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: 8261422
• 负责人: KENNETH E OLSON
• 金额: $ 37.17万
• 依托单位: COLORADO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-05-01 至 2014-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8261422
• 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.

东西方马脑炎病毒（EEEV 和 WEEV；甲病毒；披膜病毒科）是蚊媒传播的 甲病毒引起人类严重脑炎。目前还没有针对甲病毒的人类疫苗 疾病广泛传播的情况。我们之前开发了描述发病机制的小鼠模型 这些甲病毒并已表明阳离子-脂质体-DNA 复合物 (CLDC) 会引发保护性 WEEV 攻击后小鼠先天免疫的激活。在该提案中，基于 CLDC 的疫苗 将开发包括具有新型 N-聚糖的 E2-E1 和 E1 糖蛋白的平台 (LANAC)。我们 将检验以下假设：使用 WEEV E2-E1 或基于 E1 的 LANAC 疫苗进行免疫会产生保护性 对多种甲病毒的免疫力。该提案的具体目标如下： 目标 1) 构建 并使用常规方法生产 WEEV 包膜糖蛋白 E1 和 E2-E1 的重组形式 杆状病毒-昆虫细胞系统和具有人源化蛋白A/-糖基化的杆状病毒-昆虫细胞系统 途径。目标 2) 评估重组包膜 WEEV 糖蛋白（无脊椎动物型 A/-聚糖或脊椎动物型 A/-聚糖）作为小鼠亚单位疫苗与 CLDC 的。将在小鼠模型中进行皮下和粘膜免疫后评估保护作用 以及气溶胶、皮下和蚊子传播的病毒挑战。目标 3) 识别体液和 细胞免疫机制负责在暴露后最有效地保护小鼠 含有 CLDC 佐剂的 WEEV E1 和 E2-E1 糖蛋白疫苗。这些免疫反应 包括量化 IgG、IgM 和 IgA 抗体以及中和抗体滴度和潜在绘图 WEEV 糖蛋白的保护性 T 细胞表位。我们还将比较对 E1 和 E2-E1 的免疫反应 免疫 CD1、B6、缺乏 B 细胞的小鼠和 T 细胞耗尽的小鼠中的糖蛋白。目标 4) 表征 WEEV E1 糖蛋白为小鼠提供针对其他甲病毒的交叉保护的能力。叉 带有 CLDC 的保护性糖蛋白将在科罗拉多州立大学针对 EEEV、VEEV 和 Sindbis 病毒 (SINV) 进行评估 作为合作的一部分，北卡罗来纳大学教堂山分校的基孔肯雅病毒和罗斯河病毒（CHIKV 和 RRV） 与 Robert E. Johnston 博士的实验室和 SERCEB 合作。我们将评估针对多种病毒挑战的防护能力 路线并评估免疫反应和保护的寿命。该研究项目适合 RMRCE 综合研究重点是免疫调节、佐剂和疫苗，并将 与RP1.5、RP1.6、RP 3.3交互并利用Core C和E的资源。