Development of HIV vectors for antiviral and immune based therapies

该临床前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的基于艾滋病毒的载体，包括用于抗病毒和免疫治疗的干细胞和树突状细胞。我们的具体目标是：1.建立最佳的HIV-2基因转移载体和稳定的包装线-在优化载体滴度的同时，将努力减少RCR产生的机会。2.为了证明通过慢病毒载体转导对原代T细胞、巨噬细胞和CD 34+细胞后代的保护作用，将在这些研究中检测表达抗HIV-1和抗CCR 5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.为了检查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感染的促进者又是防御者，因为它们可以将病毒抗原和感染性病毒呈递给T细胞。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需要表达功能性趋化因子受体CCR 5和CXCR 4，以分别将嗜M性和嗜T性病毒传递给CD 4 + T细胞。我们将构建表达gag-pol抗原以及抗CC 4 -5和/或抗CXCR 4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CD 4和CD 8细胞呈递抗原或将HIV感染传递给CD 4细胞的能力。除了这些目标，该项目将与项目2和3密切合作，直接比较基于HIV和FIV的载体和基于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.