Hallmarks of Protective Immunity in Sequential Rhinovirus Infections in Humans

人类连续鼻病毒感染的保护性免疫标志

• 批准号: 9283305
• 负责人: Ronald B. Turner
• 金额: $ 89.29万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-06-01 至 2021-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9283305
• 关键词: Acute; Address; Adult; Antibodies; Antibody Formation; Antigenic Variation; B-Lymphocytes; BLR1 gene; Biochemistry; CD4 Positive T Lymphocytes; CD8-Positive T-Lymphocytes; Capsid Proteins; Cell Lineage; Cells; Chronic; Common Cold; Convalescence; Coupled; Cytometry; Development; Disease; Economic Burden; Economics; Effector Cell; Epitopes; Eragrostis; Evolution; Exposure to; Frequencies; Gene Expression Profiling; Health; Heterogeneity; Heterophile Antibodies; Human; Immune response; Immunity; Immunology; Immunophenotyping; Infection; Knowledge; Lung diseases; Maintenance; Mediating; Memory; Modeling; Molecular; Molecular Profiling; Molecular Target; Monitor; Morbidity - disease rate; Natural Killer Cells; Nature; Outcome; Peptides; Phase; Phenotype; Population; Production; Respiration Disorders; Rhinovirus; Role; Serotyping; Serum; Societies; Systems Biology; T cell response; T memory cell; T-Lymphocyte; Technology; Testing; Vaccine Design; Vaccines; Viral; Viral Physiology; Virus; Virus Shedding; Work; adaptive immunity; cell type; cellular targeting; computerized tools; cross reactivity; cytokine; design; health economics; high dimensionality; in vitro Assay; in vivo; longitudinal analysis; memory CD4 T lymphocyte; mortality; novel; prevent; response; single cell analysis; tool; transcriptomics; virology

SUMMARY We aim to understand the nature of CD4+ T cells that protect against rhinovirus (RV) infection. Rhinovirus infection is a major cause of common cold and an important trigger of disease exacerbations among those suffering from chronic lung disorders, thereby presenting an enormous health burden to society. The knowledge gained in this project could inform the design of vaccines to prevent repeat infections and their adverse sequelae, thereby greatly impacting human health. A major challenge is to identify those CD4+ T cells that protect against the numerous different strains of RV. Our preliminary findings suggest that circulating RV- specific CD4+ T cells directed against highly conserved regions of RV capsid proteins (VP1 and VP2) protect against infection. These cells comprise a mixture of memory effector T-cell (Teff, CXCR5neg) and T follicular helper (Tfh, CXCR5+) populations that we predict will mediate viral clearance (Teff), and promote antibody production by B cells (Tfh). Here, we address the hypothesis that RV-specific Teff and Tfh cells recognizing conserved RV epitopes confer cross-protection against RV strains through maintenance of stable lineage commitment and function. In Aim 1, we will confirm that virus-specific Teff and Tfh cells induced during the effector phase of RV infection persist into the memory phase and maintain their T-cell lineage relationships. To accomplish this objective, novel MHCII tetramers displaying conserved RV-16 peptides will be used to monitor the emergence and evolution of circulating RV-16-specific T cells in healthy adults who receive experimental challenge with RV-16. High-dimensional immunophenotyping will be performed using mass cytometry coupled with advanced computational tools to visualize and track discrete RV-specific T-cell populations, as well as synchronized fluxes in B-cell populations and other cell types including CD8+ T cells, T cells, NK cells, and NKT cells. Gene expression profiling, including single-cell transcriptomics, will be integrated to test whether the degree of homogeneity within RV-specific T-cell populations is preserved into the memory phase. In Aim 2, the capacity for pre-existing RV-16-specific T-cell populations induced by primary exposure to protect against re-infection with the heterotypic strain RV-39, will be addressed using a novel sequential RV challenge model that involves re-challenge with either RV-16 or RV-39 four months after RV-16 exposure. The relationship between higher numbers of pre-existing RV-16-specific T cells and correlates of protection against RV-39, including quantitative viral shedding and production of cross-reactive serum antibodies, will be determined. Further, we will explore how homotypic and heterotypic secondary viral exposures modulate T-cell populations with protective attributes. Finally, using in vitro assays, we will confirm that Teff and Tfh cells that persist after primary and secondary RV exposures can exert cross-protective anti-viral functions. By applying a single-cell systems biology approach, the outcomes are expected to provide the groundwork for the design of T-cell vaccines for RV, and to identify new cellular and molecular targets that might be exploited for treatment.

概括 我们旨在了解预防鼻病毒（RV）感染的CD4+ T细胞的性质。鼻病毒 感染是普通感冒的主要原因，也是疾病加剧的重要触发因素 患有慢性肺部疾病，从而给社会带来了巨大的健康负担。这 该项目获得的知识可以告知疫苗的设计，以防止重复感染及其 不良后遗症，从而极大地影响了人类健康。一个主要的挑战是识别那些CD4+ T细胞 防止众多不同的RV菌株。我们的初步发现表明循环RV- 针对高度保守的RV衣壳蛋白（VP1和VP2）的特定CD4+ T细胞保护 反对感染。这些细胞包含记忆效应的T细胞（TEFF，CXCR5NEG）和T卵泡的混合物 我们预测的助手（TFH，CXCR5+）种群将介导病毒清除率（TEFF），并促进抗体 B细胞（TFH）生产。在这里，我们解决了RV特异性TEFF和TFH细胞识别的假设 保守的RV表位通过维持稳定血统对RV菌株赋予交叉保护 承诺和功能。在AIM 1中，我们将确认特异性的TEFF和TFH细胞在 RV感染的效应阶段持续到记忆阶段并保持其T细胞谱系关系。到 完成这个显示保守RV-16肽的新型MHCII四聚体，用于监测 接受实验的健康成年人中循环RV-16特异性T细胞的出现和演变 RV-16的挑战。将使用质量细胞术耦合进行高维的免疫表型 使用先进的计算工具来可视化和跟踪离散的RV特异性T细胞群体以及 B细胞种群和其他细胞类型的同步通量，包括CD8+ T细胞，T细胞，NK细胞和 NKT细胞。基因表达谱分析，包括单细胞转录组学，将集成以测试是否是否 RV特异性T细胞种群中的同质性程度被保留到记忆阶段。在AIM 2中， 主要暴露引起的预先存在的RV-16特异性T细胞种群的能力以防止 将使用新型的顺序RV挑战模型来解决与异型应变RV-39的重新感染 这涉及RV-16暴露四个月后与RV-16或RV-39进行重新挑战。关系 在较高数量的现有RV-16特异性T细胞和针对RV-39的保护相关之间 将确定包括定量病毒脱落和交叉反应性血清抗体的产生。 此外，我们将探讨同型和异型次级病毒暴露如何调节T细胞种群 具有保护性属性。最后，使用体外测定，我们将确认在 一级和次级RV暴露可以发挥交叉保护的抗病毒功能。通过应用单细胞 系统生物学方法，预计结果将为T细胞的设计提供基础 RV的疫苗，并确定可能被利用用于治疗的新细胞和分子靶标。