Adjuvants for Agile Vaccine Development

描述（由申请人提供）：由于潜在的生物恐怖主义威胁，开发安全有效的针对选定制剂清单上的传染性微生物的疫苗是一个国家和全世界的目标。在某些情况下，已有疫苗，但它们被认为不安全，无法在大量人群中广泛使用。例如，天花减毒活疫苗和土拉杆菌减毒活疫苗仅在有限的情况下用于实验室工作人员、“第一反应者”或潜在接触风险高的其他人群。与减毒或灭活微生物疫苗相比，蛋白质亚单位疫苗和DMA疫苗是更安全的选择。最近在我们的实验室使用全蛋白质组微阵列扫描的进展导致发现了一些潜在的候选抗原用于亚单位疫苗对抗天花和土拉菌病。然而，为了提高疫苗效力和保护效果，蛋白质需要佐剂。此外，DNA疫苗的效力可以通过加入重组病毒或蛋白质来提高，“Prime-Boost”方法作为一种有效的方法来提高对重组抗原的免疫反应，正受到一些人的欢迎。然而，系统的研究旨在调查DNA引物与佐剂蛋白促进的好处是缺乏的。在这里，我们将研究在蛋白质增强中使用4种新型佐剂的DNA疫苗启动物和重组蛋白增强剂。在特异性目标1中，我们将重点使用两种模型抗原，一种来自牛痘病毒（天花疫苗），另一种来自土拉菌。野生型土拉菌基因将被优化密码子，以便在哺乳动物细胞中表达，从而使它们成为有效的DNA疫苗。每种蛋白质也将在细菌中表达，在每种佐剂中纯化和配制，以用于增强。将建立检测方法，以确定针对每种抗原的所有抗体类型和IgG亚型的血清水平，并对抗原再刺激的脾细胞进行细胞因子检测，以区分TH1和TH2反应。在特异性目标2中，所有四种佐剂将在DNA疫苗原体和蛋白质增强方案中进行评估，抗原特异性体液和细胞免疫反应将被量化。在Specific Aim 3中，将使用在我们以前的疫苗发现工作中确定的抗原组合，在针对牛痘和土拉菌病的活感染模型中测试表现最佳的疫苗配置。本申请中描述的研究的成功完成将为我们随后提交第二阶段申请的计划奠定基础，该阶段申请的重点是研究，最终将导致更安全有效的针对天花和F型螺旋体病的亚单位疫苗的临床试验。

DESCRIPTION (provided by applicant): Because of the potential bioterrorism threat, the development of safe and effective vaccines against the infectious microorganisms on the select agents list is a national and worldwide goal. In some cases there are vaccines in existence but they are considered unsafe for widespread use on large populations. For example the live attenuated smallpox and F. tularensis vaccines are used only under restricted circumstances for laboratory workers, "first responders", or for other groups at high risk of potential exposure. Protein subunit vaccines and DMA vaccines are safer alternatives compared with live attenuated or killed microorganism vaccines. Recent progress in our laboratory using whole proteome microarray scanning has lead to discovery of a number of potential candidate antigens for use in subunit vaccines against smallpox and tularemia. However, proteins require adjuvants in order to improve vaccine potency and protective efficacy. Furthermore, the potency of DNA vaccines can be increased by boosting with recombinant viruses or proteins, and the "Prime-Boost" approach is becoming accepted with some enthusiasm as an effective way to improve the immune responses against recombinant antigens. However, systematic studies designed to investigate the benefits of DNA prime together with adjuvanted protein boost are lacking. Here we will investigate the DNA vaccine prime and recombinant protein boost using 4 novel adjuvants in the protein boost. In Specific Aim 1, we will focus on using two model antigens, one derived from vaccinia virus (the smallpox vaccine) and a second from F. tularensis. The wild-type F. tularensis gene will be codon optimized for expressions in mammalian cells so that they will be effective as DNA vaccines. Each protein will also be expressed in bacteria, purified and formulated in each adjuvant for use in the boost. Assays will be established to determined serum levels of all antibody types and IgG subtypes against each antigen, and a cytokine panel will be assayed on antigen restimulated spleen cells in order to distinguish TH1 and TH2 responses. In Specific Aim 2, all four of the adjuvants will be evaluated in the DNA vaccine prime & protein boost protocol, and antigen specific humoral and cellular immune responses will be quantified. In Specific Aim 3 the best performing vaccine configurations will be tested in live infection models against vaccinia and F. tularensis using combinations of antigens identified in our previous vaccine discovery effort. Successful completion of the studies described in this application will set a stage for our subsequent plan to submit a Phase 2 application focusing on studies that will eventually lead to clinical trials of safer and effective subunit vaccines against smallpox and F tularensis.