Design and characterization of biomimetic nanobiomaterials to elicit CD1-restricted T cell responses during sub-unit vaccination

项目摘要 亚单位疫苗将来自病原体的免疫显性蛋白或肽抗原与选择性抗原结合。 佐剂，旨在为减毒疫苗提供更可扩展、可重复、低成本和快速的替代品 含有活病原体的细菌不幸的是，目前的亚单位疫苗缺乏脂质抗原，并且很少达到预期的效果。 广泛的T细胞反应需要持久的免疫记忆和保护。相比之下， 疫苗缺乏定制性和可扩展性，但包含整个病原体以提供蛋白质和脂质 免疫过程中的抗原。这种脂质和蛋白质抗原的组合激活了广谱的 效应T细胞，包括常规的MHC限制性T细胞，其响应于肽并表现出相当大的 多态性，以及针对特定脂质的非多态性CD 1限制性T细胞。一个更 因此，同时激活脂质和肽特异性T细胞的仿生策略可能表明， 与仅限于蛋白抗原的亚单位疫苗相比，增强的效力和控制。 目前的亚单位疫苗和免疫疗法忽视脂质抗原主要是由于1） 脂质靶向递送的困难，和2）缺乏合适的小鼠模型。在人类中，CD 1家族 由第1组CD 1分子（CD 1a、CD 1b和CD 1c）和第2组CD 1分子CD 1d组成。老鼠， 而仅表达CD 1d。该项目涉及由领导的研究小组之间的密切合作， 生物工程师和基础免疫学家，旨在通过设计纳米生物材料来克服这些障碍， 脂质和蛋白质抗原与佐剂组合的增强的双重递送， 人源化CD 1转基因（hCD 1 Tg）小鼠中的MHC限制性T细胞应答。为了表征、优化和 将这些新型纳米生物材料与世界上最常用的减毒疫苗进行比较， 卡介苗（BCG）结核病（TB）疫苗，提出了以下目的：在目的1中，体外 体内方法将确定最佳的纳米生物材料和佐剂组合， 联合的CD 1和MHC限制性T细胞应答。在目标2中，将使用脂质/蛋白质多抗原方法。 在用毒性结核分枝杆菌（Mtb）攻击的hCD 1 Tg小鼠中验证。在目标3中，一种新型水凝胶 载脂抗原纳米生物材料的控释和缓释将采用递送系统 评估慢性CD 1限制性T细胞活化的有效性和安全性。这项拟议中的研究将提供一个“证据 将Mtb脂质和蛋白质组合成单一亚单位疫苗制剂， 常规和非常规T细胞亚群可增强对Mtb感染的总体免疫力。的方法 本研究中开发的抗原/佐剂递送系统将指导下一代多亚单位 用于结核病和其他细菌病原体的疫苗，以提供快速疫苗制造的可扩展途径。

PROJECT SUMMARY Subunit vaccines combine immunodominant protein or peptide antigens from pathogens with select adjuvants, aiming to provide a more scalable, reproducible, low cost and rapid alternative to attenuated vaccines that contain live pathogens. Unfortunately, current subunit vaccines lack lipid antigens and rarely achieve the broad T cell responses required for lasting immunological memory and protection. In contrast, attenuated vaccines lack customization and scalability, but incorporate the entire pathogen to provide both protein and lipid antigens during immunization. This combination of lipid and protein antigens activates a broad spectrum of effector T cells, including conventional MHC-restricted T cells that respond to peptides and display considerable polymorphism, as well as nonpolymorphic CD1-restricted T cells that are directed against specific lipids. A more biomimetic strategy that simultaneously activates both lipid- and peptide-specific T cells may therefore show enhanced efficacy and control compared to subunit vaccines limited to protein antigens. The neglect of lipid antigens from current subunit vaccines and immunotherapies is primarily due to 1) difficulties in targeted delivery of lipids, and 2) a lack of suitable mouse models. In humans, the CD1 family consists of group 1 CD1 molecules (CD1a, CD1b, and CD1c) and the group 2 CD1 molecule CD1d. Mice, however, only express CD1d. This project, which involves a close collaboration between research groups led by a bioengineer and a basic immunologist, aims to overcome these obstacles by designing nanobiomaterials for enhanced dual delivery of both lipid and protein antigens in combination with adjuvants to induce CD1- and MHC- restricted T cell response in humanized CD1 transgenic (hCD1Tg) mice. To characterize, optimize and benchmark these novel nanobiomaterials against the most frequently used attenuated vaccine in the world, the bacillus Calmette-Guérin (BCG) tuberculosis (TB) vaccine, the following aims are proposed: In Aim 1, in vitro and in vivo approaches will identify the optimal nanobiomaterials and adjuvant combination for eliciting a combined CD1- and MHC-restricted T cell response. In Aim 2, a lipid/protein multi-antigen approach will be validated in hCD1Tg mice challenged with virulent Mycobacterium tuberculosis (Mtb). In Aim 3, a novel hydrogel delivery system will be employed for controlled and sustained release of lipid-antigen-loaded nanobiomaterials to assess efficacy and safety of chronic CD1-restricted T cell activation. The proposed study will provide a “proof of concept” that combining Mtb lipids and proteins into a single subunit vaccine formulation that targets both conventional and unconventional T cell subsets can enhance overall immunity to Mtb infection. The methodology and antigen/adjuvant delivery systems developed in this study will guide the next generation of multi-subunit vaccines for TB and other bacterial pathogens to provide scalable routes of rapid vaccine fabrication.