Novel Smallpox Vaccine Derived from VV/VAR Immunome

描述（由申请人提供）：EpiVax专门通过筛选候选疫苗组分的全基因组来开发表位驱动的疫苗。在本申请中，我们建议开发一种安全的新型天花疫苗，该疫苗基于牛痘病毒（VV）和天花病毒（VAR）“免疫组”之间保守的表位，可在发生生物恐怖袭击时用作预防性和治疗性干预。首先，通过计算机驱动的（EpiMatrix）分析筛选在牛痘病毒（W）和天花病毒（Var）基因组之间保守的肽序列的免疫显性表位，并使用来自VV免疫个体的T细胞进行确认。第二，然后将选定的表位进行比对并克隆到DNA疫苗载体中，所述DNA疫苗载体被优化用于体内强烈和持续的表达。第三，我们将用DNA疫苗构建体接种HLA转基因小鼠，使用细胞内细胞因子流式细胞术测量对DNA疫苗构建体的从头免疫应答。EpiVax将负责VV/Var表位的选择和项目管理。布朗大学的TB/HIV研究实验室（分包商）将进行结合试验，并开发由选定表位串组成的寡核苷酸多表位构建体。Crowe博士的实验室（分包商，范德比尔特大学）将进行ELISpot试验。Dr. Weiner（分包商，Stellar Chance Laboratories，宾夕法尼亚大学）将进行疫苗接种研究。第一阶段的最终结果将是对几种原型天花疫苗构建体在HLA转基因小鼠中的免疫原性和保护作用的评价。在第二阶段，我们将筛选额外的表位，构建最终的疫苗构建体，用佐剂和递送载体优化最终疫苗构建体的免疫原性，并解决构建体在人类受试者中的安全性、毒性和免疫原性。

DESCRIPTION (provided by applicant): EpiVax specializes in the development of epitope-driven vaccines by screening whole genomes for candidate vaccine components. In this application, we propose to develop a safe, new smallpox vaccine based on epitopes conserved between the vaccinia virus (VV) and Variola (VAR) 'immunomes' that could be used as both a prophylactic and a therapeutic intervention in the event of a bioterrorist attack. First, peptide sequences that are conserved between the vaccinia virus (W) and Variola (Var) genomes will be screened for immunodominant epitopes by computer-driven (EpiMatrix) analysis and confirmed using T cells from VV-immunized individuals. Second, selected epitopes will then be aligned and cloned into DNA vaccine vectors optimized for strong and sustained expression in vivo. Third, we will vaccinate HLA-transgenic mice with the DNA vaccine constructs, measuring de novo immune responses to the DNA vaccine constructs using intracellular cytokine flow cytometry. EpiVax will be responsible for selecting the VV/Var epitopes and managing the project. The TB/HIV Research Lab at Brown University (subcontractor) will perform binding assays and develop the oligonucleotide multi-epitope constructs consisting of strings of the selected epitopes. Dr. Crowe's laboratory (subcontractor, Vanderbilt University) will perform ELISpot assays. Dr. Weiner (subcontractor, Stellar Chance Laboratories, University of Pennsylvania) will perform the vaccination studies. The final product of Phase I will be an evaluation of the immunogenicity and protective effect of several prototype smallpox vaccine constructs in HLA transgenic mice. In Phase II we would screen additional epitopes, build the final vaccine constructs, optimize the immunogenicity of the final vaccine constructs with adjuvants and delivery vehicles, and address the safety, toxicity and immunogenicity of the constructs in human subjects.