Inhibition of simian/human immunodeficiency virus replication in CD4+ T cells derived from lentiviral-transduced CD34+ hematopoietic cells
Inhibition of simian/human immunodeficiency virus replication in CD4+ T cells derived from lentiviral-transduced CD34+ hematopoietic cells
复制标题
DOI：
10.1016/j.ymthe.2005.07.698
复制
发表时间：
2005-12-01
影响因子：
12.4
通讯作者：
Johnson, RP
中科院分区：
文献类型：
--
作者：
Braun, SE;Wong, FE;Johnson, RP
我们研究了基于HIV-1的载体（VRX 494）编码937 bp反义HIV-1包膜序列抑制编码HIV-1包膜的嵌合SIV/HIV-1病毒复制的能力。VRX 494转导的CEMx 174细胞的攻击导致对HIV-1和几种SHIV毒株的有效抑制。为了评价VRX 494载体用于干细胞基因治疗的潜在功效，用VRX 494转导恒河猴CD 34（+）骨髓细胞，然后在胸腺基质上培养以诱导T细胞分化。对CD 34（+）细胞的转导条件进行了优化，以产生高转导效率和最小的有效感染复数。与对照相比，来自VRX 494转导的CD 34（+）细胞的纯化的CD 4（+）GFP（+）T细胞强烈抑制SHIV HXBC 2 P 3.2和SHIV 89.6P复制。VRX 494转导的T细胞克隆的Southern印迹分析揭示了每个细胞具有多个前病毒拷贝的细胞亚群。达特可上调VRX 494转导细胞中GFP和反义抑制剂的表达。VRX 494转导细胞中HIV-1包膜序列的分析揭示了与双链RNA依赖性腺苷脱氨酶介导的修饰一致的修饰。这些结果表明，猕猴/SHIV模型应作为一个有用的临床前模型，以评估这种慢病毒载体表达的HIV-1反义抑制剂的干细胞基因治疗艾滋病。
We examined the ability of a HIV-1-based vector (VRX494) encoding a 937-bp antisense HIV-1 envelope sequence to inhibit the replication of chimeric SIV/HIV-1 viruses encoding the HIV-1 envelope. Challenge of VRX494-transduced CEMx174 cells resulted in potent inhibition of HIV-1 and several SHIV strains. To evaluate the potential efficacy of the VRX494 vector for stem cell gene therapy, rhesus CD34(+) bone marrow cells were transduced with VRX494 and then cultured on thymus stroma to induce T cell differentiation. Transduction conditions for CD34(+) cells were optimized to yield high transduction efficiency with minimal effective multiplicity of infection. Purified CD4(+) GFP(+) T cells derived from VRX494-transduced CD34(+) cells strongly inhibited SHIV HXBC2P 3.2 and SHIV 89.6P replication compared to controls. Southern blot analysis of VRX494-transduced T cell clones revealed a subset of cells with multiple proviral copies per cell. Expression of GFP and the antisense inhibitor in VRX494-transduced cells was upregulated by Tat. Analysis of HIV-1 envelope sequences in VRX494-transduced cells revealed modifications consistent with those mediated by double-stranded RNA-dependent adenosine deaminase. These results indicate that the macaque/SHIV model should serve as a useful preclinical model to evaluate this lentiviral vector expressing an HIV-1 antisense inhibitor for stem cell gene therapy for AIDS.