Development of novel subbunit vaccine targeting mutiple alphaviruses

开发针对多种甲病毒的新型亚单位疫苗

• 批准号: 8465797
• 负责人: KENNETH E OLSON
• 金额: $ 38.04万
• 依托单位: COLORADO STATE UNIVERSITY
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至 2015-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8465797
• 关键词: Adjuvant; Adsorption; Aerosols; Alphavirus; Antibodies; Antibody Formation; Antigens; B-Lymphocytes; Baculoviruses; Categories; Cells; Collaborations; Complex; Culicidae; DNA; Development; Disease; Disease Outbreaks; Eastern Equine Encephalitis Virus; Encephalitis; Exposure to; Glycoproteins; Human; Immune response; Immunity; Immunization; Immunoglobulin A; Immunoglobulin G; Immunoglobulin M; Insecta; Liposomes; Longevity; Maps; Mus; National Institute of Allergy and Infectious Disease; Natural Immunity; Pathogenesis; Pathway interactions; Penetration; Polysaccharides; Proteins; Recombinants; Research; Research Project Grants; Resources; Ross river virus; Route; Sindbis Virus; Subunit Vaccines; System; T-Lymphocyte; T-Lymphocyte Epitopes; Technology; Testing; Togaviridae; Vaccine Adjuvant; Vaccines; Viral; Virus; Western Equine Encephalitis Virus; Widespread Disease; base; biodefense; chikungunya; glycosylation; immunoregulation; mouse model; neutralizing antibody; novel; novel vaccines; response; subcutaneous; virus envelope

Eastern and Western equine encephalitis viruses (EEEV and WEEV; Alphavirus; Togaviridae) are mosquitoborne alphaviruses causing severe encephalitis in humans. There are no human vaccines for alphaviruses in case of widespread disease. We have previously developed mouse models describing pathogenesis of these alphaviruses and have shown that cationic-liposome-DNA complexes (CLDC) elicit protective activation of innate immunity in mice following WEEV challenge. In this proposal, CLDC-based vaccine platforms (LANAC) will be developed that include E2-E1 and E1 glycoproteins having novel N-glycans. We will test the hypothesis that immunization with WEEV E2-E1 or E1-based LANAC vaccines elicits protective immunity against multiple alphaviruses. The specific aims of the proposal are as follows: Aim 1) Construct and produce recombinant forms of the WEEV envelope glycoproteins, E1 and E2-E1 using a conventional baculovirus-insect cell system and a baculovirus-insect cell system with a humanized protein A/-glycosylation pathway. Aim 2) Evaluate protection provided by recombinant envelope WEEV glycoproteins (invertebratetype A/-glycans or vertebrate-type A/-glycans) as subunit vaccines in mice when used in combination with CLDC's. Protection will be evaluated following subcutaneous and mucosal immunization in the mouse model and aerosol-, subcutaneous-, and mosquito-delivered virus challenge. Aim 3) Identify the humoral and cellular immunological mechanisms responsible for the most efficient protection of mice following exposure to the WEEV E1 and E2-E1 glycoprotein vaccines with CLDC adjuvant. These immunological responses include quantifying IgG, IgM and IgA antibody and neutralizing antibody titers and mapping potentially protective T cell epitopes to WEEV glycoproteins. We will also compare immune responses to E1 and E2-E1 glycoproteins in immunized CD1, B6, mice which lack B-cells, and T-cell depleted mice. Aim 4) Characterize the ability of WEEV E1 glycoprotein to provide cross-protection in mice to other alphaviruses. Cross protective glycoproteins with CLDC's will be evaluated for EEEV, VEEV, and Sindbis virus (SINV) at CSU and Chikungunya and Ross River viruses (CHIKV and RRV) at UNC-Chapel Hill as part of a collaboration with Dr. Robert E. Johnston's lab and SERCEB. We will evaluate protection against multiple virus challenge routes and evaluate the longevity of immunological responses and protection. This research project fits within the RMRCE Integrated Research Focus on Immunomodulation, Adjuvants and Vaccines, and will interact with RP1.5, RP1.6, RP 3.3 and utilize the resources of Core C and E.

东马和西马脑炎病毒（EEEV和WEEV；α型甲虫； togaviridae）是蚊子 α病毒引起人类严重的脑炎。没有针对α病毒的人类疫苗 广泛疾病的病例。我们以前已经开发了描述的小鼠模型 这些α病毒，并表明阳离子 - 脂肪组-DNA复合物（CLDC）引起保护性 在WEEV挑战之后，小鼠先天免疫的激活。在此提案中，基于CLDC的疫苗 将开发平台（LANAC），其中包括具有新型N-聚糖的E2-E1和E1糖蛋白。我们 将测试以WEEV E2-E1或基于E1的LANAC疫苗免疫的假设 对多种α病毒的免疫力。该提案的具体目的如下：目标1）构建 并使用常规的WEEV包膜糖蛋白E1和E2-E1产生重组形式 杆状病毒诱导细胞系统和带人源化蛋白A/ - 糖基化的杆状病毒 - 诱导细胞系统 路径。目标2）评估重组信封WEEV糖蛋白提供的保护 与小鼠中的亚糖或脊椎动物型A/-Glycans一起作为亚基疫苗，当 CLDC的。在小鼠模型中皮下和粘膜免疫后，将评估保护 和气溶胶，皮下和蚊子传递的病毒挑战。目标3）识别体液和 暴露后，负责最有效保护小鼠的细胞免疫机制 使用CLDC辅助剂到WEEV E1和E2-E1糖蛋白疫苗。这些免疫学反应 包括量化IgG，IgM和IgA抗体以及中和抗体滴度以及可能映射 保护性T细胞表现为WEEV糖蛋白。我们还将比较与E1和E2-E1的免疫反应 缺乏B细胞和T细胞耗尽的小鼠的免疫CD1，B6，B6的糖蛋白。目标4）特征 WEEV E1糖蛋白在小鼠向其他α病毒中提供交叉保护的能力。叉 将对CSU的EEEV，VEEV和SINDBIS病毒（SINV）评估具有CLDC的保护性糖蛋白 以及UNC-Chapel Hill的Chikungunya和Ross River Viruse（Chikv和RRV）作为合作的一部分 与Robert E. Johnston博士的实验室和Serceb一起。我们将评估防止多种病毒挑战的保护 路线和评估免疫反应和保护的寿命。该研究项目适合 在RMRCE综合研究中，重点是免疫调节，佐剂和疫苗，并将 与RP1.5，RP1.6，RP 3.3相互作用，并利用Core C和E的资源。