Identification of responding CD8+ T cells and novel protective epitopes following

鉴定响应 CD8 T 细胞和新的保护性表位

• 批准号: 8523775
• 负责人: Sean C Murphy
• 金额: $ 12.69万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-22 至 2016-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8523775
• 关键词: Address; Africa; Animals; Antigen Targeting; Antigenic Specificity; Antigens; Attenuated; Biomedical Research; Blood; CD8B1 gene; Calculi; Cells; Cessation of life; Child; Collaborations; Complex; Computer Simulation; Cross-Priming; Cytotoxic T-Lymphocytes; Dependence; Development; Ensure; Epitopes; Erythrocytes; Foundations; Fred Hutchinson Cancer Research Center; Future; Goals; Human; Hybridomas; Immune Targeting; Immunity; Immunization; Immunology; Infection; Investigation; K-Series Research Career Programs; Kinetics; Libraries; Life; Liver; Major Histocompatibility Complex; Malaria; Malaria Vaccines; Mass Spectrum Analysis; Mediating; Mentors; Mentorship; Methodology; Methods; Modeling; Molecular Biology; Monitor; Mus; Needles; Parasites; Peptides; Plasmodium; Plasmodium falciparum; Positioning Attribute; Postdoctoral Fellow; Preparation; Problem Solving; Proteins; Research; Research Institute; Resources; Role; Specificity; Sporozoite vaccine; Sporozoites; Staging; Subunit Vaccines; T cell response; T-Lymphocyte; Technology; Testing; Time; Universities; Vaccinated; Vaccination; Vaccine Antigen; Vaccines; Washington; Whole Organism; Work; base; career; circumsporozoite protein; design; experience; high throughput screening; novel; programs; response; screening; skills; success; tool; vaccine development; vaccinology

DESCRIPTION (provided by applicant): The success of malaria eradication efforts hinges on the development of a safe and efficacious malaria vaccine that induces complete protection against infection. To date, efforts to develop such a vaccine for worldwide use have been unsuccessful. However, vaccination with the subunit RTS,S/AS02 vaccine encoding the Plasmodium falciparum circumsporozoite protein significantly reduced deaths due to severe malaria in Africa children and demonstrated for the first time that development of an anti-infection malaria subunit vaccine is feasible. We know that live-attenuated Plasmodium parasites yield complete and long-lasting CD8+ cytotoxic T lymphocyte (CTL)-mediated protection in mice and humans, but such vaccine preparations are not easily deployed in the field. The dependence of protection on antigens targeted by CTLs indicates that the targets of such cells would make ideal subunit vaccine antigens. The search for such targets was previously impossible on a whole organism scale because attenuated parasite vaccines contain thousands of different immune targets leading to complex polyspecific CTL responses - until now there was no methodology capable of identifying discrete antigenic targets in the midst of such a complex response. To solve this problem, I will employ a high-throughput method to efficiently screen thousands of candidate targets against CTLs from malaria-exposed mice in order to decipher the key CTL responses that confer immunity against malaria. I hypothesize that protective CTL responses encompass a wide range of antigenic specificities and include parasite proteins expressed in both liver and erythrocyte stages. These target antigens can be rapidly identified using this high-throughput minigene-driven screening approach. In Specific Aim 1, I will combine this approach with different vaccination models in mice to assess the kinetics and diversity of CTLs and identify cognate target antigens associated with protection. I will also use complementary experimental and in silico approaches to guide development of a focused candidate target library. In Specific Aim 2, I will evaluate cross-priming of CTLs by malaria-infected erythrocytes and determine how such targets elicit cross-stage CTL immunity. Immune targets defined in this work can later be used to rationally test CTLs from malaria-exposed humans to define a polyspecific protective CTL repertoire in humans. The proposed approach will serve as a powerful paradigm for identifying protective CTL targets and can be generally applied to malaria and other infections. This application addresses PA-10-059 (K08 Career Development Award) and was crafted to ensure excellent mentorship, strong institutional support and successful collaborations. NARRATIVE Global eradication of malaria requires an efficacious anti-infection malaria vaccine. Development of such a vaccine requires the identification of discrete antigenic targets from among thousands of proteins expressed by malaria parasites. We propose here to study CTLs induced by different immunization approaches in mice and identify protective antigens by profiling CTL responses using high-throughput screening technologies and other cutting-edge approaches to accelerate malaria vaccine development.

描述（由申请人提供）：根除疟疾工作的成功取决于开发出一种安全有效的疟疾疫苗，该疫苗可提供全面的感染保护。迄今为止，开发这种供全球使用的疫苗的努力尚未成功。然而，接种编码恶性疟原虫环子孢子蛋白的亚单位RTS，S/AS02疫苗显着减少了非洲儿童因严重疟疾导致的死亡，并首次证明开发抗感染疟疾亚单位疫苗是可行的。我们知道，减毒活疟原虫寄生虫可以在小鼠和人类中产生完整且持久的 CD8+ 细胞毒性 T 淋巴细胞 (CTL) 介导的保护，但此类疫苗制剂并不容易在现场部署。保护作用对 CTL 靶向抗原的依赖性表明，此类细胞的靶标将成为理想的亚单位疫苗抗原。以前在整个生物体范围内寻找此类靶标是不可能的，因为减毒寄生虫疫苗包含数千个不同的免疫靶标，导致复杂的多特异性 CTL 反应 - 直到现在，还没有能够在如此复杂的反应中识别离散抗原靶标的方法。为了解决这个问题，我将采用高通量方法，有效筛选来自疟疾暴露小鼠的数千个针对 CTL 的候选靶点，以破译赋予抗疟疾免疫力的关键 CTL 反应。我假设保护性 CTL 反应涵盖广泛的抗原特异性，包括在肝脏和红细胞阶段表达的寄生虫蛋白。使用这种高通量小基因驱动的筛选方法可以快速鉴定这些靶抗原。在具体目标 1 中，我将将此方法与小鼠的不同疫苗接种模型结合起来，以评估 CTL 的动力学和多样性，并识别与保护相关的同源靶抗原。我还将使用补充实验和计算机方法来指导重点候选目标库的开发。在具体目标 2 中，我将评估感染疟疾的红细胞对 CTL 的交叉启动，并确定这些目标如何引发跨阶段 CTL 免疫。这项工作中定义的免疫目标随后可用于合理测试来自疟疾暴露人群的 CTL，以定义人类的多特异性保护性 CTL 库。所提出的方法将作为识别保护性 CTL 靶点的强大范例，并且可普遍应用于疟疾和其他感染。该申请针对 PA-10-059（K08 职业发展奖），旨在确保出色的指导、强大的机构支持和成功的合作。 叙述 全球消灭疟疾需要有效的抗感染疟疾疫苗。开发这种疫苗需要从疟原虫表达的数千种蛋白质中鉴定出离散的抗原靶点。我们在此建议研究小鼠中不同免疫方法诱导的 CTL，并通过使用高通量筛选技术和其他尖端方法分析 CTL 反应来识别保护性抗原，以加速疟疾疫苗的开发。