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

不同的纳米颗粒(Np)已经被用来传递小的抗原，它们具有有限的病毒模拟性。 在刺激B细胞免疫方面比可溶性抗原更有效。然而，这些传统的 NPS缺乏病毒“刺状衣壳蛋白聚合体”的特征，例如聚集物上的刺状抗原团， 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。目前还不清楚是如何做到的 传统NPs缺乏类似病毒的特性，影响B细胞免疫和持久的抗体反应。 尽管用于持久B细胞免疫的B细胞疫苗的病毒样特征在临床上是用类病毒来验证的 病毒衣壳蛋白的颗粒(VLP)，VLP不适合运送非衣壳小抗原(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。需要开发针对小抗原的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细胞免疫的三个组成部分对持久的抗体反应至关重要：(A)有效的抗原 B细胞在引流淋巴结中的传递/滞留和独特的抗原分布模式 (B)通过与B细胞受体(BCR)的多价结合/交联来激活抗原特异性B细胞； (C)支持生发中心(GC)B细胞的滤泡辅助性T细胞(TFH)的激活及其分化 到长寿命的浆细胞(LLPC)。然而，目前尚不清楚NPs抗原缺乏病毒样特征的原因 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。 在这项提议中，我们将产生具有尖峰聚合体三个特征的无机病毒样纳米颗粒(IVLN) 使用四种类型的小抗原(炭疽和肉毒杆菌毒素多肽，小分子4-羟基- 3-硝基苯乙酰-半抗原，HER2肽)来验证我们的假设。我们假设：(A)类似病毒的特征 的IVLN通过独特的抗原分布模式增强B细胞在体内的有效传递/保留 淋巴转移，(B)IVLN的病毒样特征通过与B细胞的多价结合/交联增强B细胞的激活 受体，促进滤泡T(TFH)细胞依赖的B细胞活化，促进长寿浆细胞的形成 (C)病毒样特征 诱导对细菌毒素(炭疽和肉毒杆菌)和致癌的持久抗体反应 抗原。 目的1确定IVLN的病毒样特征，以改善独特抗原的抗原传递/保留 B细胞在淋巴结中的分布模式与传统NPs的比较 目的2通过IVLN与传统NPs的病毒样特征识别B细胞应答的阶段 目的研究免疫球蛋白疫苗诱导更持久的抗炭疽抗体应答。 肉毒杆菌毒素和致癌HER2在动物和传统NPs中的比较

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