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: 10265651
• 负责人: Alessandro Sette
• 金额: $ 29.28万
• 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/10265651
• 关键词: Acute; Aftercare; Aliquot; Area; Attenuated; Attenuated Vaccines; 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相关的免疫特征（IMS），即登革热病毒（DENV） 和结核分枝杆菌（MTB）。在当前周期中，我们成功实现了目标 在原始提案中的第四，并表征了与DENV和MTB相关的记忆T细胞IMS ：1）自然免疫和/或感染控制，2）活性和严重疾病，以及3） 给药许可或实验疫苗。我们的小组研究了DENV和MTB几个 由于以下原因的年份：1）它们都是当前的全球健康问题，2）记忆T细胞是 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 在体内被足够数字检测到，使其可以用于“ OMICS”研究，而无需 体外扩张，以及4）与自然感染/免疫和严重疾病相关的人类标本 很容易访问大量。 HIPC研究格式非常适合完成 拟议的任务。每个项目和核心都是并且至关重要地取决于 程序。在HIPC的背景下，我们能够实现协同作用，规模经济的利益以及 将研究人员组成不同专家。 结果，我们开发了一个临界质量，该质量被广泛利用在拟议的扩展中，基于提议的扩展 在：1）一支经验丰富的调查员团队，2）与该领域领导者的临床合作， 3）广泛的表位，这些表位受到各种不同HLA I类和II分子的限制 允许对具有前所未有的精度的记忆T细胞进行分析，4）与特定T细胞相关的IMS 代表免疫力主要参与者的子集，以及5）建立了“ OMICS”测定和分析管道 生成与DENV和MTB有关的IMS。 在此拟议的扩展中，我们计划建立在DENV和MTB中T细胞IMS的定义上 通过扩大重要性和维度，在当前的5年资金期间完成 在多个但相关和协同的方向上进行观察。如上所述，工作 在当前周期中进行的与自然感染中特定T细胞亚群相关的IMS进行的 疫苗接种和急性/严重疾病。同样很明显，这些T细胞子集本身就是 异质。在这里，我们将利用单细胞测序来“向下钻”并建立特定的IMS 负责这种异质性的细胞类型。第二个维度将通过考虑其他 细胞子集。对于DENV，这将通过确定整个未分流PBMC的IMS来实现 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 我们发现对细菌有反应的非常规CD4-CD8双（Double-double-double-double-dygatigation T细胞） 裂解物，但不是常规的肽表位。