Lymph node targeting nanoparticles for HIV Env proteins

项目摘要 随着非常成功的抗逆转录病毒疗法的出现，与人类疾病相关的死亡率和发病率 免疫缺陷病毒（HIV）感染已显著减少。尽管如此， 每年都有新的艾滋病毒感染病例发生，这表明需要一种安全有效的疫苗。艾滋病毒疫苗 基于广泛中和抗体（bNAb）的启发，是艾滋病研究的前沿。稳定 HIV免疫原现在已经显示出在恒河猴中引发2级中和抗体的能力， 已经从人类受试者中分离出许多bNAb。此外，生殖系靶向免疫原具有 在人类中初步成功诱导bNAb生殖系前体。有效性的一个关键方面是 艾滋病免疫原的传递方式。体细胞超突变是优化和有效的 bNAb诱导。重组HIV Env蛋白免疫原性差，不能跟随典型的淋巴结 (LN)由于高糖基化和天然IgM识别不良，导致运输途径。相反，重组 HIV Env蛋白被滤泡间巨噬细胞捕获，而不是被包膜下巨噬细胞捕获 窦，导致抗原可用性差和有限的重复体细胞超突变。科学前提 由于重组HIV Env蛋白改变了LN的运输， 促进可溶性HIV Env三聚体直接转移到LN的靶向纳米颗粒是一种有前途的工具， 突破现有壁垒。我们的目的是检验淋巴结（LN）靶向纳米颗粒 (NP)技术将改善人类免疫缺陷病毒（HIV）包膜产生免疫反应 (Env)蛋白免疫原。这一建议是高风险的，因为广泛中和抗体的激发 高滴度和长持续时间的bNAbs是具有挑战性的;尽管如此，如果成功，它提供了高回报， 大大提高了HIV Env疫苗的反应。在具体目标1中，我们将确定PLGA-b-PEG NP是否 更有效地将生殖系靶向免疫原递送至LN卵泡并增加体细胞超突变 与未封装的免疫原相比。小鼠将用生殖系靶向免疫原三聚体免疫 封装在NP平台内，并将完成血清学和细胞分析，包括监测 使用单细胞测序方法进行体细胞超突变。LN在卵泡中的运输和积聚 也将被评估。在具体目标2中，我们将确定是否将不同的HIV Env共包封在NP中， 提高了2级中和广度的诱导。来自两个病毒进化枝的HIV Env SOSIP三聚体将被合并 在单个NP中，将评估血清学应答。这项建议是高度合作，结合博士。 Jarrod Mousa，结构免疫学家和艾滋病毒/艾滋病疫苗学者计划的接受者，和Hai-Quan博士 生物材料和纳米医学专家毛。我们的研究结果有很大的潜力显着改善艾滋病毒的环境 我们的研究结果将适用于整个艾滋病毒疫苗领域。

PROJECT SUMMARY With the advent of highly successful antiretroviral therapy, mortality and morbidity associated with human immunodeficiency virus (HIV) infection has been significantly reduced. Despite this, hundreds of thousands of new HIV infections occur each year, demonstrating the need for a safe and effective vaccine. HIV vaccines based on the elicitation of broadly neutralizing antibodies (bNAbs) are at the forefront of HIV research. Stabilized HIV immunogens have now shown the ability to elicit tier 2 neutralizing antibodies in rhesus macaques, and numerous bNAbs have been isolated from human subjects. Furthermore, germline targeting immunogens have demonstrated initial success in humans in eliciting bNAb germline precursors. A key aspect in the effectiveness of HIV immunogens is the delivery method. Somatic hypermutation is a critical event for optimal and efficient bNAb elicitation. Recombinant HIV Env proteins are poorly immunogenic and fail to follow typical lymph node (LN) trafficking pathways due to high glycosylation and poor recognition by natural IgM. Instead, recombinant HIV Env proteins are captured by interfollicular macrophages rather than by macrophages in the subcapsular sinus, leading to poor antigen availability and limited repeated somatic hypermutation. The scientific premise for this proposal is that since recombinant HIV Env proteins have altered LN trafficking, biodegradable LN targeting nanoparticles that facilitate transfer of soluble HIV Env trimers directly to LNs are a promising tool to circumvent current barriers. Our objective is to test the hypothesis that a lymph node (LN) targeting nanoparticle (NP) technology will improve immune responses generated by human immunodeficiency virus (HIV) envelope (Env) protein immunogens. This proposal is high risk, as the elicitation of broadly neutralizing antibodies (bNAbs) at a high titer and long duration is challenging; nonetheless, if successful, it offers high reward by substantially improving HIV Env vaccine responses. In Specific Aim 1, we will determine if PLGA-b-PEG NPs more efficiently deliver germline targeting immunogens to LN follicles and increase somatic hypermutation compared to unencapsulated immunogens. Mice will be immunized with germline targeting immunogen trimers encapsulated within the NP platform, and serological and cellular analysis will be completed, including monitoring of somatic hypermutation using single cell sequencing approaches. LN trafficking and accumulation in follicles will also be assessed. In Specific Aim 2, we will determine if co-encapsulation of diverse HIV Envs in NPs enhances elicitation of tier 2 neutralization breadth. HIV Env SOSIP trimers from two viral clades will be combined in a single NP, and serological responses will be assessed. This proposal is highly collaborative, combining Dr. Jarrod Mousa, a structural immunologist and HIV/AIDS Vaccine Scholars Program recipient, and Dr. Hai-Quan Mao, a biomaterials and nanomedicine expert. Our findings have high potential to dramatically improve HIV Env responses, and our findings will be applicable to the entire HIV vaccine field.