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

项目总结 亚单位疫苗将来自病原体的免疫优势蛋白或多肽抗原与选择 佐剂，旨在提供一种更具可扩展性、可重复性、低成本和快速的减毒疫苗替代品 含有活的病原体。不幸的是，目前的亚单位疫苗缺乏脂类抗原，很少能达到 持久的免疫记忆和保护需要广泛的T细胞反应。相比之下，衰减 疫苗缺乏定制化和可扩展性，但结合了整个病原体来提供蛋白质和脂肪 免疫过程中的抗原。这种脂类和蛋白质抗原的组合激活了广泛的 效应性T细胞，包括传统的MHC限制性T细胞，对多肽有反应并表现出相当大的 多态，以及针对特定脂质的非多态CD1限制性T细胞。A更多 因此，同时激活脂质和多肽特异性T细胞的仿生策略可能会显示 与仅限于蛋白质抗原的亚单位疫苗相比，提高了效力和控制力。 目前的亚单位疫苗和免疫疗法忽略了脂类抗原，这主要是由于1) 2)缺乏合适的小鼠模型。在人类中，CD1家族 由第1组CD1分子(CD1a、CD1b和CD1c)和第2组CD1分子CD1d组成。老鼠， 然而，仅表达CD1d。该项目涉及由以下人员领导的研究小组之间的密切合作 一位生物工程师和一位基础免疫学家，旨在通过设计纳米生物材料来克服这些障碍 结合佐剂增强脂类和蛋白质抗原的双重递送以诱导CD1-和 人源化CD1转基因(HCD1Tg)小鼠MHC限制性T细胞反应要表征、优化和 将这些新型纳米生物材料与世界上使用最频繁的减毒疫苗 卡介苗(BCG)结核病疫苗，提出了以下目标：目标1，体外 体内方法将确定最佳的纳米生物材料和佐剂组合，以诱导 CD1和MHC联合限制的T细胞反应。在目标2中，脂质/蛋白质多抗原方法将是 在用强毒结核分枝杆菌(Mtb)攻击的hCD1Tg小鼠中得到验证。在《目标3》中，一种新型水凝胶 脂类抗原纳米生物材料的控释和缓释将采用递送系统 目的：评价慢性CD1限制性T细胞活化的有效性和安全性。拟议的研究将提供一个“证据” 将结核分枝杆菌脂类和蛋白质结合到一个单一的亚单位疫苗配方中，以此为目标 常规和非常规T细胞亚群可增强对结核分枝杆菌感染的整体免疫力。方法论 本研究开发的抗原/佐剂递送系统将指导下一代多亚单位 针对结核病和其他细菌病原体的疫苗，提供可扩展的快速疫苗制造路线。

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.