Investigating the MHC Class II-Restricted Processing Landscape of HIV-1 Antigens

研究 HIV-1 抗原的 MHC II 类限制加工景观

• 批准号: 10004493
• 负责人: Mary Margaret Addison
• 金额: $ 3.27万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-01 至 2021-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10004493
• 关键词: Acquired Immunodeficiency Syndrome; Adaptive Immune System; Address; Antigen Presentation; Antigen Presentation Pathway; Antigen-Presenting Cells; Antigens; B-Lymphocytes; Binding; Biological Assay; Biomedical Research; Blood; CD4 Positive T Lymphocytes; CD8-Positive T-Lymphocytes; CD86 gene; Cells; Cellular biology; Chemicals; Communication; Complex; Cross Presentation; Data; Dendritic Cells; Detection; Development; Early Endosome; Epitopes; Fellowship; Foundations; Future; Generations; Goals; HIV; HIV Antigens; HIV Infections; HIV vaccine; HIV-1; Heterogeneity; Histocompatibility Antigens Class II; Human; Immunology; In Vitro; Infection; Influenza; Knowledge; Laboratories; Lead; Literature; MHC Class II Genes; Pathway interactions; Pattern; Peptides; Physiological; Play; Population; Predisposition; Process; Production; Recycling; Reporting; Research; Research Personnel; Role; Scientist; Senior Scientist; Small Interfering RNA; Standard Model; Surface; Surface Antigens; System; T cell response; T-Cell Activation; Technical Expertise; Testing; Training; Vaccine Design; Vaccines; Viral; Viral Antigens; Viral Load result; Viremia; Virion; Virus; Work; adaptive immune response; anergy; antigen processing; base; cell type; design; improved; in vivo; influenzavirus; inhibitor/antagonist; insight; macrophage; monocyte; neutralizing antibody; pathogen; receptor; response; skills

Project Summary Robust HIV-specific CD4 T cell responses are associated with decreased viral load, an increased antibody neutralization breadth, and a slower progression to AIDS, and should therefore be considered in the development of a protective HIV vaccine. CD4 T cells are activated by antigen-derived peptides (epitopes) displayed in complex with MHC Class II molecules (MHCII) on the surface of an antigen presenting cell (APC). According to the standard model, internalized antigens are catabolized (“processed”) and loaded onto MHCII in late endosomal compartments. However, many alternative pathways have been described. These include the recycling pathway, in which peptide is loaded onto MHCII in the early endosome, and endogenous processing, which occurs when virally-derived antigens synthesized within infected APCs are proteolyzed and loaded onto MHCII via a complex network of intracellular pathways. While some HIV-1 epitopes can be presented from inactivated virus, others have been shown to be presented endogenously. However, the relative contributions of classical and alternative pathways to the HIV-specific CD4 T cell response are not known. Additionally, our understanding of the cell types that act as APCs during HIV-1 infection, and their relative contributions to CD4 T cell activation, is incomplete. Of note, in addition to the “professional” APCs that present peptide via MHCII, CD4 T cells, which transiently express MHCII upon stimulation and are host cells for HIV, might act in this capacity. Accordingly, I will investigate the abilities of human dendritic cells, macrophages, and CD4 T cells to present HIV-1 epitopes from infectious and inactivated virus, and elucidate the cellular pathways utilized by these APCs for peptide production and MHCII loading. In Aim 1, I will define the differential abilities of dendritic cells, macrophages, and CD4 T cells to present epitopes from live and inactivated HIV-1 using established in vitro antigen presentation assays. In Aim 2, I will elucidate the cellular mechanisms used by DCs, macrophages, and CD4 T cells to present HIV antigens via MHC-II using siRNA and chemical inhibitors targeting key aspects of the antigen processing machinery. This work will provide insights into the pathways used by APCs to activate CD4 T cells in the context of an HIV-1 infection and has the potential to inform rational vaccine design. This project will allow me to develop into a confident, independent researcher. This training will expand my knowledge of cell biology and immunology and help me master valuable new technical skills. Additionally, I will improve my communication skills, allowing me to effectively communicate the importance of biomedical research with the public. Most importantly, this training will help me achieve my long- term professional goal of being an independent senior scientist.

项目摘要 稳健的HIV特异性CD 4 T细胞应答与病毒载量降低、抗体水平升高相关 中和宽度，和较慢的进展，艾滋病，因此，应考虑在 开发保护性艾滋病毒疫苗。CD 4 T细胞被抗原衍生肽（表位）激活 与抗原呈递细胞（APC）表面上的MHC II类分子（MHCII）复合展示。 根据标准模型，内化的抗原被分解代谢（“加工”）并加载到MHCII上， 晚期内体隔室然而，已经描述了许多替代途径。其中包括 再循环途径，其中肽在早期内体中装载到MHCII上，以及内源性加工， 当在感染的APC内合成的病毒衍生抗原被蛋白水解并装载到 MHCII通过复杂的细胞内途径网络。虽然一些HIV-1表位可以从 灭活病毒，其他已被证明是内源性的。相对贡献 HIV特异性CD 4 T细胞应答的经典和替代途径尚不清楚。此外，我们的 了解在HIV-1感染过程中充当APC的细胞类型，以及它们对CD 4+细胞的相对贡献。 T细胞活化不完全。值得注意的是，除了通过MHCII呈递肽的“专业”APC外， CD 4 T细胞在刺激时瞬时表达MHCII，是HIV的宿主细胞，可能在这一过程中起作用。 容量因此，我将研究人类树突状细胞，巨噬细胞和CD 4 T细胞的能力， 目前的HIV-1表位从传染性和灭活病毒，并阐明细胞利用的途径， 这些APC用于肽生产和MHCII装载。在目标1中，我将定义 树突状细胞、巨噬细胞和CD 4 T细胞呈递来自活的和灭活的HIV-1的表位， 建立了体外抗原呈递试验。在目标2中，我将阐明 使用siRNA和化学抑制剂通过MHC-II呈递HIV抗原的DC、巨噬细胞和CD 4 T细胞 靶向抗原加工机制的关键方面。这项工作将提供深入了解的途径 在HIV-1感染的背景下，APC用于激活CD 4 T细胞，并有可能告知 合理的疫苗设计这个项目将使我成为一个自信，独立的研究人员。这 培训将扩展我的细胞生物学和免疫学知识，并帮助我掌握有价值的新技术。 skills.此外，我将提高我的沟通技巧，使我能够有效地沟通， 生物医学研究的重要性。最重要的是，这次培训将帮助我实现我的长期- 长期的职业目标是成为一名独立的资深科学家。