RNA DELIVERY FOR DENDRITIC CELL HIV ANTIGEN PREVENTION

提供了 一种安全有效的艾滋病疫苗是抗击艾滋病全球蔓延的关键。Whilethe 免疫保护的相关性尚未明确定义，无论是保护性疫苗还是治疗性疫苗， 认为CD 4 ~* 和CD 8细胞以及体液免疫都是重要的。多么广泛， 有效的，可持续的反应可以引起尚未确定，这代表了我们的关键差距， 了解如何产生一种有效的疫苗，从而实现保护性免疫。树突状细胞 (DC)代表了先天免疫和适应性免疫之间的界面，是最有潜力和最重要的 因此，我们专注于新的方法来操纵DC 以利用它们在抗HIV免疫应答中的潜在作用。一个最近开发的，特别是 有希望的引发DC的方法是使用抗原编码的mRNA的转染。为此，我们开发了 一种新的，高效的方法负载DC与mRNA，并利用这种方法来最大限度地提高DC免疫 反应生成能力。在我们的初步研究中，我们已经表明，编码抗原负载DC的mRNA 能够引发CD 4和CD 8两种应答。此外，抗原编码的mRNA可以直接被导入细胞内。 小鼠并产生初级和次级T和B细胞免疫应答。此外，我们的试点数据表明， 除了提供抗原递送模式之外，RNA还具有独特的激活DC的内在能力。我们 假设是基于RNA的树突状细胞抗原递送提供了独特的灵活和有效的方法 对于免疫，RNA传递的机制可以被操纵来调节反应的类型， 产生（包括辅助，细胞毒性和体液免疫），这种方法可以用于 艾滋病疫苗的研制。该提案的目的是更好地了解RNA 编码的抗原被DC加工，定义了特异性抗原呈递途径可能被DC加工的因素。 通过RNA编码的抗原靶向，并建立对HIV有效性的概念验证证据。我们预计， 这项工作将增强我们对DC免疫功能的理解，使我们能够差异化地操纵特定的元件， 在免疫反应中，并最终使我们能够利用mRNA编码的抗原负载树突状细胞产生的 免疫反应的潜在保护和治疗目的，除了提供一个基础， 一个理性的发展。在艾滋病毒疫苗开发方面，我们希望这些研究也能提供 a new新avenue大道for vaccine疫苗development发展in general一般.

PROVIDED. A safe andeffectivevaccinefor HIV is criticallyneededto combatthe worldwidescourgeof AIDS. Whilethe correlatesof immuneprotectionhave yet to be clearlydefined,eitherforprotectiveor therapeuticvaccines,it is widely believed that both CD4* and CD8 cell as well as humoralimmunityare all important. How sufficientlybroad, potent,and sustainedresponsescan be elicitedhas yet to be determined,and this representsa criticalgap in our understandingof howto generatean effectivevaccinesuchthat protectiveimmunitycan be achieved. Dendriticcells (DC) representthe interfacebetween innate and adaptiveimmunityand are amongthe most potentand important componentsin generatingimmune responses.Therefore,we havefocusedon novelapproachesto manipulatingDC in order to exploit their potential role in anti-HIV immune responses. One recently developed and particularly promising approach to priming DC is using transfection of antigen encoded mRNA. Towards this end, we developed a novel, high-efficiency method of loading DC with mRNA, and utilized this approach to maximize DC immune response generation capacity. In our preliminary studies, we have shown that mRNA encoding antigen-loaded DC are able to elicit both CD4 CD8 responses. In addition, antigen-encoded mRNA can be directly iniected into mice and generate both primary and secondary T and B cell immune responses. Furthermore, our pilot data suggest that in addition to providing a mode of antigen delivery, RNA has a unique intrinsic ability to activate DC. Our hypotheseis is that RNA-based dendritic cell antigen delivery offers a uniquely flexible and potent approach to immunization, that the mechanism of RNA delivery can be manipulated to regulate the types of response generated (including helper, cytotoxic and humoral immunity), and that this approach can be exploited for the development of HIV vaccines. The aim of this proposal is to better understand the mechanisms by which RNA encoded antigen is processed by DC, define the factors by which specific antigen presentation pathways may be targeted by RNA-encoded antigen, and establish proof-of-concept evidence for efficacy in HIV. We anticipate that this work will enhance our understanding of DC immune function, enable us to differentially manipulate specific elements in the immune response, and, ultimately, allow us to exploit mRNA encoded antigen loaded dendritic cell-generated immune responses for potentially protective and therapeutic purposes, in addition to providing a foundation for the rational development of a now._lapproach to HIV vaccine development, it is our hope that these studies may also offer a new avenuefor vaccine development in general.