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，针对选定制剂名单上的传染病微生物开发安全有效的疫苗是一个国家和世界目标。在某些情况下，存在疫苗，但它们被认为在大量人口上广泛使用是不安全的。例如，天花减毒活疫苗和图拉氏丝虫减毒活疫苗仅在实验室工作人员、“急救人员”或潜在接触风险较高的其他群体的受限情况下使用。与减毒或灭活微生物疫苗相比，蛋白质亚单位疫苗和DMA疫苗是更安全的替代品。我们实验室使用全蛋白质组微阵列扫描的最新进展导致发现了一些潜在的候选抗原，可用于天花和兔热病的亚单位疫苗。然而，蛋白质需要佐剂才能提高疫苗效力和保护效力。此外，DNA疫苗的效力可以通过重组病毒或蛋白质的增强来增加，而“Prime-Boost”方法正被一些人热情地接受为提高对重组抗原的免疫应答的有效方法。然而，缺乏系统的研究来研究DNA PRIME和辅助性蛋白Boost的益处。在这里，我们将研究在DNA疫苗中使用4种新型佐剂的重组蛋白Boost和DNA疫苗。在具体目标1中，我们将重点使用两种模式抗原，一种来自痘苗病毒(天花疫苗)，另一种来自图拉氏丝虫。野生型图拉氏丝虫基因将对密码子进行优化，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，纯化并在每种佐剂中配制，用于加强免疫。将建立针对每个抗原的所有抗体类型和Ig G亚型的血清水平的检测方法，并将对抗原重新刺激的脾细胞进行细胞因子小组检测，以区分TH1和TH2反应。在特定目标2中，所有四种佐剂都将在DNA疫苗初级和蛋白质增强方案中进行评估，并将量化抗原特异性体液和细胞免疫反应。在具体目标3中，将使用我们以前的疫苗发现工作中确定的抗原组合，在针对牛痘和图拉氏丝虫的活体感染模型中测试性能最好的疫苗配置。本申请中描述的研究的成功完成将为我们后续计划提交第二阶段申请奠定基础，重点研究最终将导致更安全和有效的亚单位疫苗的临床试验，以对抗天花和图拉氏菌。

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.