Alternative approaches for NALT-based immunity to respiratory pathogens

摘要 鼻咽道是致衰性和潜在致命性病原体的主要入口。 包括炭疽杆菌和流感病毒。能促进抗体和免疫的黏膜疫苗 粘膜组织中的细胞毒性T细胞反应，除了一般的血流外，还能保护更多 比传统的注射疫苗更有效地对抗呼吸道病原体。因此，有必要 用于安全的粘膜佐剂和疫苗输送系统。之前关于这笔赠款的工作主要集中在 确定小鼠鼻相关淋巴网状组织(NALT)为免疫诱导部位 以鼻咽部为目标的反应。我们使用了霍乱毒素并开发了新的衍生品 缺乏ADP核糖基转移酶活性以绕过这种肠毒素佐剂的反应性。 此外，我们还评估了炭疽毒素对小鼠NALT的作用。这些研究还 包括炭疽保护性抗原抗体(Abs)的特性，特别强调 关于保护粘膜组织的潜力。总共成功实现了五个原创的具体目标 地址。在我们的研究过程中，一个重要的和意想不到的发现是，鼻部 炭疽杆菌保护性抗原与cAMP诱导型突变体联合应用的研究 炭疽芽孢杆菌水肿素增强对这两种细菌的粘膜和系统免疫 分子。此外，我们还发现，水肿性毒素(EdTx或保护性抗原+水肿性 因子)衍生物增强联合注射无关抗原的黏膜和系统免疫 如重组鼠疫F1-V抗原。与神经节苷脂结合的霍乱肠毒素不同 毒素，EdTx及其衍生物在鼻腔应用后不会针对中枢神经系统组织。在……里面 在这次续订拨款中，我们将解决EdTx的舌下应用的总体假设 衍生物将诱导基于NALT的免疫并保护呼吸道病原体，而不是 不良反应通常与鼻腔应用肠毒素有关。一个人将确立的具体目标 与保护性抗原(PA)联合给药的水肿素衍生物的佐剂活性 增强对炭疽毒素成分的免疫力的舌下途径。具体目标二将 舌下免疫对呼吸道病毒感染的保护性免疫特征 EdTx衍生物作为佐剂。在特定目标三的研究将确定诱导位置 舌下免疫后分泌型IgA抗体的产生及粘膜免疫 EdTx-衍生品。最后，特定的目标四将确定潜在的分子信号诱导 作为舌下佐剂的EdTx-衍生物对SIgA的应答。这笔赠款将解开这些机制 通过EdTx衍生物作为舌下疫苗的佐剂，诱导基于NALT的免疫。 我们还将验证一种新的粘膜疫苗递送途径，以及基于PA的新粘膜疫苗 佐剂(S)，用于诱导对呼吸道病原体的免疫。

Abstract The nasopharyngeal tract is a major portal entry of debilitating and potentially lethal pathogens including Bacillus anthracis and influenza virus. Mucosal vaccines capable of promoting antibody and cytotoxic T cell responses in mucosal tissues, in addition to the general bloodstream, protect more effectively against respiratory pathogens than classical injected vaccines. Therefore, there is a need for safe mucosal adjuvants and vaccine delivery systems. Previous work on this grant focused at defining murine nasal-associated lymphoreticular tissues (NALT) as inductive sites for immune responses targeting the nasopharyngeal tract. We used cholera toxin and developed novel derivatives lacking ADP ribosyl transferase activity to circumvent the reactogenicity of this enterotoxin adjuvant. In addition, we have assessed the action of anthrax toxins on murine NALT. These studies have also included the characterization of antibodies (Abs) to anthrax protective antigens with special emphasis on the potential to protect mucosal tissues. A total of five original Specific Aims were successfully addressed. A major and unexpected finding during the course of our studies was the fact the nasal co-administration of Bacillus anthracis protective antigen together with a mutant of the cAMP-inducing Bacillus anthracis edema factor enhanced mucosal and systemic immunity against these two molecules. Furthermore, we found that the edema toxin (EdTx or protective antigen plus edema factor) derivative enhanced mucosal and systemic immunity to co-administered unrelated antigens such as recombinant Yersinia pestis F1-V antigen. Unlike the ganglioside-binding enterotoxin cholera toxin, neither EdTx nor its derivatives target central nervous system tissues after nasal application. In this renewal grant, we will address the overall hypothesis that sublingual application of EdTx derivatives will induce NALT-based immunity and protect against respiratory pathogens, without the adverse effects often associated with nasal application of enterotoxins. Specific aim one will establish the adjuvant activity of edema factor derivatives co-administered with protective antigen (PA) via the sublingual route for enhanced immunity to anthrax toxin components. Specific aim two will characterize protective immunity to respiratory viral infection afforded by sublingual immunization with EdTx derivatives as adjuvant. Studies in specific aim three will identify inductive sites for the generation of secretory IgA (SIgA) Abs and mucosal immunity after sublingual immunization with EdTx-derivatives. Finally, specific aim four will determine molecular signals underlying the induction of SIgA responses by EdTx-derivatives as sublingual adjuvant. This grant will unravel the mechanisms by which EdTx derivatives act as adjuvant for sublingual vaccines and induce NALT-based immunity. We will also validate a new route for mucosal vaccine delivery, as well as new PA-based mucosal adjuvant(s) for the induction of immunity against respiratory pathogens.