Non-Integrating Lentiviral Vectors: Potential as Vaccines

非整合慢病毒载体：作为疫苗的潜力

• 批准号: 7140572
• 负责人: Mary E. Klotman
• 金额: $ 19.87万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-07-15 至 2007-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7140572
• 关键词: AIDS vaccines; antigen presenting cell; biotechnology; extrachromosomal DNA; gene expression; human immunodeficiency virus 1; human tissue; immune response; laboratory mouse; tissue /cell culture; vaccine development; vaccine evaluation; vector vaccine; virus DNA; virus integration; virus protein; virus replication; AIDS vaccines; antigen presenting cell; biotechnology; extrachromosomal DNA; gene expression; human immunodeficiency virus 1; human tissue; immune response; laboratory mouse; tissue /cell culture; vaccine development; vaccine evaluation; vector vaccine; virus DNA; virus integration; virus protein; virus replication

DESCRIPTION (provided by applicant): Despite major advancements in HIV-1 therapeutics, the HIV-1/AIDS epidemic continues to grow, highlighting the need for the development of an effective vaccine. Although an attenuated replication competent HIV-1 vaccine presents unacceptable risks, prior work with attenuated strains of SIV has led to several important observations concerning vaccine efficacy in the primate model. Chronic presentation of low levels of viral antigens in the setting of a viral infection provides protection. During retroviral infection, high levels of unintegrated extrachromosomal DNA (E-DNA) accumulate in the infected cells in addition to integrated provirus responsible for production of new viral progeny. E- DNA has been shown to persist in vivo even in the absence of detectable plasma viremia. We, as well as others, have shown that E-DNA is stable in vitro in non-dividing cells and is transcriptionally active producing functional viral proteins. However, despite the production of viral protein, E-DNA does not sustain viral replication. The central hypothesis of this proposal is that sustained viral protein production from HIV-1 E-DNA is an efficient and safe way to express viral proteins and induce an immune response. Viral proteins are presented in the context of an infectious cycle with the safety features including the lack of production of replicating virus and the absence of integration. To test this hypothesis, we will test the efficiency of integrase defective lentiviral vectors to produce viral E-DNA and proteins in professional antigen presenting cells (APC) including dendritic cells (DC). The ability of these transduced cells to be recognized by and induce a response in effector T-cells will be evaluated as well. Finally we will use a murine model to determine the kinetics of E-DNA and viral protein production from IN-defective vectors delivered intramuscularly as well as determine if the expression is associated with a measurable immune response. These feasibility studies will establish whether IN-defective vectors should be exploited for vaccine development.

描述（由申请人提供）：尽管HIV-1治疗剂取得了重大进步，但HIV-1/AIDS流行仍在增长，强调了开发有效疫苗的需求。尽管衰减的复制能干的HIV-1疫苗会出现不可接受的风险，但先前使用SIV菌株减弱的工作导致了关于灵长类动物模型中有关疫苗功效的几个重要观察结果。在病毒感染的情况下，低水平病毒抗原的长期表现可提供保护。在逆转录病毒感染期间，除了负责产生新病毒后代的综合病毒外，高水平的未集成的未集成外染色体DNA（E-DNA）积聚在感染细胞中。即使在没有可检测的血浆病毒血症的情况下，E-DN​​A也已显示出体内持续存在。我们和其他人都表明，在非分裂细胞中E-DNA在体外是稳定的，并且是转录活跃的产生功能性病毒蛋白的。但是，尽管产生了病毒蛋白，但E-DNA仍未维持病毒复制。该提议的中心假设是，HIV-1 E-DNA持续的病毒蛋白产生是表达病毒蛋白并诱导免疫反应的一种有效且安全的方法。病毒蛋白在传染性周期的背景下呈现，其安全特征，包括缺乏复制病毒的产生和缺乏整合。 为了检验该假设，我们将测试整合酶有缺陷的慢病毒载体的效率，以在包括树突状细胞（DC）（DC）的专业抗原呈递细胞（APC）中产生病毒E-DNA和蛋白质。这些转导细胞的识别和诱导效应T细胞中的反应的能力也将得到评估。最后，我们将使用鼠模型来确定肌肉内传递的缺陷载体的E-DNA和病毒蛋白产生的动力学，并确定表达是否与可测量的免疫反应有关。这些可行性研究将确定是否应利用缺陷的载体进行疫苗开发。