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）：疟疾根除努力的成功取决于开发安全有效的疟疾疫苗，从而促进了完全保护感染的疫苗。迄今为止，开发这种供全球使用的疫苗的努力没有成功。然而，用亚基RT，S/AS02疫苗疫苗接种编码恶性疟原虫外孢菌菌蛋白的疫苗可显着降低非洲儿童中严重疟疾，并首次证明抗抗疫苗接种疟疾疟疾亚基疫苗的发展是可行的。我们知道，活体衰减的疟原虫会产生完整和持久的CD8+细胞毒性T淋巴细胞（CTL）介导的小鼠和人类的保护，但是这种疫苗制剂并不容易在田间部署。保护对CTL靶向的抗原的保护依赖性表明，此类细胞的靶标将成为理想的亚基疫苗抗原。以前在整个生物体范围内寻找此类靶标是由于减弱的寄生虫疫苗包含成千上万个不同的免疫靶标，导致了复杂的多特异性CTL反应 - 到目前为止，没有能够在这种复杂反应中识别出离散的抗原靶标的方法。为了解决这个问题，我将采用一种高通量方法来有效地筛选出接触疟疾小鼠的CTL的数千个候选靶标，以破译赋予对疟疾免疫力的关键CTL反应。我假设保护性CTL反应包括广泛的抗原特异性，并包括在肝脏和红细胞阶段表达的寄生虫蛋白。这些靶抗原可以使用这种高通量小型驱动的筛选方法迅速鉴定。在特定目标1中，我将这种方法与小鼠的不同疫苗接种模型相结合，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。我还将使用互补的实验和计算机方法来指导集中的候选目标库的开发。在特定的目标2中，我将通过疟疾感染的红细胞评估CTL的交叉染色，并确定此类靶标如何引起跨阶段CTL免疫。这项工作中定义的免疫靶标可用于从暴露于疟疾的人类中合理地测试CTL，以定义人类中的多性保护性CTL库。拟议的方法将作为识别保护性CTL靶标的强大范式，通常可以应用于疟疾和其他感染。该申请介绍了PA-10-059（K08职业发展奖），并旨在确保出色的指导，强大的机构支持和成功的合作。 叙事全球根除疟疾需要有效的抗感染疟疾疫苗。这种疫苗的开发需要从疟原虫表达的数千种蛋白质中鉴定出离散抗原靶标的。我们在这里建议研究小鼠不同免疫方法引起的CTL，并通过使用高通量筛查技术和其他尖端方法来鉴定CTL反应来鉴定保护性抗原，以加速疟疾疫苗的发育。