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Investigation of Synthetic DNA-based Viral Particles for Spatially Controlled Antigen Presentation

基于 DNA 的合成病毒颗粒空间控制抗原呈递的研究

基本信息

批准号：
10662377
负责人：
Mark Bathe
金额：
$ 38.78万
依托单位：
MASSACHUSETTS INSTITUTE OF TECHNOLOGY
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-08-02 至 2025-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10662377
关键词：
3-Dimensional Acquired Immunodeficiency Syndrome Adjuvant Affinity Agonist Animal Model Antibody Affinity Antibody Formation Antigen Presentation Antigen-Presenting Cells Antigens Artificial nanoparticles B-Cell Activation B-Cell Antigen Receptor B-Cell Receptor Binding B-Lymphocytes Bathing Benchmarking Biodistribution Biological Assay Biomedical Engineering Calcium Signaling Cell model Cells Cellular Immunity Characteristics Communicable Diseases Confocal Microscopy Cues DNA Development Dimensions Engineering Ensure Evaluation Event HIV HIV Envelope Protein gp120 HIV Infections HIV envelope protein HIV vaccine HIV-1 Health Hepatitis B Vaccines Heterogeneity Human Human Papilloma Virus Vaccine Humoral Immunities Image Immobilization Immune Immune response Immunologic Adjuvants Immunology In Vitro Infection Innate Immune Response Investigation Licensing Ligands Memory Modeling Modification Nanotechnology Pathway interactions Peripheral Receptor Activation Receptor Signaling Role Scaffolding Protein Signal Transduction Stimulus Structure Subunit Vaccines Surface T-Lymphocyte Technology Testing Toll-like receptors Vaccination Vaccine Design Vaccines Variant Viral Virus-like particle Visualization adaptive immune response cellular imaging clinical development clinically relevant crosslink density design env Gene Products experience flexibility fluorescence imaging human model immune activation in vivo in vivo evaluation link protein lymph nodes mouse model nanocluster nanometer nanoparticle nanoparticulate nanoscale neutralizing antibody pandemic disease particle pathogen preclinical development preservation prevent response scaffold synthetic construct tool translational potential ultra high resolution vaccine development vaccine formulation

项目摘要

PROJECT SUMMARY Strategies to enhance antigenicity, antibody affinity maturation, and memory induction in response to subunit vaccines are of broad relevance for the design of effective vaccines against infectious diseases, and may be especially important for difficult-to-neutralize pathogens such as HIV. One approach to enhance the efficacy of subunit vaccines is to formulate antigens in a multivalent, nanoparticulate form, which promotes several aspects of humoral immunity, and most notably enhances crosslinking of B cell receptors (BCRs). This approach has been exploited both in licensed vaccines (e.g., the HPV and HBV vaccines), and in a great variety of vaccines in preclinical and clinical development. However, to date it remains unclear what are the ideal characteristics of nanoparticle antigen display. In this project, we use the unique technology of scaffolded DNA origami to engineer nanoparticles on the 10–100 nanometer scale that offer the ability to investigate the impact of scaffold size, antigen copy number up to more than 100, antigen-BCR affinity, as well as the nanoscale spatial organization and dimensionality of antigen presentation on BCR activation. Specifically, we test the relative importance of these parameters on B-cell activation, which are of central importance to the development of a successful subunit vaccines, using the germline targeting engineered outer domain of HIV-1 gp120, termed eOD-GT8, and its variants with different affinities, as a testbed. In vitro evaluation of early B-cell signaling and pathway activation will be characterized, and contrasted with the benchmark strongly activating 60-mer control organized on a protein scaffold. Single-cell fluorescence imaging is used to investigate the detailed mechanism of BCR-binding and B-cell activation based on the optimal immunogen presentation found. These constructs are then used to test the impact of these optimal HIV DNA-NP constructs on T-cell and B-cell response in vivo using mouse models. Taken together, our results will offer the elucidation of the optimal immunogen presentation parameters for effective immune cell response in the development of more effective subunit vaccines, with major translational potential for HIV and other infectious diseases.

项目总结对亚单位反应增强抗原性、抗体亲和力成熟和记忆诱导的策略疫苗对于设计有效的传染病疫苗具有广泛的相关性，并可能对艾滋病毒等难以中和的病原体尤其重要。提高药效的一种方法亚单位疫苗是以多价、纳米颗粒的形式形成抗原，这促进了几个方面增强体液免疫，最显著的是增强B细胞受体(BCR)的交联性。这种方法具有在获得许可的疫苗(例如，HPV和乙肝疫苗)和各种疫苗中都得到了利用在临床前和临床开发方面。然而，到目前为止，人们仍然不清楚什么是理想的特征纳米颗粒抗原展示。在这个项目中，我们使用了独特的支架DNA折纸技术来工程 10-100纳米尺度的纳米颗粒，能够研究支架尺寸的影响，抗原拷贝数高达100多个，抗原与bcr亲和力强，以及纳米尺度的空间组织 BCR活化的抗原提呈维度。具体地说，我们测试了这些关于B细胞激活的参数，对于成功的亚单位的发展是至关重要的使用种系靶向HIV-1 gp120工程外区的疫苗，称为EOD-GT8，及其不同亲和力的变种，如试验床。早期B细胞信号转导和通路激活的体外研究将被描述为特征，并与组织在蛋白质支架。用单细胞荧光成像技术研究BCR结合的详细机制以及基于找到的最佳免疫原提呈物激活B细胞。然后使用这些构造来用小鼠体内检测这些优化的HIV DNA-NP结构对T细胞和B细胞应答的影响模特们。综上所述，我们的结果将为最佳免疫原呈递参数的阐明提供依据。在开发更有效的亚单位疫苗中进行有效的免疫细胞反应，主要是翻译有可能感染艾滋病毒和其他传染病。