Collective Responses to Malaria Vaccination

摘要 项目的总体目标是确定接种减毒活疫苗后的免疫反应的关键特征。 恶性疟原虫子孢子(APfSPZ)，可产生不育性免疫。我们假设这是 免疫是由于细胞和体液对Pf抗原(Ags)的反应，这种反应在 感染的肝期(LS)。我们对先天免疫和获得性免疫进行了详细的纵向分析。 使用尖端免疫学分析、相关信息学和系统免疫学进行回应。我们 将确定SPZ疫苗诱导的与疟疾保护相关的细胞和体液反应 幼稚和疟疾经历过的对象。互补的正交分析工具和方法将 确定相关的免疫细胞亚群、抗原受体(AR)谱系和保护性的特异性 AGS.免疫细胞和免疫产物的功能特征，包括抗体，与 将确定和验证保护措施。这将通过以下方式实现：目标1：确定一种特殊疫区疫苗 诱导与保护相关的细胞和体液反应。无偏和抗原特异性的细胞 描述将描述与疫苗接种和CHMI相关的保护反应的动力学。后续行动 分析将描述通过质量细胞术(CyTOF)识别的Ag特异性适应性和先天细胞反应， 多重pMHC/Ag多聚体染色和多组单细胞测序确定保护- 相关的基因和蛋白质表达特征和功能性抗体反应，这将被扩展 在以后的目标中。目的2：确定保护相关的T和B细胞抗原受体(AR)在关键时间段 接种疫苗和CHMI。来自T和B细胞的天然配对和单个ARs链将被测序至 确定谱系属性并鉴定疫苗诱导的Pf抗原特异性受体。细胞转录图谱 而Ag的特异性将通过联合单细胞和TCR/BCR序列分析联系起来。新型AR-Ag和 抗体的特异性将通过酵母展示方法进行鉴定。该目标将识别T和B细胞ARs和 与保护相关的抗体抗原特异性。目标3：确定的功能特征 提供保护的细胞和抗体反应。分选的反应性T细胞将通过抗原特异性检测 细胞内细胞因子染色(ICS)、激活诱导标记(AIM)和单细胞序列 基于多元组学分析。单抗的生物物理和功能分析将用于 评估B细胞在保护中的作用。综合项目数据将进行综合多因素分析 利用已建立的平台进行生物信息分析、管理和共享。总体而言，该项目将确定关键 ASPZ PF疫苗复合抗原成分的复杂免疫反应特征 以及与保护相关的效应器过程，并提供一套与保护相关的免疫相关 对疟疾的免疫力，以指导疫苗开发。

SUMMARY The overall project goal is to identify key characteristics of immune responses to vaccination with attenuated Plasmodium falciparum sporozoites (aPfSPZs) which results in sterilizing immunity. We hypothesize that this immunity is due to cellular and humoral responses to specific to Pf antigens (Ags) that are presented during the liver stage (LS) of infection. We propose a detailed longitudinal analysis of innate and adaptive immune responses using cutting-edge immunological analyses, associated informatics, and systems immunology. We will identify aSPZ vaccine-induced cellular and humoral responses that correlate with protection in malaria naïve and malaria experienced subjects. Complementary orthogonal analytical tools and approaches will identify relevant immune cell subsets, Ag receptor (AR) repertoires, lineages, and specificities to protective Ags. The functional characteristics of the immune cells and products, including antibodies, that correlate with protection will be identified and validated. This will be accomplished by: Aim 1: Identifying aSPZ vaccine induced cellular and humoral responses that are associated with protection. Unbiased and Ag specific cellular profiling will characterize the kinetics of protection associated responses to vaccination and CHMI. Follow-on analyses will profile identified Ag-specific adaptive and innate cellular responses by mass cytometry (CyTOF), multiplexed pMHC/Ag multimer staining, and multi-omic single cell sequencing to determine protection- associated gene and protein expression signatures and functional antibody responses, which will be extended in later aims. Aim 2: Identify protection associated T and B cell Ag receptor (AR) repertoires at key times after vaccinations and CHMI. Natively paired and single ARs chains from T and B cells will be sequenced to determine repertoire attributes and identify vaccine induced Pf Ag specific ARs. Cellular transcriptomic profiles and Ag specificities will be linked by joint single cell and TCR/BCR sequence analyses. Novel AR-Ag and antibody specificities will be identified by yeast display methods. This aim will identify T and B cell ARs and antibody Ag specificities that are correlated with protection. Aim 3: Identifying functional characteristics of cellular and antibody responses that confer protection. Sorted reactive T cells will be assayed by Ag-specific CyTOF-based intracellular cytokine staining (ICS), activation-induce marker (AIM), and single-cell sequence based multi-omics analyses. The biophysical and functional analyses of monoclonal antibodies will be used to assess B cell roles in protection. The comprehensive project data will undergo integrated multi-factorial bioinformatic analysis, curation and sharing using established platforms. Overall, this project will identify key characteristics of the intricate immune responses to the complex antigenic composition of aSPZ Pf vaccination and the effector processes that correlate with protection and provide a set of immune correlates of protective immunity against malaria to guide vaccine development.