Bacteriophage gene transfer for HIV vaccine delivery

用于 HIV 疫苗递送的噬菌体基因转移

基本信息

批准号：
7002647
负责人：
Stephen Dewhurst
金额：
$ 22.85万
依托单位：
UNIVERSITY OF ROCHESTER
依托单位国家：
美国
项目类别：
财政年份：
2005
资助国家：
美国
起止时间：
2005-01-01 至 2007-09-30
项目状态：
已结题

项目摘要

DESCRIPTION (provided by applicant): Several mammalian virus vectors are in human clinical trial to evaluate their potential as HIV vaccine delivery platforms, but these vectors also have significant drawbacks that include safety concerns, high production and purification costs, and a host cell tropism that is often very broad. In contrast,bacteriophage vectors are cheap to manufacture and purify, and can be selectively targeted to mammalian cells through engineered modifications of the phage coat proteins. The central hypothesis of this Innovation grant application is therefore as follows: (i) that bacteriophage vectors can be retargeted so as to allow efficient transduction of dendritic cells (DC), and (ii) that these modified bacteriophage vectors can be used to elicit specific and potent immune responses to an encoded (HIV-1) antigen. This hypothesis will be tested experimentally, in a proof-of-principle study, through three specific aims. In the first aim, modified bacteriophage vectors capable of transducing DC will be generated, using multiple complementary approaches. To do this, a mammalian GFP reporter cassette will be inserted into our phage vectors, and these GFP-phage will then be subjected to specific coat protein modifications intended to enhance phage uptake into DC; a series of complementary approaches that will be evaluated in parallel in order to determine the optimum approach. In the second specific aim, an HIV-1 gene insert will be introduced into the genomes of the DC-targeted phage vectors, and their ability to express HIV-1 antigens in DC will be evaluated. Finally, in Aim 3, the most promising HIV-1 antigen-encoding phage vectors will be inoculated into BALB/c mice and into guinea pigs, together with relevant control phage, and their ability to elicit HIV-1-specific cellular and humoral immune responses will be examined. These experiments will also include studies to determine the effectiveness of multiple immunizations with phage vectors (i.e., to test whether anti-phage antibodies induced after an initial immunization have an impact upon subsequent boosting with the same phage, or different/modified phage).

描述（由申请人提供）：几种哺乳动物病毒载体正在进行人体临床试验，以评估其作为 HIV 疫苗递送平台的潜力，但这些载体也具有显着的缺点，包括安全问题、高生产和纯化成本以及宿主细胞向性往往非常宽泛。相比之下，噬菌体载体的制造和纯化成本低廉，并且可以通过噬菌体外壳蛋白的工程修饰选择性地靶向哺乳动物细胞。因此，本创新拨款申请的中心假设如下：（i）噬菌体载体可以重新定位，以便有效转导树突状细胞（DC），以及（ii）这些修饰的噬菌体载体可用于引发特定的噬菌体载体。对编码的 (HIV-1) 抗原产生有效的免疫反应。这一假设将在原理验证研究中通过三个具体目标进行实验检验。在第一个目标中，将使用多种互补方法产生能够转导 DC 的修饰噬菌体载体。为此，将哺乳动物 GFP 报告基因盒插入我们的噬菌体载体中，然后对这些 GFP 噬菌体进行特定的外壳蛋白修饰，旨在增强噬菌体对 DC 的摄取；一系列互补的方法将被并行评估以确定最佳方法。在第二个具体目标中，将 HIV-1 基因插入片段引入 DC 靶向噬菌体载体的基因组中，并评估它们在 DC 中表达 HIV-1 抗原的能力。最后，在目标 3 中，最有前途的 HIV-1 抗原编码噬菌体载体将与相关对照噬菌体一起接种到 BALB/c 小鼠和豚鼠中，并具有引发 HIV-1 特异性细胞和体液免疫的能力。将对答复进行审查。这些实验还将包括确定噬菌体载体多次免疫有效性的研究（即测试初次免疫后诱导的抗噬菌体抗体是否对随后使用相同噬菌体或不同/修饰噬菌体的加强有影响）。