Therapeutic Dendritic Cell Vaccine for HIV

项目摘要 HIV感染者体内存在长期的潜伏感染的CD4T细胞需要 他们终生都在接受抗逆转录病毒药物治疗。少数精英控制员将病毒载量维持在 在没有抗逆转录病毒治疗的情况下，通过标准临床检测进行检测的可能性很大。这样的人有能力 抑制病毒复制与抗病毒溶解T细胞有关，抗病毒T细胞作为一种 持续的受体刺激和检查点激活的结果。该项目将开发一种治疗方法 慢病毒载体树突状细胞疫苗增强CD8 T细胞应答和逆转 精疲力竭，以实现功能治愈，在此过程中，病毒复制被抑制，而不需要抗病毒治疗。这个 疫苗是基于慢病毒载体在病毒粒子中传递的，该病毒粒子包含SIV辅助蛋白VPX用于高 高效DC转导和表达HIV-1抗原、免疫刺激细胞因子和检查点 抑制剂。抗原将被表达为一种融合蛋白，允许在TAP上不依赖于TAP呈现 I类主要组织相容性蛋白。载体诱导抗病毒T细胞反应的能力将是 在体外和体内进行测试。艾滋病患者外周血单个核细胞来源的DC将被转化 并对其激活和扩增自身HIV特异性T细胞的能力进行了测试 测定CD8T细胞表位特异性和反应频率的ELISPOT法 整个HIV基因组。载体转导的树突状细胞诱导抗原特异性T细胞和 抑制病毒载量将在人源化的小鼠模型中确定。疫苗逆转T细胞的能力 将评估淋巴细胞性脉络膜脑膜炎的细胞耗竭和刺激保护性反应。 病毒小鼠模型。此外，含有VPX的慢病毒载体将在工程免疫中进行测试 树突状细胞体内长期表达抗病毒蛋白的方法。

Project Summary The presence of a long-lived reservoir of latently infected CD4 T cells in HIV infected individuals requires that they remain on antiretroviral drug regimens life-long. Rare elite controllers maintain virus loads below the level of detection by standard clinical assays without antiretroviral treatment. The ability of such individuals to suppress virus replication is associated with antiviral cytolytic T cells that are resistant to exhaustion as a result of continuous receptor stimulation and checkpoint activation. The project will develop a therapeutic lentiviral vector-based dendritic cell (DC) vaccine that enhances CD8 T cell responses and reverses exhaustion to achieve a functional cure in which virus replication is suppressed without antiviral therapy. The vaccine is based on lentiviral vectors delivered in virions that contain the SIV accessory protein Vpx for high efficiency DC transduction and that express HIV-1 antigen, an immunostimulatory cytokine and a checkpoint inhibitor. The antigen will be expressed as a fusion protein that allows for TAP-independent presentation on class I major histocompatibility proteins. The ability of the vectors to induce anti-viral T cell responses will be tested in vitro and in vivo. AIDS patient peripheral blood mononuclear cell-derived DCs will be transduced with the vectors and tested for their ability to activate and expand autologous HIV-specific T cells in a high throughput ELISPOT assay that determines the epitope specificity and frequency of responding CD8 T cells across the HIV genome. The ability of vector-transduced DCs to induce antigen specific T cells and to suppress virus load will be determined in a humanized mouse model. The ability of the vaccine to reverse T cell exhaustion and to stimulate protective responses will be evaluated in the lymphocytic choriomeningitis virus mouse model. In addition, Vpx-containing lentiviral vectors will be tested in an engineered immunity approach for the long-term in vivo expression of antiviral proteins in DCs.