Human immune signatures of Dengue virus and Mycobacterium Tuberculosis exposure in infection, disease and vaccination

感染、疾病和疫苗接种中登革热病毒和结核分枝杆菌暴露的人体免疫特征

• 批准号: 10056696
• 负责人: Alessandro Sette
• 金额: $ 272.15万
• 依托单位: LA JOLLA INSTITUTE FOR IMMUNOLOGY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-06-01 至 2022-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10056696
• 关键词: Acute; Aftercare; Aliquot; Area; Attenuated; Attenuated Live Virus Vaccine; Bacille Calmette-Guerin vaccination; Biological Assay; CD8B1 gene; Cells; Characteristics; Clinical; Collaborations; Collection; Containment; Convalescence; Dengue Infection; Dengue Vaccine; Dengue Virus; Development; Diagnosis; Dimensions; Disease; Elements; Epitopes; Funding; Generations; Goals; Heterogeneity; Human; Immune; Immune response; Immunity; Immunologic Memory; In Vitro; Individual; Infection; Infection Control; Memory; Mission; Mycobacterium tuberculosis; Natural Immunity; Pathology; Peptides; Peripheral Blood Mononuclear Cell; Population; Predisposition; Research Personnel; Resolution; Rest; Sampling; Specimen; T cell response; T memory cell; T-Lymphocyte; T-Lymphocyte Epitopes; T-Lymphocyte Subsets; Tuberculosis; Tuberculosis Vaccines; Vaccination; Vaccines; Work; analysis pipeline; antigen-specific T cells; bacterial lysate; base; cell type; data management; experience; global health; human disease; latent infection; memory CD4 T lymphocyte; novel; pathogen; programs; response; single cell analysis; single cell sequencing; single-cell RNA sequencing; synergism; transcriptomics; γδ T cells

The fundamental premise of our application is to provide unbiased, non-hypothesis driven analyses of immune signatures (IMS) associated with two important human diseases, namely dengue virus (DENV) and Mycobacterium tuberculosis (Mtb). In the current cycle, we successfully accomplished the goals set forth in the original proposal and have characterized memory T cell IMS associated with DENV and Mtb in the context of: 1) natural immunity and/or control of infection, 2) active and severe disease, and 3) administration of licensed or experimental vaccines. Our group has studied DENV and Mtb for several years for the following reasons: 1) they are both current global health problems, 2) memory T cells are fundamentally associated with protective immunity for these diseases, 3) antigen-specific T cell responses are detected in sufficient numbers ex vivo, making them accessible for “omics” studies without the need for in vitro expansion, and 4) human specimens associated with natural infection/immunity and severe disease are easily accessible in large numbers. The HIPC study format was ideally suited to accomplish the proposed mission. Each project and core was and remains critically dependent on other elements of the program. In the context of HIPC, we were able to realize synergies, benefit from economies of scale, and bring groups of investigators with different expertise together. As a result, we developed a critical mass that is extensively leveraged in the proposed extension, based on: 1) an established team of experienced investigators, 2) clinical collaborations with leaders in the field, 3) an extensive collection of epitopes restricted by a variety of different HLA class I and II molecules to allow an analysis of memory T cells with unprecedented precision, 4) IMS associated with specific T cell subsets representing key players in immunity, and 5) established “omics” assays and analysis pipelines to generate IMS related to DENV and Mtb. In this proposed extension, we plan to build on the definition of the T cell IMS in DENV and Mtb accomplished in the current 5-year funding period by expanding the significance and the dimensionality of the observations in multiple, yet related and synergistic, directions. As mentioned above, the work performed in the current cycle identified IMS associated with specific T cell subsets in natural infection, vaccination, and acute/severe disease. It is also clear that these T cell subsets are themselves heterogeneous. Here, we will utilize single-cell sequencing to “drill down” and establish IMS of the specific cell types responsible for this heterogeneity. The second dimension will “drill up” by considering additional cell subsets. For DENV, this will be accomplished by determining the IMS of whole unfractionated PBMC stimulated by epitopes recognized by DENV memory T cells. For Mtb, we will extend our focus to unconventional CD4-CD8-double-negative T cells which we have found to be responsive to bacterial lysates but not conventional peptide epitopes.

我们应用程序的基本前提是提供公正的、非假设驱动的分析 与两种重要人类疾病（即登革热病毒（DENV））相关的免疫特征（IMS） 和结核分枝杆菌（Mtb）。本周期，我们圆满完成既定目标 在最初的提案中提出，并在中描述了与 DENV 和 Mtb 相关的记忆 T 细胞 IMS 背景：1) 自然免疫力和/或感染控制，2) 活动性和严重疾病，以及 3) 许可或实验疫苗的管理。我们小组对 DENV 和 Mtb 进行了多次研究 原因如下：1）它们都是当前的全球健康问题，2）记忆 T 细胞 从根本上与这些疾病的保护性免疫相关，3) 抗原特异性 T 细胞反应 离体检测到足够数量，使它们无需进行“组学”研究即可进行 体外扩增，4) 与自然感染/免疫和严重疾病相关的人体标本 很容易大量获得。重债穷国研究形式非常适合完成 拟议的任务。每个项目和核心过去和现在都严重依赖于项目的其他要素 程序。在重债穷国的背景下，我们能够实现协同效应，从规模经济中受益， 将具有不同专业知识的研究人员小组聚集在一起。 因此，我们开发了一个临界质量，在拟议的扩展中得到广泛利用，基于 1) 经验丰富的研究人员组成的成熟团队，2) 与该领域领导者的临床合作， 3) 受多种不同 HLA I 类和 II 类分子限制的表位的广泛集合 允许以前所未有的精度分析记忆 T 细胞，4) 与特定 T 细胞相关的 IMS 代表免疫关键参与者的子集，以及 5) 建立“组学”测定和分析管道 生成与 DENV 和 Mtb 相关的 IMS。 在这个拟议的扩展中，我们计划以 DENV 和 Mtb 中 T 细胞 IMS 的定义为基础 通过扩大以下项目的重要性和维度，在当前的 5 年资助期内完成了这项工作： 多个但相关且协同的方向的观察。如上所述，工作 在当前周期中进行的识别 IMS 与自然感染中的特定 T 细胞亚群相关， 疫苗接种和急性/严重疾病。同样清楚的是，这些 T 细胞亚群本身就是 异质。在这里，我们将利用单细胞测序来“深入”并建立特定的IMS 造成这种异质性的细胞类型。第二个维度将通过考虑额外的内容来“钻取” 细胞亚群。对于 DENV，这将通过确定整个未分割 PBMC 的 IMS 来完成 由 DENV 记忆 T 细胞识别的表位刺激。对于 Mtb，我们将把重点扩展到 我们发现非常规 CD4-CD8 双阴性 T 细胞对细菌有反应 裂解物，但不是常规肽表位。