In Vivo Selection of CD4+ T Cells Transduced with a Gamma-Retroviral Vector Expressing a Single-Chain Intrabody Targeting HIV-1 Tat
In Vivo Selection of CD4+ T Cells Transduced with a Gamma-Retroviral Vector Expressing a Single-Chain Intrabody Targeting HIV-1 Tat
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DOI：
10.1089/hum.2011.184
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发表时间：
2012-09-01
影响因子：
4.2
通讯作者：
Marasco, Wayne A.
中科院分区：
文献类型：
--
作者：
Braun, Stephen E.;Taube, Ran;Marasco, Wayne A.
我们评估了一种抗人类免疫缺陷病毒（HIV）的体内抗体（细胞内抗体）在恒河猴嵌合猴免疫缺陷病毒(SIV)/HIV （SHIV）感染后促进CD4（+）细胞存活的潜力。在优化刺激和转导条件后，纯化的CD4(+) T细胞用表达抗hiv -1 Tat或对照单链体内的galv假型逆转录病毒载体转导。体外体内基因标记效率很高，平均每个CD4（+）细胞有4个拷贝。将工程化的自体CD4（+）细胞再输注到两只猕猴体内后，检测到高水平的基因标记（峰值分别为外周血单核细胞（PBMCs）的0.6%和6.8%以及淋巴结细胞的0.3%和2.2%）。细胞输注一周后，用SHIV 89.6p刺激动物，并评估体内抗hiv Tat促进细胞存活的能力。在两种动物中，基因修饰的CD4(+) T细胞的频率逐渐下降，同时CD4（+）细胞的丢失和病毒载量的升高。然而，在8周和21周的时间内，表达治疗性抗tat抗体的CD4(+) T细胞比表达对照体内抗tat抗体的CD4(+) T细胞表现出相对的生存优势。在一种动物中，抗tat转导细胞的这种生存益处与病毒载量的减少有关。总之，这些结果表明，逆转录病毒介导的体内抗tat在pbmc和外周组织中提供了显著水平的基因标记，并提高了转导细胞在体内的相对存活率。
We evaluated the potential of an anti-human immunodeficiency virus (HIV) Tat intrabody (intracellular antibody) to promote the survival of CD4(+) cells after chimeric simian immunodeficiency virus (SIV)/HIV (SHIV) infection in rhesus macaques. Following optimization of stimulation and transduction conditions, purified CD4(+) T cells were transduced with GaLV-pseudotyped retroviral vectors expressing either an anti-HIV-1 Tat or a control single-chain intrabody. Ex vivo intrabody-gene marking was highly efficient, averaging four copies per CD4(+) cell. Upon reinfusion of engineered autologous CD4(+) cells into two macaques, high levels of gene marking (peak of 0.6% and 6.8% of peripheral blood mononuclear cells (PBMCs) and 0.3% or 2.2% of the lymph node cells) were detected in vivo. One week post cell infusion, animals were challenged with SHIV 89.6p and the ability of the anti-HIV Tat intrabody to promote cell survival was evaluated. The frequency of genetically modified CD4(+) T cells progressively decreased, concurrent with loss of CD4(+) cells and elevated viral loads in both animals. However, CD4(+) T cells expressing the therapeutic anti-Tat intrabody exhibited a relative survival advantage over an 8- and 21-week period compared with CD4(+) cells expressing a control intrabody. In one animal, this survival benefit of anti-Tat transduced cells was associated with a reduction in viral load. Overall, these results indicate that a retrovirus-mediated anti-Tat intrabody provided significant levels of gene marking in PBMCs and peripheral tissues and increased relative survival of transduced cells in vivo.