Development of HIV vectors for antiviral and immune based therapies

这个临床前IPCP项目的主要目标是开发用于HIV疾病基因治疗的慢病毒载体。项目1的重点是开发针对造血细胞（包括干细胞和树突状细胞）的基于hiv的载体，用于抗病毒和免疫治疗。我们的具体目标是：1。为了建立最佳的HIV-2基因转移载体和稳定的包装线，将努力减少RCR的产生机会，同时优化载体滴度。2. 为了证明原代T细胞、巨噬细胞和CD34+细胞的后代通过慢病毒载体转导对HIV攻击的保护作用，这些研究将测试表达抗HIV-1和抗ccr5核酶的HIV-2载体。我们还将确定这些载体的转导是否会对这些靶细胞的细胞增殖、分化和/或功能产生任何有害影响。3. 为了研究慢病毒载体转导对树突状细胞功能和与T细胞相互作用的影响-树突状细胞（DC）既是HIV感染的促进者，也是防御者，因为它们可以向T细胞呈递病毒抗原和感染性病毒。最近的数据表明，虽然只有未成熟的DC可被HIV感染，但成熟的DC需要表达功能趋化因子受体CCR5和CXCR4才能分别将嗜m型和嗜t型病毒传播到CD4+ t细胞。我们将构建表达gag-pol抗原以及抗cc4 -5和/或抗cxcr4核酶的HIV-1载体，并确定这些载体的转导如何影响树突状细胞向CD4和CD8细胞呈递抗原或将HIV感染传播到CD4细胞的能力。除了这些目标之外，该项目还将与项目2和项目3密切合作，直接比较基于艾滋病毒和fiv的载体以及基于mlv的逆转录病毒载体在体外和体内的安全性和有效性。

The major goal of this preclinical IPCP project is the development of lentiviral vectors for gene therapy of HIV diseases. Project 1 focuses on developing HIV-based vectors that target hematopoietic cells, including stem cells and dendritic cells for antiviral and immune-based therapies. Our specific aims are: 1. To establish optimal HIV-2 gene transfer vector and stable packaging lines-Efforts will be made to minimize the chance of RCR generation, while optimizing vector titers. 2. To demonstrate protection of primary T cells, macrophages and progeny of CD34+ cells by lentiviral vector transduction against HIV challenge-HIV-2 vectors expressing anti-HIV-1 and anti-CCR5 ribozymes will be tested in these studies. We will also ascertain whether transduction by these vectors will have any deleterious effect on cell proliferation, differentiation and/or function of these target cells. 3. To examine the effects of lentiviral vector transduction on dendritic cell function and interaction with T cells- Dendritic cells (DC) are both facilitators and defenders of HIV infection since they can present both viral antigens and infectious virus to T cells. Recent data suggest that while only immature DC's are infectable by HIV, mature DC's need to express functional chemokine receptors CCR5 and CXCR4 t transmit M-tropic and T-tropic viruses to CD4+ T cells respectively. We will construct HIV-1 vectors expressing gag-pol antigens as well as anti-CC4-5 and/or anti-CXCR4 ribozymes, and determine how transduction by these vectors will impact on the ability of dendritic cells to present antigen to CD4 and CD8 cells or to transmit HIV infection to CD4 cells. In addition to these aims, this project will collaborate closely with Projects 2 and 3 to directly compare the safety and efficacy of HIV-based and FIV-based vectors and MLV-based retroviral vectors in vitro and in vivo.