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

基本信息

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
项目类别：
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

项目摘要

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.

项目概要（摘要） 2013 年，结核分枝杆菌 (M.tb) 导致了超过 150 万人死亡，这使得目前，牛分枝杆菌卡介苗 (BCG) 是全球主要的传染性死亡原因。卡介苗是唯一获得许可的结核病疫苗，可预防结核病的传播。儿童疾病，但在预防成人肺结核方面效果不一致。成人肺结核传染性强，卡介苗无法控制疫情，迫切需要新疫苗目前有多种疫苗正在开发中，其中包括。重组 BCG 和减毒 M.tb 菌株对 M.tb 的成功免疫反应主要取决于 T-。细胞，通常被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，候选全细胞疫苗是多抗原的，并且含有肽和人类 T 细胞识别的非肽抗原已最终确定。研究表明，当与抗原呈递细胞上的 CD1 蛋白结合时，可以激活人类 T 细胞。该系统在哺乳动物中是保守的，并且在其他脊椎动物中大多不存在，这表明它进化到执行与 MHC 系统非冗余的重要功能，但尚未经过验证。我们拥有测量人类 T 细胞针对非肽抗原（例如脂质）反应的测定方法。与脂质抗原发现、流式细胞术、疫苗和计算领域的专家建立了合作伙伴关系我们最近开发了可溶性 CD1，以应对过去五年的这一技术挑战。四聚体和基于激活的 T 细胞分析测定作为促进基于群体的 T 细胞研究的工具我们还开发了对分枝杆菌脂质的反应的无偏计算方法。在目标 1 中，我们将优化并鉴定使用脂质的测定。加载 CD1 四聚体来研究静止状态下脂质特异性 T 细胞的记忆表型和激活状态。目标2，我们将优化和限定基于互补激活的T测定的效应功能研究与 MHC 蛋白不同，CD1 蛋白实际上是非多态性的，因此这些细胞被脂质抗原激活。在目标 3 中，我们将使用这两种检测来研究遗传背景。使用 BCG 作为模型系统，研究分枝杆菌疫苗接种对人类脂质特异性 T 细胞反应的影响。该提案的主要合作者是 HIV 疫苗试验网络 (HVTN) 和南非结核疫苗倡议 (SATVI)，在开发基于免疫的检测方法方面拥有丰富的经验到资助期结束时，我们将分别进行候选艾滋病毒和结核病疫苗试验。鉴定了一套检测方法，将补充现有的方法来识别保护性的相关性全细胞分枝杆菌疫苗功效研究中的免疫。