moi的增加而线性增加，在moi为40时达到约16-32%的最大值。用VSV-G假型病毒感染T细胞系和血淋巴细胞的这种相对高的效率表明，这种经修饰的假型逆转录病毒载体可以是用于研究各种遗传或肿瘤疾病的基因治疗的有用试剂。
Peripheral blood lymphocytes (PBLs) are an important target for gene transfer studies aimed at human gene therapy. However, no reproducibly efficient methods are currently available to transfer foreign, potentially therapeutic genes into these cells. While vectors derived from murine retroviruses have been the most widely used system, their low infection efficiency in lymphocytes has required prolonged in vitro culturing and selection after infection to obtain useful numbers of genetically modified cells. We previously reported that retroviral vectors pseudotyped with vesicular stomatitis G glycoprotein (VSV-G) envelope can infect a wide variety of cell types and can be concentrated to titers of greater than 10(9) infectious units/ml. In this present study, we examined the ability of amphotropic and pseudotyped vectors expressing a murine cell surface protein, B7-1, to infect the human T-cell line Jurkat or human blood lymphocytes. Limiting dilution analysis of transduced Jurkat cells demonstrated that the pseudotyped vector is significantly more efficient in infecting T cells than an amphotropic vector used at the same multiplicity of infection (moi). To identify the transduction efficiency on PBLs, we examined the levels of cell surface expression of the B7-1 surface marker 48 to 72 hr after infection. The transduction efficiency of PBLs with the pseudotyped vector increased linearly with increasing moi to a maximum of approximately 16-32% at an moi of 40. This relatively high efficiency of infection of a T-cell line and of blood lymphocytes with VSV-G pseudotyped virus demonstrates that such modified pseudotyped retrovirus vectors may be useful reagents for studies of gene therapy for a variety of genetic or neoplastic disorders.
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DOI：
10.1073/pnas.91.20.9564
发表时间：
1994-09-27
影响因子：
11.1
作者：
YEE, JK;MIYANOHARA, A;FRIEDMANN, T
通讯作者：
FRIEDMANN, T
影响因子：
56.9
作者：
T. Friedmann
通讯作者：
T. Friedmann
外周血淋巴细胞作为逆转录病毒载体介导的基因转移的靶细胞
影响因子：
20.3
作者：
F. Mavilio;G. Ferrari;S. Rossini;N. Nobili;C. Bonini;G. Casorati;C. Traversari;C. Bordignon
通讯作者：
C. Bordignon
DOI：
10.1073/pnas.90.17.8033
发表时间：
1993-09-01
影响因子：
11.1
作者：
BURNS, JC;FRIEDMANN, T;YEE, JK
通讯作者：
YEE, JK
产生具有非常广泛宿主范围的高滴度假型逆转录病毒载体。
影响因子：
--
作者：
Yee,JK;Friedmann,T;Burns,JC
通讯作者：
Burns,JC