Rapid deployment DNA vaccine for pandemic influenza

描述（由申请人提供）：仅接种DNA疫苗将大大提高DNA疫苗在大流行应用中的快速部署效用（DNA疫苗的开发时间明显短于蛋白质或病毒载体系统的开发时间）。该项目的总体目标是开发一种可预防H5N1大流行性流感的快速部署DNA疫苗平台。假设流感DNA疫苗的效力可以通过载体编码的免疫刺激RNA激活细胞质双链RNA （dsRNA）信号通路来提高。这将通过使用各种免疫刺激“RNA元件”（RNAe）来实现。这项I期概念验证研究将调查包含载体介导的RNAe对载体编码靶抗原免疫应答的潜在益处。流感血凝素（HA）将作为需要抗体反应保护的抗原的例子。产生免疫刺激性dsRNA的新型HA DNA疫苗质粒载体将被创造出来，并通过电穿孔增强肌肉注射给药后在体内筛选免疫反应。将选择改善中和抗体对HA反应的最佳骨架。然后，在这个RNAe载体的基础上，“抑制抑制剂”的DNA疫苗将被创造出来，并在从属研究中进行测试。第二种假设是，dsRNA激活DNA疫苗可通过阻止靶细胞下调载体激活先天免疫介导的免疫反应，进一步改善dsRNA激活DNA疫苗诱导的免疫反应。将进一步修改上述定义的载体，以纳入RNAe，该RNAe可敲除这些细胞质dsRNA信号通路的各种衰减因子的表达。随后将在动物模型中确定“抑制-抑制”DNA疫苗对HA诱导的抗体反应。在第二阶段，将开发出最佳载体骨架（即单独使用特异性免疫刺激RNAe，或可能与本文确定的“抑制抑制剂”shRNA联合使用），作为一种新型的、快速部署的H5N1流感单dna疫苗平台。本文开发的快速部署DNA疫苗平台将与各种其他新兴生物制剂具有普遍效用；NTC将授权该平台进行此类应用。这一建议旨在开发新型流感疫苗，以取代基于鸡蛋的疫苗生产技术，因此符合非ai -06-011中概述的NIAID的高优先流感研究领域。

DESCRIPTION (provided by applicant): DNA-only vaccination would significantly enhance the rapid deployment utility of DNA vaccines for pandemic application (development times for DNA vaccines are significantly shorter than those for protein or viral vector systems). The overall goal of this project is to develop a rapid deployment DNA vaccine platform protective against pandemic influenza H5N1. The hypothesis is that influenza DNA vaccine potency can be increased through activation of cytoplasmic double stranded RNA (dsRNA) signaling pathways by vector encoded immunostimulatory RNA. This will be accomplished through the use of various immunostimulatory "RNA elements" (RNAe). This Phase I proof of concept study will investigate the potential benefit of inclusion of vector-mediated RNAe on immune responses to an vector encoded target antigen. Influenza Hemagglutinin (HA) will be used as an example antigen requiring antibody responses for protection. Novel HA DNA vaccine plasmid vectors that produce immunostimulatory dsRNA will be created and screened for immune response in vivo after delivery by electroporation-enhanced intramuscular injection. The optimal backbone that improves neutralizing antibody response to HA will be selected. Then, building upon this RNAe vector, 'inhibiting-the-inhibitor' DNA vaccines will be created and tested in a subordinate study. The secondary hypothesis is that immune responses induced by the dsRNA activating DNA vaccines can be further improved by preventing target cell downregulation of immune responses mediated by vector activation of innate immunity. The vectors defined above will be further modified to incorporate RNAe which knockdown expression of various attenuators of these cytoplasmic dsRNA signaling pathways. Antibody responses to HA induced by the 'inhibiting-the- inhibitor' DNA vaccines will then be determined in an animal model. In Phase II the resulting optimal vector backbone (i.e., specific immunostimulatory RNAe alone, or potentially in combination with 'inhibiting the inhibitor' shRNA as determined herein) will be developed as a novel, rapid deployment DNA-only vaccine platform for influenza H5N1. The rapid deployment DNA vaccine platform developed herein will have general utility with a variety of other emerging biological agents; NTC will license the platform for such applications. This proposal is aimed at developing novel influenza vaccines to replace egg based vaccine production technologies and as such is responsive to NIAID's high priority influenza research areas, as outlined in NOT-AI-06-011.