FOAMY VIRUS VECTOR FOR IMMUNODEFICIENCY VIRUS THERAPY

用于免疫缺陷病毒治疗的泡沫病毒载体

• 批准号: 6020016
• 负责人: AYALEW MERGIA
• 金额: $ 16万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-09-30 至 2003-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6020016
• 关键词: AIDS therapy; Retroviridae; SCID mouse; disease /disorder model; gene therapy; simian AIDSs; simian immunodeficiency virus; technology /technique development; transfection /expression vector; vaccine development; vector vaccine; viral vaccines

Current available retroviral vectors for gene therapy are based on murine and avian retroviruses. The efficiency gene transfers in primate primary cells by these vectors is very low and functional vectors are eventually lost. Therefore, novel gene therapy approaches to combat HIV infection will be limited as a result of inefficient delivery system. Since primate retroviruses replicate efficiently in primate cells a primate based retroviral vector will be a better system for antiviral therapy against HIV infection. Primate foamy viruses have several inherent features that make them ideal for gene transfer in primates. The foamy viruses are members of the spumaviriniae sub-family of retroviruses. Foamy viruses are found in many mammalian species including primates. These viruses appear to be non-pathogenic in their natural host even though virus is widely distributed in an animal and has been recovered from several organs and tissues including brain and peripheral blood leukocytes. Also, foamy viruses have a broad host range in cell culture with respect to cell type and species and can be propagated in cells such as epithelial and fibroblast cells as well as lymphoid cells and neural cells. A foamy virus isolate from one species can also infect other mammalian species. Thus, foamy viruses offer unique opportunities for developing viral vector systems to deliver genes into several cell types of many species. Recently, the genome of simian foamy virus type l (SFV-l) from rhesus macaque has been molecularly characterized by the Principal Investigator. The goal of this proposal is to develop a retroviral vector based on SFV- l for antiviral therapy against simian immunodeficiency virus (SIV) infection. SPECIFIC AIM 1: Packaging cell lines will be established, and SFV-l vectors with a selectable marker (neomycin) will be constructed. SPECIFIC AIM 2: The efficacy of SFV-l vectors will be tested in primate tissue culture cell lines and primary cells with emphasis on bone marrow cells. SPECIFIC AIM 3: SFV-l vector containing multiple ribozymes for SIV will be constructed and the antiviral activity of the multiple ribozymes will be tested. Ribozymes with multiple targets in the viral genome are expected to be effective inhibitors of SIV replication. Experimental outcomes from these studies will help device ribozyme based therapy with efficient vector system that prevent HIV infection.

目前可用于基因治疗的逆转录病毒载体是基于小鼠的 和禽类逆转录病毒。灵长类初级动物基因转移的效率 细胞通过这些载体是非常低的，功能载体最终是 迷路了。因此，对抗HIV感染的新的基因治疗方法 由于交付系统效率低下，将受到限制。自灵长类以来 逆转录病毒在以灵长类为基础的灵长类细胞中高效复制 逆转录病毒载体将成为抗病毒治疗的更好系统 艾滋病毒感染。灵长类泡沫病毒有几个固有的特征 使它们成为灵长类动物基因转移的理想选择。泡沫病毒是 海绵病毒亚科逆转录病毒的成员。泡沫病毒是 存在于包括灵长类在内的许多哺乳动物物种中。这些病毒出现在 在它们的自然宿主中是非致病的，即使病毒广泛存在 分布于动物体内，已从几个器官和 组织包括脑和外周血白细胞。还有，起泡的 就细胞类型而言，病毒在细胞培养中具有广泛的宿主范围 和物种，并可在上皮细胞和 成纤维细胞以及淋巴样细胞和神经细胞。一种泡沫病毒 从一个物种分离出来的病毒也可以感染其他哺乳动物物种。因此， 泡沫病毒为发展病毒载体提供了独特的机会 将基因输送到许多物种的几种细胞类型中的系统。 最近，从恒河猴分离到的L泡沫病毒的基因组 首席调查员对猕猴进行了分子特征研究。 这项提议的目标是开发基于SFV的逆转录病毒载体- L在猴免疫缺陷病毒的抗病毒治疗中 感染。 具体目标1：将建立包装细胞系，并将SFV-L 将构建带有可选择标记(新霉素)的载体。 特异性目的2：在灵长类动物中测试SFV-L载体的效果 以骨髓为重点的组织培养细胞系和原代细胞 细胞。 目的构建含多种核酶的猪瘟病毒L载体 构建了多种核酶，并将其抗病毒活性 测试过。 在病毒基因组中具有多个靶点的核酶有望成为 有效的SIV复制抑制剂。这些实验的结果 研究将有助于利用有效载体进行基于核酶的治疗 预防艾滋病毒感染的系统。