Lymph node targeting nanoparticles for HIV Env proteins

项目总结 随着非常成功的抗逆转录病毒治疗的到来，与人类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，数十万 每年都会出现新的艾滋病毒感染病例，这表明需要一种安全有效的疫苗。HIV疫苗 基于广谱中和抗体(BNAbs)的激发，是艾滋病毒研究的前沿。稳定下来 HIV免疫原现在已经显示出在恒河猴体内诱导二级中和抗体的能力，并且 已从人类受试者中分离出大量的bNAb。此外，生殖系靶向免疫原具有 证明了在人类中初步成功地诱导了bNAb生殖系前体。有效性的一个关键因素 艾滋病毒免疫原的一种传递方式。体细胞超突变是实现最佳和高效的关键事件 BNab启发式。重组HIV env蛋白免疫原性差，不能跟随典型的淋巴结节 (Ln)由于高糖基化和天然IgM识别能力差而导致的转运途径。相反，重组 HIV env蛋白被滤泡间巨噬细胞捕获，而不是被包膜下的巨噬细胞捕获 窦，导致抗原可利用性差，并限制重复的体细胞过度突变。科学的前提是 因为这个提议是因为重组HIV env蛋白改变了LN的运输，可生物降解的LN 靶向纳米颗粒有助于将可溶的HIV环境三聚体直接转移到LNS，是一种很有前途的工具 绕过当前的障碍。我们的目标是验证这样一种假设，即靶向纳米颗粒的淋巴结膜 (NP)技术将改善人类免疫缺陷病毒(HIV)包膜产生的免疫反应 (Env)蛋白质免疫原。这一提议风险很高，因为它会引发广泛的中和抗体。 (BNAbs)滴度高、持续时间长是具有挑战性的；然而，如果成功，它将通过 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-PEGNPs 更有效地将种系靶向免疫原输送到LN毛囊并增加体细胞超突变 与未包封的免疫原相比。将用种系靶向免疫原三聚体免疫小鼠 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的研究。LN在卵泡中的转运和积累 也将接受评估。在具体目标2中，我们将确定在NP中是否共封装不同的HIV env 增强第二层中和广度的启发。来自两个病毒分支的HIV Env SOSIP三聚体将被组合 在单个NP中，将评估血清学反应。这项建议是高度协作的，结合了Dr。 结构免疫学家、艾滋病毒/艾滋病疫苗学者计划获得者Jarrod Mousa和海权博士 他是一位生物材料和纳米医学专家。我们的发现具有显著改善艾滋病毒环境的巨大潜力 我们的发现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.