Qualification of Assays to Measure Human T-cell Responses Against Mycobacterial Lipid Antigens

测量人类 T 细胞对分枝杆菌脂质抗原反应的测定方法的资格

• 批准号: 9312745
• 负责人: CHETAN SESHADRI
• 金额: $ 36.15万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-07-07 至 2021-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9312745
• 关键词: Adult; Anti-Bacterial Agents; Antigen-Presenting Cells; Antigens; Attenuated; BCG Vaccine; Biological Assay; Biological Models; Blood; Blood donor; CD1 Antigens; CD8B1 gene; Cause of Death; Cell Line; Cell Wall; Cells; Cessation of life; Child; Clinical Trials; Collaborations; Computational Biology; Data; Development; Disease; Epidemic; Flow Cytometry; Funding; Genes; Genetic; Genus Mycobacterium; Goals; HIV; HIV Vaccine Trials Network; HIV vaccine; HLA-DR Antigens; Human; Immune; Immune response; Immune system; Immunity; Immunologics; Immunology; Interferon Type II; Interleukin-2; Interleukin-4; International; K562 Cells; Knowledge; Lipids; Major Histocompatibility Complex; Major Histocompatibility Complex Gene; Mammals; Measures; Memory; Methods; Mycobacterium bovis; Mycobacterium tuberculosis; Peptides; Phase I Clinical Trials; Phenotype; Population Study; Proteins; Pulmonary Tuberculosis; Recombinants; Reproducibility; Rest; Sampling; South Africa; South African; Specificity; System; T cell response; T-Lymphocyte; TNF gene; TNFSF5 gene; Tuberculosis; Tuberculosis Vaccines; United States; Vaccination; Vaccines; Vertebrates; Waxes; Whole Cell Vaccine; antigen binding; assay development; base; cohort; cytokine; efficacy study; experience; high dimensionality; immunogenicity; interleukin-22; mycobacterial; novel vaccines; prevent; protein complex; public health relevance; receptor; stable cell line; tool; vaccination against tuberculosis; vaccine candidate; vaccine trial; virtual

PROJECT SUMMARY (ABSTRACT)   Mycobacterium tuberculosis (M.tb) was responsible for more than 1.5 million deaths in 2013, making it a leading infectious cause of death worldwide. Currently, Mycobacterium bovis bacille Calmette-Guérin (BCG) is the only licensed vaccine for tuberculosis. BCG provides protection against disseminated forms of the disease in children but is inconsistent in preventing the development of pulmonary tuberculosis in adults. Since adults with pulmonary tuberculosis are highly infectious, control of the epidemic will not be achieved with BCG, and new vaccines are urgently needed. There are a number of vaccines under development, including recombinant BCG and attenuated M.tb strains. A successful immune response to M.tb depends critically on T- cells, which are typically activated by bacterial peptide antigens bound to major histocompatibility complex (MHC) molecules. Like BCG, candidate whole cell vaccines are poly-antigenic and contain both peptide and non-peptide antigens that are recognized by human T cells. Mycobacterial cell wall lipids have conclusively been shown to activate human T cells when bound to CD1 proteins on antigen-presenting cells. The CD1 system is conserved among mammals and mostly absent from other vertebrates, suggesting it evolved to perform an important function that is non-redundant with the MHC system. However, there are no validated assays to measure human T-cell responses against non-peptide antigens, such as lipids. We have established partnerships with experts in lipid antigen discovery, flow cytometry, vaccines, and computational biology to tackle this technical challenge over the last five years. We recently developed soluble CD1 tetramers and an activation-based T-cell profiling assay as tools to facilitate population-based studies of T-cell responses to mycobacterial lipids. We have also developed unbiased computational approaches to the analysis of high-dimensional flow cytometry data. In Aim 1, we will optimize and qualify an assay using lipid- loaded CD1 tetramers to study the memory phenotype and activation status of lipid-specific T cells at rest. In Aim 2, we will optimize and qualify a complementary activation-based assay to study the effector functions of T cells activated by lipid antigens. Unlike MHC proteins, CD1 proteins are virtually non-polymorphic so these assays can be applied independent of genetic background. In Aim 3, we will use both assays to study the effect of mycobacterial vaccination on lipid-specific T-cell responses in humans using BCG as a model system. Major collaborators on this proposal are the HIV Vaccine Trials Network (HVTN) and the South African Tuberculosis Vaccine Initiative (SATVI), which have extensive experience in developing immune-based assays for candidate HIV and tuberculosis vaccine trials, respectively. By the end of the funding period, we will have qualified a suite of assays that will supplement existing approaches to identifying correlates of protective immunity in efficacy studies of whole cell mycobacterial vaccines.

项目摘要(摘要) -- 结核分枝杆菌(M.tb)在2013年造成了150多万人死亡，使其成为 这是世界范围内主要的传染病死亡原因。目前，牛分支杆菌卡介苗(BCG) 是唯一获得许可的结核病疫苗。卡介苗提供保护，防止传播形式的 在预防成人肺结核方面并不一致，但在儿童疾病中的作用却不一致。自.以来 成人肺结核传染性强，卡介苗控制不了疫情， 而且迫切需要新的疫苗。有一些疫苗正在开发中，包括 重组卡介苗和结核分枝杆菌减毒株。对结核分枝杆菌的成功免疫反应关键取决于T- 细胞，通常由与主要组织相容性复合体结合的细菌肽抗原激活 (MHC)分子。像卡介苗一样，候选的全细胞疫苗是多抗原的，同时含有多肽和 被人类T细胞识别的非肽类抗原。分枝杆菌细胞壁脂类具有决定性的 已被证明在与抗原呈递细胞上的CD1蛋白结合时能激活人类T细胞。CD1 该系统在哺乳动物中是保守的，在其他脊椎动物中几乎不存在，这表明它进化到了 使用MHC系统执行非冗余的重要功能。然而，目前还没有经过验证的 用来测量人类T细胞对非肽抗原的反应，如脂类。我们有 与脂类抗原发现、流式细胞术、疫苗和计算领域的专家建立了合作伙伴关系 生物学在过去五年里解决了这一技术挑战。我们最近开发出了可溶性CD1 四聚体和基于激活的T细胞图谱分析作为促进基于人群的T细胞研究的工具 对分枝杆菌脂类的反应。我们还开发了无偏见的计算方法来 高维流式细胞仪数据分析。在目标1中，我们将优化和鉴定一种使用脂类- 负载CD1四聚体，研究静息状态下脂质特异性T细胞的记忆表型和激活状态。在……里面 目的2，我们将优化和鉴定一种基于互补激活的方法来研究T细胞的效应功能 被脂类抗原激活的细胞。与MHC蛋白不同，CD1蛋白实际上是非多态的，所以这些 检测可以独立于遗传背景进行应用。在目标3中，我们将使用这两种分析方法来研究 以卡介苗为模型系统研究分枝杆菌疫苗对人类脂质特异性T细胞应答的影响。 该提案的主要合作者是艾滋病毒疫苗试验网络(HVTN)和南非 结核病疫苗倡议(SATVI)，它们在开发基于免疫的检测方面拥有丰富的经验 分别用于候选艾滋病毒和结核病疫苗试验。到资助期结束时，我们将拥有 鉴定了一套分析方法，将补充现有的方法来确定保护性疾病的相关性 全细胞分枝杆菌疫苗效力研究中的免疫。