Feline immunodeficiency vector system

猫免疫缺陷载体系统

• 批准号: 6484681
• 负责人: David James Looney
• 金额: $ 24.75万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-08-01 至 2002-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6484681
• 关键词: HIV infections; biotechnology; clinical research; feline immunodeficiency virus; gene therapy; genetic promoter element; genetic transduction; hematopoietic stem cells; human immunodeficiency virus; human subject; human tissue; leukapheresis; phlebotomy; ribozymes; technology /technique development; tissue /cell culture; transfection /expression vector; virus infection mechanism

Non-primate lentiviral vectors offer potentially safer and more convenient gene transfer to human cells than vectors based upon HIV-1. While the primate lentiviruses are derived from lethal human pathogens, there is no evidence that humans can be infected by FIV. Mobilization of HIV vectors in HIV infected individuals, while a possible advantage, raises the concern of transfer of vector to unintended targets, including germ cells. Recently, our studies showed that the only restriction to productive infection of human cells by feline immunodeficiency virus (FIV) is the inactivity of the FIV promoter. The use of a heterologous promoter allows production of FIV-based vectors in human cells (avoiding the risk of contamination by endogenous retroviruses present in non-human cells, while retaining the desirable lenti-retroviral capacity to transduce non-dividing, terminally differentiated cells such as neurons and monocyte-macrophage. However, the relative efficiency of FIV- and HIV-based vector systems for transduction and expression remains to be determined. FIV packaging systems which minimize recombination by use of separate constructs have yet to be developed. The antiviral activity of FIV vectors expressing ribozymes has not been examined, and the mobilization of FIV vectors by HIV or SIV infection has not been studied. The major goals of this project are: [1] To compare transduction efficiency of analogous FIV and HIV based vectors, and the extent to which these vectors are mobilized by HIV or SIV infection in vitro, [2] To develop safe and efficient packaging cell lines for production of high-titer FIV vectors, [3] To compare the efficiency of HIV and HIV expressing anti- HIV or SIV ribozymes to confer antiviral resistance, and [4] To examine the ability of FIV vectors to transduce primate hematopoietic stem cells capable of repopulation in macaque models, in collaboration with Project III. This project should provide valuable information needed to make a rational choice of FIV or HIV vectors for clinical applications such as gene therapy for HIV infection.

与基于HIV-1的载体相比，非灵长类慢病毒载体提供了潜在的更安全和更方便的向人类细胞的基因转移。虽然灵长类动物慢病毒来源于致命的人类病原体，但没有证据表明人类可以被FIV感染。在HIV感染个体中HIV载体的动员虽然是一个可能的优势，但引起了对载体转移到非预期目标（包括生殖细胞）的关注。最近，我们的研究表明，猫免疫缺陷病毒（FIV）对人类细胞的生产性感染的唯一限制是FIV启动子的失活。异源启动子的使用允许在人细胞中产生基于FIV的载体（避免了被非人细胞中存在的内源性逆转录病毒污染的风险，同时保留了期望的慢病毒-逆转录病毒能力以使非分裂的终末分化细胞如神经元和单核细胞-巨噬细胞增殖）。然而，基于FIV和HIV的载体系统用于转导和表达的相对效率仍有待确定。通过使用单独的构建体使重组最小化的FIV包装系统还有待开发。表达核酶的FIV载体的抗病毒活性尚未被检测，并且尚未研究HIV或SIV感染对FIV载体的动员。该项目的主要目标是：[1]为了比较类似的基于FIV和HIV的载体的转导效率，以及这些载体在体外被HIV或SIV感染动员的程度，[2]为了开发用于生产高滴度FIV载体的安全和有效的包装细胞系，[3]为了比较HIV和表达抗HIV或SIV核酶的HIV赋予抗病毒抗性的效率，和[4]与项目III合作，检查FIV载体在猕猴模型中复制能够再增殖的灵长类造血干细胞的能力。该项目应提供有价值的信息，需要作出合理的选择FIV或HIV载体的临床应用，如基因治疗艾滋病毒感染。