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: 9152421
• 负责人: CHETAN SESHADRI
• 金额: $ 101.83万
• 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/9152421
• 关键词: Adult; Anti-Bacterial Agents; Antigen-Presenting Cells; Antigens; Attenuated; Binding; Biological Assay; Biological Models; Blood; Blood donor; CD1 Antigens; CD8B1 gene; Calmette-Guerin Bacillus; 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; Licensing; Lipids; Major Histocompatibility Complex; Major Histocompatibility Complex Gene; Mammals; Measures; Memory; Methods; Mycobacterium bovis; Mycobacterium tuberculosis; Mycobacterium tuberculosis antigens; Peptides; Phase I Clinical Trials; Phenotype; Proteins; Pulmonary Tuberculosis; Qualifying; 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; Work; antigen binding; assay development; base; cohort; cytokine; experience; immunogenicity; interleukin-22; mycobacterial; novel vaccines; population based; prevent; public health relevance; receptor; tool; vaccination against tuberculosis; vaccine candidate; vaccine development; vaccine trial

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年造成超过150万人死亡，这使得 全世界领先的感染性死亡原因。目前，牛分枝杆菌巴奇氏菌Calmette-guérin（BCG） 是唯一用于结核病的许可疫苗。 BCG提供了针对传播形式的保护 儿童疾病，但与防止成人肺结核的发展不一致。自从 患有肺结核的成年人具有高度感染性，对BCG的控制将无法实现。 迫切需要新的疫苗。正在开发许多疫苗，包括 重组BCG并减弱M.TB菌株。对M.TB的成功免疫反应取决于T- 细胞，通常由与主要组织相容性复合物结合的细菌肽抗原激活 （MHC）分子。与BCG一样 由人T细胞识别的非肽抗原。分枝杆菌细胞壁脂质具有最终的 当我们在抗原呈递细胞上与CD1蛋白结合时，我们被证明会激活人T细胞。 CD1 系统是在哺乳动物之间配置的，大多数是其他脊椎动物中不存在的，这表明它已演变为 在MHC系统中执行非冗余的重要功能。但是，没有验证 测量针对非肽抗原的人类T细胞反应的测定，例如脂质。我们有 与脂质抗原发现，流式细胞仪，疫苗和计算方面的专家建立了伙伴关系 在过去五年中应对这一技术挑战的生物学。我们最近开发了固体CD1 四聚体和基于激活的T细胞分析测定法作为促进基于人群的T细胞研究的工具 对分枝杆菌脂质的反应。我们还开发了公正的计算方法 高维流式细胞仪数据的分析。在AIM 1中，我们将使用脂质优化和合格测定法 加载的CD1四聚体研究静止时脂质特异性T细胞的记忆表型和激活状态。在 AIM 2，我们将优化并符合完整的基于激活的评估，以研究t的效应子功能 通过脂质抗原激活的细胞。与MHC蛋白不同，CD1蛋白实际上是非晶状体的，所以这些 可以独立于遗传背景来应用测定。在AIM 3中，我们将使用两种测定法来研究 分枝杆菌疫苗接种对使用BCG作为模型系统的人类脂质特异性T细胞反应的影响。 该建议的主要合作者是HIV疫苗试验网络（HVTN）和南非 结核病疫苗倡议（SATVI），在开发基于免疫的测定方面具有丰富的经验 分别用于候选HIV和结核病疫苗试验。到资金期结束时，我们将 合格的一套测定法，将补充现有方法来识别保护性的相关性 全细胞分枝杆菌疫苗效率研究的免疫力。