POLYNUCLEOTIDE VACCINES FOR SIV INFECTION AND DISEASE

本申请建议独立调查员支持候选人 自2001年以来，他一直积极参与非病毒基因传递的开发， 1986年，参加了一些原创和重要的研究 包括导致多核苷酸疫苗概念的研究。 范围具体涉及提供资金支持 对主要研究者进行为期五年的培训。 的 大部分工作都得到了一个积极的ARMDC奖项的支持，该奖项将包括 调查的所有其他费用。 最近，候选人成立了 并指导了一个小型的学术基因治疗项目， 用于粘膜基因递送的方法和多核苷酸疫苗。 该基因 一个治疗计划已经在加州大学组装， 戴维斯，在那里它已经能够利用大量的专业知识， 加州大学戴维斯分校CFAR研究社区和当地灵长类动物研究中心 （CRFRC）。 候选人目前持有兼职任命， 所要求的支助将使工作人员的薪金有一定的稳定性， 候选人，允许进一步发展作为一个独立的学术 调查员 候选人打算继续把重点放在 开发和测试非病毒基因转移技术， 将这些技术应用于适当的临床目标。 的 候选人认为多核苷酸疫苗是最合适的， 目前非病毒基因递送技术的治疗应用， 因此，重点是开发这种疫苗， 慢病毒性疾病和急性病毒性肺炎。 基因免疫（DNA 疫苗接种）和抗病毒免疫应答的细胞因子增强， 疫苗研究的新领域。 有限的研究表明， 表达病毒基因的质粒诱导保护性抗病毒免疫 直接注射后的反应，以及外源性给药 细胞因子也可以影响预先存在或发展中的宿主免疫 应答 然而，基因疫苗的功效（有或没有）， 细胞因子和/或B7），所述细胞因子和/或B7）针对引起致死性 灵长类动物（包括人类）的免疫缺陷尚未得到证实。 在 此外，我们还没有阐明接种疫苗的灵长类动物的免疫反应。 能提供保护的和不能提供保护的。 该提案旨在 在定义一个“安全的”遗传免疫协议，能够诱导 对SIV感染和疾病的保护性免疫，同时 确定负责这种保护性免疫的免疫机制。 这些机制的确定可望进一步开辟道路， 有效的暴露后免疫疗法， 个体 此应用程序将为实现 具体目标; 1. 慢病毒基因增强表达载体的研制 proteins. 2. 产生和评估对慢病毒蛋白的免疫应答， 使用小鼠和猕猴模型直接DNA注射。 3. 产生和评估对慢病毒蛋白的免疫应答， 直接DNA注射表达慢病毒蛋白和 细胞因子 4. 评价已接种疫苗的猕猴的临床病程 使用DNA注射并用致病性SIV攻击。

This application proposes independent investigator support for a candidate who has been active in the development of non-viral gene delivery since 1986, and has participated in a number of original and significant studies including studies resulting in the conception of polynucleotide vaccines. The scope specifically involves the provision of funds to support the training of the principal investigator over a five year period. The majority of the work is supported by an active ARMDC award which will cover all other costs of the investigation. Recently the candidate has founded and directs a small academic gene therapy program, which has developed methods for mucosal gene delivery and polynucleotide vaccines. This gene therapy program has been assembled at the University of California at Davis, where it has been able to exploit the substantial expertise of the UC Davis CFAR research community and the local primate research center (CRFRC). The candidate currently holds an adjunct appointment, and the requested support would provide for some stability in salary for the candidate, allowing further development as an independent academic investigator. The candidate intends to continue to focus on the development and testing of non-viral gene transfer technologies and the application of those technologies to appropriate clinical targets. The candidate believes that polynucleotide vaccines are ht most appropriate therapeutic application for current non-viral gene delivery technologies, and hence is focused on the development of such vaccines for prevention of lentiviral disease and acute viral pneumonia. Genetic immunization (DNA vaccination) and cytokine augmentation of antiviral immune responses are novel areas of vaccine research. Limited studies have demonstrated that plasmids expressing viral genes induce protective antiviral immune responses after direct injection, and that exogenously administered cytokines can also influence pre-existing or developing host immune responses. However, the efficacy of genetic vaccines (with or without cytokines and /or B7) directed against lentiviruses which cause fatal immunodeficiency in primates (including humans) has not been proven. In addition, wee have yet to clarify immune responses in vaccinated primates that do confer protection versus those that do not. This proposal is aimed at defining a "safe" genetic immunization protocol capable of inducing protective immunity to SIV infection and disease, and concurrently identifying the immune mechanisms responsible for this protective immunity. The identification of these mechanisms is expected to further open the way to effective post exposure immunotherapies for already infected individuals. This application will provide support for achieving the following specific aims; 1. Development of DNA vectors for enhanced expression of lentiviral proteins. 2. Generating and evaluating immune responses to lentiviral proteins after direct DNA injection using mouse and macaque models. 3. Generating and evaluating immune responses to lentiviral proteins after direct DNA injection of vectors expressing both lentiviral proteins and cytokines. 4. Evaluation of clinical course of macaques which have been vaccinated using DNA injection and challenged with pathogenic SIV.