MOLECULAR ADJUVANTS FOR NALT-BASED IMMUNITY TO ANTHRAX

描述（由申请方提供）：在新的粘膜疫苗开发中，主要需要构建安全的分子佐剂，包括设计用于保护A类疫苗的分子佐剂。病原体和外毒素。这项资助的前期工作主要集中在使用经典粘膜佐剂霍乱毒素（CT）的新型无毒衍生物将小鼠鼻相关淋巴网状组织（NALT）定义为诱导位点。此外，我们在这些研究中评估了调节性细胞因子和趋化因子，以表征基于NALT的粘膜免疫。这些研究还包括人NALT的表征，例如，扁桃体和腺样体，用于HIV感染以及粘膜和全身免疫的最佳发展，以防止HIV的性传播。共有五个最初的具体目标得到成功解决。2001年9月11日的事件和炭疽污染信件的后果导致了重要的新举措，以评估粘膜免疫炭疽杆菌及其三部分外毒素。为此，我们选择使用ADP核糖基化缺陷的CT分子突变体和由CT的A亚基和大肠杆菌不稳定毒素（LT）的B亚基组成的嵌合体来研究小鼠对炭疽的基于NALT的粘膜免疫。在这次更新补助金中，我们提出了五个具体目标，完成后，将提供有关粘膜分泌型伊加（S-IgA）和血清IgG亚类和伊加抗体（Abs）在体外和体内保护炭疽中的诱导和功能的重要新信息。第一个特定目的将比较无毒mCT与给予保护性抗原（PA）的天然CT，以诱导保护性S-IgA Ab。第二个特异性目的将表征一组新的单克隆抗PA抗体（mAb），包括所有四个IgG亚类，IgE和伊加。第三个Specific Aim将采用新开发的短芽孢杆菌表达系统生产mCT和mCT-A / LT-B嵌合佐剂和PA融合蛋白，作为潜在的鼻疫苗，以防止吸入性炭疽。第四个具体目标是建立对炭疽外毒素敏感的体外呼吸道上皮细胞，用于中和炭疽外毒素的S-IgA抗PA mAb的体外研究。最后一个具体目标将建立一个体内模型，鼻炭疽成分的管理，以开发一种方法来评估鼻炭疽毒性和粘膜免疫。

DESCRIPTION (provided by applicant): There is a major need to construct safe molecular adjuvants in new mucosal vaccine development, including those designed to protect from Category A. pathogens and exotoxins. Previous work on this grant focused on defining murine nasal-associated lymphoreticular tissues (NALT) as inductive sites by use of novel nontoxic derivatives of the classical mucosal adjuvant cholera toxin (CT). In addition, we have assessed regulatory cytokines and chemokines in these studies to characterize NALT-based mucosal immunity. These studies have also included characterization of human NALT, e.g., the tonsils and adenoids for both HIV infection as well as for optimal development of mucosal and systemic immunity to prevent sexual transmission of HIV. A total of five original Specific Aims were successfully addressed. The events of September 11, 2001 and the aftermath involving anthrax-tainted letters has led to important new initiatives to assess mucosal immunity to Bacillus anthracis and its three part exotoxin. For this reason, we have chosen to study murine NALT-based mucosal immunity to anthrax using the ADP-ribosylation deficient, molecular mutants of CT and chimeras consisting of A subunit of CT and B subunits of Escherichia coil labile toxin (LT). In this renewal grant, we have proposed five Specific Aims, which when completed, will provide essential new information regarding induction of and the functions for mucosal secretory IgA (S-IgA) and serum IgG subclass and IgA antibodies (Abs) in protection from anthrax, both in vitro and in vivo. The first Specific Aim will compare nontoxic mCTs with native CT given with protective antigen (PA) for induction of protective S-IgA Abs. The second Specific Aim will characterize a new panel of monoclonal anti-PA Abs (mAbs) which include all four IgG subclasses, IgE and IgA. The third Specific Aim will employ a newly developed Brevibacillus expression system to produce mCTs and mCT-A / LT-B chimera adjuvants and PA fusion protein as a potential nasal vaccine to protect from inhalational anthrax. The fourth Specific Aim will establish in vitro respiratory epithelial cells with sensitivity to the exotoxin of anthrax for in vitro studies of S-IgA anti-PA mAbs for neutralization of anthrax exotoxin. The last Specific Aim will establish an in vivo model of nasal administration of anthrax components for development of a method to assess nasal anthrax toxicity and mucosal immunity.