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

各种纳米粒子（NP）已被用于递送小抗原，其病毒模拟物有限 然而，这些传统抗原在刺激 B 细胞免疫方面比可溶性抗原更有效。 NPs缺乏病毒“尖刺衣壳蛋白聚合物”的特征，例如聚合物上的尖刺抗原簇， 抗原簇之间的最佳距离以及尖峰上高度局部化的抗原密度如何实现尚不清楚。 传统纳米粒子缺乏病毒样特征，会影响 B 细胞免疫和持久的抗体反应。 尽管 B 细胞疫苗的病毒样特征可实现持久的 B 细胞免疫，但已使用病毒样特征进行了临床验证。 病毒衣壳蛋白颗粒（VLP），VLP不适合递送非衣壳小抗原（例如 细菌毒素、小分子和致癌肽），因为这些非衣壳小抗原不能 自组装成 VLP 需要开发用于小抗原的病毒样纳米颗粒来激活 B 细胞。 针对致命细菌毒素（炭疽、肉毒杆菌）、小分子和致癌肽的免疫力。 B 细胞免疫的三个组成部分对于持久的抗体反应至关重要： (A) 高效抗原 引流淋巴结中 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) 多价结合/交联激活抗原特异性 B 细胞， (C) 滤泡辅助 T 细胞 (Tfh) 的激活，支持生发中心 (GC) B 细胞及其分化 然而，目前尚不清楚 NPs 抗原缺乏病毒样特征是如何产生的。 递送系统影响 B 细胞免疫的这三个关键组成部分，以实现持久的抗体反应。 在这个提案中，我们将生成具有尖刺聚合物三个特征的无机病毒样纳米颗粒（IVLN） 使用四种小抗原（炭疽和肉毒毒素肽、小分子 4-羟基- 3-硝基苯基乙酰半抗原，HER2 肽）来检验我们的假设：(A) 病毒样特征。 IVLN 通过独特的抗原分布模式增强 B 细胞采集的有效递送/保留 淋巴结，(B) IVLN 的病毒样特征通过与 B 细胞多价结合/交联增强 B 细胞活化 受体，促进滤泡 T (Tfh) 细胞依赖性 B 细胞活化，增强长寿浆细胞的形成 （LLPC）位于生发中心（GC），并产生具有高特异性/亲和力的抗体，（C）病毒样特征 IVLN 诱导针对细菌毒素（炭疽和肉毒杆菌）和致癌物质的持久抗体反应 抗原。 目标 1 确定 IVLN 的病毒样特征，以改善独特抗原的抗原递送/保留 与传统 NP 相比，淋巴结中 B 细胞获取的分布模式 目标 2 通过 IVLN 与传统 NP 的病毒样特征来识别 B 细胞反应的阶段 目标 3 研究 IVLN 抗原免疫以诱导针对炭疽和炭疽的更持久的抗体反应 动物体内肉毒杆菌毒素和致癌 HER2 与传统 NP 的比较