Feline immunodeficiency vector system

猫免疫缺陷载体系统

• 批准号: 6314014
• 负责人: David James Looney
• 金额: $ 20.81万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-08-15 至 2000-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6314014
• 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为人类细胞提供了潜在的更安全和更方便的基因转移。虽然灵长类慢病毒来自致命的人类病原体，但没有证据表明人类可以感染FIV。在艾滋病毒感染者中动员艾滋病毒媒介，虽然可能是一种优势，但也引起了将媒介转移到包括生殖细胞在内的非预期目标的担忧。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。使用异源启动子可以在人类细胞中生产基于FIV的载体(避免了非人类细胞中存在的内源性逆转录病毒的污染风险，同时保留了理想的Lenti逆转录病毒转导未分裂的、终末分化的细胞，如神经元和单核巨噬细胞的能力。然而，基于FIV和HIV的载体系统的转导和表达的相对效率仍有待确定。FIV包装系统通过使用单独的结构将重组降到最低，目前还没有开发出来。表达核酶的FIV载体的抗病毒活性尚未被检测，也没有研究HIV或SIV感染对FIV载体的动员作用。本项目的主要目标是：[1]比较类似的FIV和基于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]建立安全高效的包装细胞系，用于生产高效的FIV载体，[3]比较HIV和HIV表达抗HIV或SIV核酶的效率，以及[4]检测FIV载体转导能够在猕猴模型中再繁殖的灵长类造血干细胞的能力，该项目应提供所需的宝贵信息，以便合理选择FIV或艾滋病毒载体，用于临床应用，如艾滋病毒感染的基因治疗。