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，开发针对特定病原体清单上的传染性微生物的安全有效的疫苗是国家和全球的目标。在某些情况下，疫苗是存在的，但它们被认为是不安全的，不能广泛用于大规模人群。例如活的减毒天花和F.土拉热疫苗只在有限的情况下用于实验室工作人员、“第一反应者”或其他可能接触的高风险群体。与减毒活疫苗或灭活微生物疫苗相比，蛋白亚单位疫苗和DMA疫苗是更安全的替代品。最近在我们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天花和兔热病。然而，蛋白质需要佐剂以提高疫苗效力和保护效力。此外，DNA疫苗的效力可以通过用重组病毒或蛋白质加强来增加，并且“初免-加强”方法正被一些热情地接受为改善针对重组抗原的免疫应答的有效方式。然而，缺乏旨在研究DNA初免与佐剂蛋白加强的益处的系统研究。在这里，我们将研究DNA疫苗初免和重组蛋白加强免疫，在蛋白加强免疫中使用4种新型佐剂。在具体目标1中，我们将集中使用两种模型抗原，一种来自牛痘病毒（天花疫苗），另一种来自F。土拉热。野生型F.土拉热菌基因将被密码子优化以在哺乳动物细胞中表达，从而它们将作为DNA疫苗有效。每种蛋白质也将在细菌中表达，纯化并配制在每种佐剂中用于加强。将建立测定法，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，并将在抗原再刺激的脾细胞上测定细胞因子组，以区分TH 1和TH 2应答。在特定目标2中，将在DNA疫苗初免和蛋白质加强方案中评估所有四种佐剂，并将量化抗原特异性体液和细胞免疫应答。在特定目标3中，将在抗牛痘和F.使用在我们以前的疫苗发现工作中鉴定的抗原的组合来治疗土拉热。本申请中描述的研究的成功完成将为我们随后提交II期申请的计划奠定基础，该计划将重点关注最终将导致针对天花和土拉杆菌的更安全有效的亚单位疫苗的临床试验的研究。

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.