Identification of targets of CD4 and CD8 T cell reactivity conserved across Sarbecoviruses and recognized across different animal species

鉴定在 Sarbecoviruses 中保守并在不同动物物种中识别的 CD4 和 CD8 T 细胞反应性靶标

• 批准号: 10420515
• 负责人: Alba Grifoni
• 金额: $ 194.61万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-02 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10420515
• 关键词: 2019-nCoV; Animals; Antigens; Architecture; Bioinformatics; Biological Assay; CD8-Positive T-Lymphocytes; COVID-19; Catalogs; Cell Line; Cellular Immunity; Complement; Coronavirus; Data; Data Set; Development; Distant; Epitopes; Evaluation; Future; Goals; Hamsters; Human; Human Cell Line; Humoral Immunities; Immunity; Immunization; Immunodominant Antigens; Infection; Laboratories; Lead; Merbecovirus; Mesocricetus auratus; Meta-Analysis; Mouse Strains; Mus; Nonstructural Protein; Open Reading Frames; Outcome; Pattern; Peptides; Phylogenetic Analysis; Polyproteins; Preclinical Testing; Protein Region; Proteins; Proteome; Rodent Model; Sarbecovirus; Sequence Homologs; T cell response; T memory cell; T-Lymphocyte; T-Lymphocyte Epitopes; Taxonomy; Testing; Update; Vaccination; Vaccine Antigen; Vaccine Design; Vaccines; Validation; Viral; Virus; Work; base; betacoronavirus; cross immunity; cross reactivity; experimental group; human subject; immunogenicity; interest; manufacturability; novel coronavirus; pandemic coronavirus; pandemic disease; predictive tools; programs; prototype; universal coronavirus vaccine; vaccine platform

PROJECT SUMMARY The overarching goal of Project 2 is to discover and design vaccine antigen candidates that confer broad cross- protective T cell immunity to multiple coronavirus (CoV) species with pandemic potential. Cross-reactivity at the T cell level has been unequivocally demonstrated between SARS-CoV-2, other Sarbecoviruses, and other more distantly related CoV in our previous studies. By incorporating strategies that explicitly elicit cellular immunity into vaccines generated by Project 3, cross-protection against future pandemics emerging from CoV reservoirs may be possible. Specific Aim 1 will define targets of CD4+ and CD8+ T cell reactivity that are broadly conserved across different Sarbecoviruses, Merbecoviruses, and other betacoronaviruses of concern by (i) identifying conserved regions across the entire CoV proteome, including all mature non-structural proteins (nsp) and open reading frames (ORFs) for each of these taxonomic groups, (ii) performing a meta-analysis of immunogenicity data to catalog known T cell epitopes for these antigens to define which antigen subregions are recognized following natural infection by different CoV species, and (iii) integrating these data to select a set of candidate conserved T cell epitope regions from each of the three taxonomic groups for experimental evaluation. This bioinformatic analysis will assign specific quantitative scores, including sequence conservation scores, human immunogenicity scores, and cross-reactivity for human T cells scores to various proteins subregions. Specific Aim 2 will test human T cells directed against epitopes from Aim 1 for their capacity to recognize homologous sequences from different Sarbecoviruses, Merbecoviruses, and other betacoronaviruses of concern by (i) synthesizing the conserved epitope regions defined in Aim 1 from prototype viruses and homologous peptides from each of the different taxonomic groups evaluated in Aim 1, (ii) eliciting T cell lines from COVID-19 convalescent human donors and human subjects with NL63 and OC43-reactive memory T cells using SARS-CoV-2-, NL63-, and OC43-derived epitopes, (iii) testing these epitope-specific T cell lines for cross- reactivity with corresponding homologous peptides from each of the three taxonomic groups using an approach developed in our laboratory to evaluate CoV cross-reactivity, and (iv) performing bioinformatic analyses to predict immunogenicity in our rodent models. Based on this experimental work, we will prioritize potential candidates, and deliver lists of candidates to the overall Program lead in Core A for incorporation into two vaccine platforms in Project 3 and testing for immunogenicity and cross-protection in Core B. This project will produce the first comprehensive evaluation of T cell immunogenicity of the entire CoV proteome across a broad cross- section of CoVs with pandemic potential using a tight integration of bioinformatics prediction and experimental validation to create antigens for vaccine design that elicit broad cross-protective cellular immunity to complement the humoral immunity elicited through Project 1.

项目概要 项目 2 的总体目标是发现和设计候选疫苗抗原，从而赋予广泛的交叉作用。 对多种具有大流行潜力的冠状病毒 (CoV) 物种的保护性 T 细胞免疫。交叉反应性 SARS-CoV-2、其他 Sarbeco 病毒和其他更多病毒之间的 T 细胞水平已得到明确证明 在我们之前的研究中，有远亲关系的冠状病毒。通过结合明确引发细胞免疫的策略 项目 3 生产的疫苗可交叉保护免受未来冠状病毒储存库中出现的流行病的影响 也许有可能。具体目标 1 将定义广泛的 CD4+ 和 CD8+ T 细胞反应性目标 在不同的 Sarbecoviruses、Merbecoviruses 和其他受关注的 β 冠状病毒中保守 (i) 识别整个 CoV 蛋白质组的保守区域，包括所有成熟的非结构蛋白 （nsp）和开放阅读框（ORF）对于每个分类群，（ii）进行荟萃分析 免疫原性数据对这些抗原的已知 T 细胞表位进行分类，从而定义哪些抗原亚区 识别出不同 CoV 物种自然感染后的结果，以及 (iii) 整合这些数据以选择一组 三个分类群中每一个的候选保守 T 细胞表位区域用于实验评估。 这种生物信息学分析将分配特定的定量分数，包括序列保守分数， 人类免疫原性评分以及人类 T 细胞与各种蛋白质亚区域的交叉反应性评分。 具体目标 2 将测试人类 T 细胞针对目标 1 中的表位的识别能力 来自不同 Sarbecoviruses、Merbecoviruses 和其他 β 冠状病毒的同源序列 值得关注的是 (i) 从原型病毒合成目标 1 中定义的保守表位区域，并且 来自目标 1 中评估的每个不同分类群的同源肽，(ii) 引发 T 细胞系 来自 COVID-19 康复期人类捐赠者和具有 NL63 和 OC43 反应性记忆 T 细胞的人类受试者 使用 SARS-CoV-2、NL63 和 OC43 衍生表位，(iii) 测试这些表位特异性 T 细胞系的交叉 使用某种方法与来自三个分类群中的每一个的相应同源肽的反应性 我们实验室开发的用于评估 CoV 交叉反应性的产品，以及 (iv) 进行生物信息分析来预测 我们的啮齿动物模型中的免疫原性。根据这项实验工作，我们将优先考虑潜在的候选人， 并将候选者名单提交给核心 A 中的总体计划负责人，以便纳入两个疫苗平台 项目 3 中的免疫原性和交叉保护测试在核心 B 中进行。该项目将产生第一个 对整个 CoV 蛋白质组的 T 细胞免疫原性进行广泛的交叉综合评估 使用生物信息学预测和预测的紧密结合来研究具有大流行潜力的 CoV 部分 实验验证为疫苗设计创建抗原，引发广泛的交叉保护性细胞 免疫来补充通过项目 1 引起的体液免疫。