Virus-Like Nanoparticles for Non-Capsid Antigen Delivery with Virus Structure/Functional Mimicry to Activate B Cell Immunity

用于非衣壳抗原递送的病毒样纳米颗粒，具有病毒结构/功能拟态以激活 B 细胞免疫

• 批准号: 10212209
• 负责人: Irina Leonidovna Grigorova
• 金额: $ 51.32万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-07 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10212209
• 关键词: Affect; Affinity; Animals; Anthrax disease; Antibodies; Antibody Formation; Antibody Response; Antibody Specificity; Antibody titer measurement; Antigens; B-Cell Activation; B-Cell Antigen Receptor; B-Lymphocytes; Bacterial Infections; Bacterial Toxins; Binding; Botulinum Toxins; Capsid Proteins; Clinical; Development; ERBB2 gene; Engineering; Exhibits; Haptens; Helper-Inducer T-Lymphocyte; Immunity; Immunization; Malignant Neoplasms; Memory B-Lymphocyte; Oncogenic; Pattern; Peptides; Plasma Cells; Prevention; Specificity; Structure; Structure of germinal center of lymph node; Supporting Cell; System; T-Lymphocyte; Testing; Vaccines; Viral; Virus; Virus-like particle; anthrax toxin; botulinum; cancer therapy; crosslink; density; draining lymph node; improved; lymph nodes; mimicry; nanoparticle; nanoparticle delivery; prevent; response; small molecule; virus characteristic

Various nanoparticles (NPs) have been used for delivery of small antigens, which have limited viral mimic features and are more efficacious than soluble antigens in stimulating B-cell immunity. However, these traditional NPs lack characteristics of virus “spiky capsid protein peplomer”, e.g. spiky antigen clusters on the peplomers, optimal distance between antigen clusters, and highly localized antigen density on the spike. It is unknown how the lack of virus-like features of traditional NPs affect B cell immunity and durable antibody responses. Although virus-like features of B cell vaccine for durable B cell immunity are clinically validated using virus-like particles (VLPs) of viral capsid proteins, VLPs are not suitable for delivery of non-capsid small antigens (such as bacterial toxins, small molecules, and oncogenic peptides) since these non-capsid small antigens are not able to self-assemble to VLPs. There is a need to develop virus-like nanoparticles for small antigens to activate B cell immunity against deadly bacterial toxins (Anthrax, Botulinum), small molecules, and oncogenic peptides. Three components of B cell immunity are critical for durable antibody response: (A) Efficient antigen delivery/retention and unique antigen distribution patterns for B cell acquisition in the draining lymph nodes (dLNs), (B) Activation of antigen-specific B cells through multivalent binding/crosslink with B cell receptor (BCR), (C) Activation of follicular T Helper cells (Tfh) that support Germinal Center (GC) B cells and their differentiation to long-lived plasma cells (LLPCs). However, it is unknown how the lack of virus-like features of NPs antigen delivery systems affect these three critical components of B cell immunity for durable antibody response. In this proposal, we will generate inorganic virus like nanoparticles (IVLNs) with three features of spiky peplomers of virus' using four types of small antigens (peptides of anthrax and botulinum toxins, small molecule 4-hydroxy- 3-nitrophenyl acetyl-hapten, HER2 peptides) to test our hypothesis. We hypothesize that: (A) Virus-like features of IVLNs enhance efficient delivery/retention with unique antigen distribution patterns for B cell acquisition in the lymph node, (B) Virus-like features of IVLNs enhance B cell activation via multivalent bind/crosslink with B cell receptor, promote follicular T (Tfh) cell-dependent B-cell activation, enhance formation of long-lived plasma cells (LLPCs) in the Germinal Center (GC), and generate antibodies with high specificity/affinity, (C) Virus-like features of IVLNs induce durable antibody response against bacterial toxins (anthrax and botulinum) and oncogenic antigens. Aim 1 Determine virus-like features of IVLNs to improve antigen delivery/retention with unique antigen distribution patterns for B cell acquisition in the lymph nodes vs. traditional NPs Aim 2 Identify the stages of B cell responses by the virus-like features of IVLNs vs. traditional NPs Aim 3 Investigate IVLNs-antigen immunizations to induce more durable antibody response against Anthrax and Botulinum toxins and oncogenic HER2 in animals vs. traditional NPs

各种纳米颗粒（NP）已用于递送小抗原，这些抗原的病毒模拟有限 特征，比固体抗原在刺激B细胞免疫方面更有效。但是，这些传统 NPs缺乏病毒“尖状的衣壳蛋白化合物”的特征，例如胡椒体上的尖峰抗原簇， 抗原簇之间的最佳距离和尖峰上高度局部的抗原密度。这是未知的 传统NP缺乏类似病毒的特征会影响B细胞免疫和耐用的抗体反应。 尽管使用病毒样的B细胞疫苗的病毒样特征在临床上验证 病毒式衣壳蛋白的颗粒（VLP），VLP不适合递送非capsid小抗原（例如 细菌毒素，小分子和致癌胡椒体），因为这些非粘蛋白小抗原无法 自我组装到VLP。需要开发类似病毒的纳米颗粒来激活B细胞 对致命细菌毒素（炭疽，肉毒杆菌），小分子和致癌肽的免疫力。 B细胞免疫的三个成分对于耐用抗体反应至关重要：（a）有效的抗原 排放淋巴结中B细胞获取的递送/保留和独特的抗原分布模式 （DLNS），（B）通过与B细胞受体（BCR）的多价结合/交联，激活抗原特异性B细胞， （c）支持生发中心（GC）B细胞及其分化的卵泡T辅助细胞（TFH）的激活 到长寿命的浆细胞（LLPC）。但是，尚不清楚如何缺乏NPS抗原的病毒样特征 递送系统影响B细胞免疫的这三个关键成分，以实现耐用抗体反应。 在此提案中，我们将生成具有三种尖刺胡椒体特征的纳米颗粒（IVLN）等无机病毒 使用四种类型的小抗原（炭疽和肉毒杆菌毒素的肽，小分子4-羟基 - 3-亚硝基苯基乙酰基，HER2肽）以检验我们的假设。我们假设：（a）类似病毒的特征 IVLN的IVLN可增强有效的递送/保留率，并具有独特的抗原分布模式，用于B细胞获取 淋巴结，（b）IVLN的病毒样特征通过与B细胞的多价结合/交联增强B细胞激活 受体，促进卵泡T（TFH）细胞依赖性B细胞活化，增强长寿命的浆细胞的形成 （LLPC）在生发中心（GC）中，并生成具有高特异性/亲和力的抗体，（c）病毒样特征 IVLN的诱导抗细菌毒素（炭疽和肉毒杆菌）和致癌的耐用抗体反应 抗原。 AIM 1确定IVLN类似病毒的特征，以改善独特的抗原的抗原递送/保留率 淋巴结与传统NP中B细胞获取的分布模式 AIM 2通过IVLN与传统NP的病毒样特征确定B细胞反应的阶段 AIM 3研究IVLNS-抗原免疫抑制剂，以诱导针对炭疽和的耐用抗体反应 动物与传统NP的肉毒杆菌毒素和致癌HER2