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：
10253355
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/10253355
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 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 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 Investigation Ligands Memory Modeling Modification Nanotechnology Peripheral Receptor Activation Receptor Signaling Resolution Role Scaffolding Protein Signal Pathway Signal Transduction Stimulus Structure Subunit Vaccines Surface T-Lymphocyte Technology Testing Toll-like receptors Vaccination Vaccine Design Vaccines Variant Viral Virus-like particle adaptive immune response base 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 vaccine development

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，抗体亲和力成熟和记忆诱导的策略，以响应亚基疫苗对于针对感染性疾病的有效疫苗的设计具有广泛的意义，并且可能是对于难以中和病原体（例如HIV）尤其重要。一种提高效率的方法亚基疫苗是在多价，纳米含量的形式中构造抗原，该形式促进了几个方面体液免疫力，最著名的是增强了B细胞受体（BCR）的交联。这种方法有在有执照的疫苗（例如HPV和HBV疫苗）和各种疫苗中探索在临床前和临床发展中。但是，迄今为止尚不清楚什么是理想特征纳米粒子抗原显示。在这个项目中，我们使用脚手架DNA折纸的独特技术来工程师 10–100纳米尺度上的纳米颗粒，可研究脚手架大小的影响，抗原拷贝数高达100多个抗原-BCR亲和力以及纳米级空间组织抗原激活中抗原表现的维度。具体来说，我们测试了这些关于B细胞激活的参数，这些参数对于成功的亚基的开发至关重要使用靶向HIV-1 GP120的工程外部域的种系，称为EOD-GT8，其疫苗称为EOD-GT8及其具有不同亲和力的变体作为测试床。早期B细胞信号传导和途径激活的体外评估将被表征，并与基准强烈激活60-mer对照的基准形成鲜明对比。蛋白质支架。单细胞荧光成像用于研究BCR结合的详细机制基于发现的最佳免疫呈现的B细胞激活。然后将这些结构用于使用小鼠在体内测试这些最佳HIV DNA-NP构建体对T细胞和B细胞反应的影响型号。综上所述，我们的结果将提供最佳免疫呈现参数的阐明在开发更有效的亚基疫苗中，有效的免疫细胞反应，并进行了主要的转化艾滋病毒和其他传染病的潜力。