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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.

项目摘要增强抗原性、抗体亲和力成熟和响应亚单位的记忆诱导的策略疫苗对于设计针对传染病的有效疫苗具有广泛的相关性，对于难以中和的病原体如HIV尤其重要。一种提高亚单位疫苗是以多价纳米颗粒形式配制抗原，这促进了几个方面的发展。的体液免疫，并最显着地增强B细胞受体（BCR）的交联。这种方法有已经在许可的疫苗（例如，HPV和HBV疫苗），以及各种疫苗临床前和临床开发。然而，到目前为止，仍然不清楚什么是理想的特性，纳米颗粒抗原展示。在这个项目中，我们使用独特的支架DNA折纸技术， 10-100纳米尺度的纳米颗粒，能够研究支架尺寸的影响，抗原拷贝数可达100以上，抗原-BCR亲和力，以及纳米级的空间组织和抗原呈递对BCR活化的维度。具体来说，我们测试的相对重要性，这些参数对B细胞活化，这是一个成功的亚基的发展至关重要疫苗，使用称为eOD-GT 8的HIV-1 gp 120的生殖系靶向工程化外结构域，及其具有不同亲和力的变体，作为测试平台。早期B细胞信号传导和途径活化的体外评价将被表征，并与基准强烈激活60聚体控制组织在一个蛋白质支架单细胞荧光成像用于研究BCR结合的详细机制和B细胞活化的基础上发现的最佳免疫原呈递。这些构建体，然后用于使用小鼠，测试这些最佳HIV DNA-NP构建体对体内T细胞和B细胞应答的影响模型总之，我们的结果将提供最佳免疫原呈递参数的阐明在开发更有效的亚单位疫苗时，有效的免疫细胞应答，主要翻译艾滋病毒和其他传染病的可能性。