DEVELOPMENT OF A VIRUS-FREE DNA VACCINE AGAINST SMALLPOX

开发针对天花的无病毒 DNA 疫苗

• 批准号: 8360153
• 负责人: Miguel Otero
• 金额: $ 10.26万
• 依托单位: UNIVERSITY OF PUERTO RICO
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-06-01 至 2012-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8360153
• 关键词: Adjuvant; Animals; Antigens; Biomedical Research; Categories; Cells; Core Protein; DNA Vaccines; Dendritic Cells; Development; Dose; Elderly; Enzyme-Linked Immunosorbent Assay; Epitopes; Event; Funding; Genes; Grant; Growth Factor; Histocompatibility Antigens Class II; Immune; Immune response; Immunocompromised Host; Immunology; Life; Macrophage Inflammatory Protein-1; Maps; Mediating; Memory; Modeling; Molecular; Mus; National Center for Research Resources; Population; Pregnant Women; Principal Investigator; Puerto Rico; Research; Research Infrastructure; Resources; Smallpox; Source; Structural Protein; System; Testing; Transplant Recipients; United States National Institutes of Health; Vaccinated; Vaccination; Vaccines; Vaccinia virus; Viral Load result; Viral Proteins; Virus; chemokine; cost; enzyme linked immunospot assay; hazard; interest; pathogen; plasmid DNA

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Smallpox is a threat in an event of a bioterrorist attack, therefore it is important to develop a safe vaccine against this virus. There is an important population of non-vaccinated immunocompromised people that could not be safely vaccinated with the current vaccine. Within these are patients of transplants, under immunosupressive therapy, pregnant women, and the elderly. We are interested in the development of a safer vaccine that protects the population against a possible attack. We hypothesize that the adjuvant-mediated enhancement of the molecular antigen immune response will control the viral burden on infected mice. However, as smallpox is only one of possible threats, this project will provide an excellent model to test the hypothesis of a molecular adjuvant that will help set up a system that could be applied to other Category A, B and C Priority Pathogens. Mice will be immunized with plasmid DNA encoding for the vaccinia virus proteins C7L and A55R genes that are non structural proteins known to generate a cellular immune response, and the recently discovered E6R and L3L core proteins. The immunology of those molecules has not been tested to the best of our knowledge. Animals will be vaccinated with each construct individually or combined with two molecular adjuvants, the chemokine MIP-1-alpha and another construct which encodes for the dendritic cell-specific growth factor Flt-3L, a combination that is known to increase the recruitment and expansion of dendritic cells. To the best of our knowledge, this combination of molecular adjuvants has never been tested in the context of smallpox vaccination. Cell- and humoral-mediated immune responses from each antigen will be determined by ELISPOT and ELISA, respectively. Molecular mapping of each antigen will be performed to identify dominant epitopes. Animals will be challenged with a lethal dose of vaccinia virus, and the viral load will be determined by qPCR analysis. Memory, activation and proliferation will be determined to detect the long-lasting effect of each antigen on the induction of an immune response. Strong cellular and humoral immune responses against vaccinia virus are expected so that protecting immune memory could be developed without the hazard of being exposed to live virus.

该副本是利用资源的众多研究子项目之一 由NIH/NCRR资助的中心赠款提供。对该子弹的主要支持 而且，副投影的主要研究员可能是其他来源提供的 包括其他NIH来源。 列出的总费用可能 代表subproject使用的中心基础架构的估计量， NCRR赠款不直接向子弹或副本人员提供的直接资金。 在发生生物恐怖袭击的情况下，天花是一种威胁，因此，针对该病毒开发安全的疫苗很重要。有一个重要的非接种疫苗的免疫功能低下的人群无法通过当前的疫苗安全接种疫苗。在其中，是移植患者，在免疫疗法，孕妇和老年人的情况下。我们对开发一种更安全的疫苗感兴趣，该疫苗可以保护人口免受可能的攻击。我们假设佐剂介导的分子抗原免疫反应增强将控制感染小鼠的病毒负担。但是，由于天花只是可能的威胁之一，因此该项目将提供一个出色的模型来测试分子佐剂的假设，该假设将有助于建立一个可以应用于其他类别A，B和C优先级病原体的系统。小鼠将用质粒DNA进行免疫，该质粒DNA编码为谷谷病毒蛋白C7L和A55R基因，这些基因是已知会产生细胞免疫反应的非结构蛋白，以及最近发现的E6R和L3L核心蛋白。这些分子的免疫学尚未得到我们的最大知识。动物将与每种构建体接种疫苗，或与两个分子佐剂，趋化因子MIP-1-Alpha和另一种编码为树突状细胞特异性生长因子FLT-3L编码的构建体接种，这是一种已知的组合，可增加树突状细胞的募集和膨胀。据我们所知，在天花疫苗接种的背景下，这种分子佐剂的这种结合从未进行过测试。每个抗原的细胞和体液介导的免疫反应将分别由ELISPOT和ELISA确定。将进行每种抗原的分子映射以鉴定显性表位。动物将受到致命剂量的牛ac病毒的挑战，病毒载量将通过qPCR分析确定。记忆，激活和增殖将确定以检测每种抗原对诱导免疫反应的持久作用。预计对疫苗病毒的强细胞和体液免疫反应有效，因此可以开发免疫记忆力，而不会危害暴露于活病毒。