HIV-1 vector mediated gene therapy for HIV-1 infection

描述(由申请人提供)：本研究的长期目标是 确定基于HIV-I的载体是否可以防止艾滋病的进展 感染艾滋病毒的人。我们已经证明，携带抗艾滋病毒的艾滋病毒载体 反义基因有效载荷(VRX496)可通过以下途径转导原代人CD4T细胞 两周后FACS检测转导效率超过90% 转导。以这种方式转导的CD4T细胞直接受到攻击 WT-HIV，并被证明在正常的初级CD4T细胞中抑制HIV-I复制 单元格超过p24的三个对数单位。类似的抑制作用 当转导细胞被不同种类的病毒攻击时，WT-HIV复制 艾滋病毒的主要毒株。值得注意的是，原代CD4T细胞通过 VRX496被发现对生产性HIV感染具有抵抗力，因此具有 与未用载体修饰的细胞相比，存活优势。此外， 初步数据显示，VRX496在CD4T细胞中具有类似的作用 来自一名感染艾滋病毒的捐赠者。临床翻译的一个重要目标 这类载体将表征它们对细胞的抑制作用 存在于来自HIV感染者的细胞中的致病病毒。这个 这个项目的具体目标是充分描述抑制效应的特征 VRX496在来自HIV感染者捐赠者的原代人类CD4T细胞中的表达 根据他们的临床状态而不同。从感染HIV的供者体内分离出CD4T细胞 病毒载量、CD4计数和HAART治疗的不协调将通过 并对VRX496的生物学和抗HIV作用进行了表征。表征 VRX496在多种HIV感染者细胞中的作用 患者的体外实验将为未来的合理设计提供重要数据 活体临床研究。 建议的商业应用： 基于HIV-1的载体可以非常高效地将有效载荷基因输送到原代人类细胞中。当抗HIV有效载荷基因被传递到原代CD4+T细胞，然后感染wt-HIV时，通过ELISA抗原捕获试验，病毒复制可以被超过2个对数单位的p24所抑制。这项技术的潜在商业应用之一是治疗感染艾滋病的个人。这项技术还可以用于癌症等其他疾病的基因治疗，因为这种载体可以有效地转导多种类型的细胞。

DESCRIPTION (Provided by the applicant): The long-term goal of this study is to determine whether HIV-I based vectors can prevent AIDS disease progression in HIV-infected individuals. We have shown that an HIV vector carrying an anti-HIV antisense genetic payload (VRX496) can transduce primary human CD4 T cells by over 90 percent transduction efficiency, as measured by FACS two weeks after transduction. CD4 T cells transduced in this manner were directly challenged with wt-HIV and shown to inhibit HIV-I replication in normal primary CD4 T cells by over three logarithmic units of p24. Similar inhibitory effects on wt-HIV replication were seen when transduced cells were challenged with various primary strains of HIV. Significantly, primary CD4 T cells transduced with VRX496 were found to be resistant to productive HIV infection and hence, had a survival advantage over cells not modified with the vector. Furthermore, preliminary data have shown similar effects of VRX496 in CD4 T cells derived from one HIV-infected donor. An important goal for the clinical translation of this class of vectors would be to characterize their inhibitory effects on the pathogenic virus present in cells derived from HIV-infected individuals. The specific goal of this project is to fully characterize the inhibitory effects of VRX496 in primary human CD4 T cells derived from HIV-infected donors that differ by their clinical status. CD4 T cells isolated from HIV infected donors discordant in viral load, CD4 count and HAART therapy will be transduced with VRX496 and the biological and anti-HIV effects characterized. Characterization of the effects of VRX496 in the cells from a wide variety of HIV-infected patients ex vivo will provide important data for rational design of future in vivo clinical studies. PROPOSED COMERCIAL APPLICATIONS: HIV-1 based vectors can deliver payload genes into primary human cells with very high efficiency. When a anti-HIV payload gene is deliv ered into primary CD4+ T cells that are then infected with wt-HIV, viral replication can be inhibited by over 2 logarithmic units of p24, as measured by ELISA antigen capture assay. One of the potential commercial applications of the technology is for the treatment of individuals infected with AIDS. The technology can also be used for gene therapy of other diseases such as cancer because this class of vectors can efficiently transduce many cell types.