MOLECULAR ADJUVANTS FOR NALT-BASED IMMUNITY TO ANTHRAX

基于 NALT 的炭疽免疫分子佐剂

• 批准号: 6615012
• 负责人: Prosper N Boyaka
• 金额: $ 36.25万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-03-15 至 2008-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6615012
• 关键词: Bacillus anthracis; anthrax; anthrax vaccines; antibody neutralization test; bacterial toxins; bactericidal immunity; bioterrorism /chemical warfare; chimeric proteins; cholera toxin; enzyme linked immunosorbent assay; exotoxins; flow cytometry; immunoglobulin A; immunoglobulin E; immunoglobulin G; immunomodulators; inhalation drug administration; laboratory mouse; laboratory rat; monoclonal antibody; mucosal immunity; neutralizing antibody; respiratory epithelium; vaccine development

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.

描述（由申请人提供）：在新的粘膜疫苗开发中构建安全分子佐剂的主要需求，包括旨在保护A.病原体和外毒素的那些。这项赠款的先前工作重点是将鼠鼻鼻相关组织（NALT）定义为电感部位，通过使用新型的经典粘膜辅助辅助霍乱毒素（CT）的无毒衍生物。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。这些研究还包括对HIV感染的扁桃体和腺样体的特征，以及粘膜和系统免疫的最佳发展，以防止HIV的性传播。总共成功解决了五个原始特定目标。 2001年9月11日的事件以及涉及炭疽染色字母的后果导致了重要的新举措，以评估对炭疽芽孢杆菌及其三个部分Exotoxin的粘膜免疫。因此，我们选择使用由CT和CT和B亚基组成的CT和嵌合体的ADP-核糖化缺陷，CT和嵌合体的分子突变体研究基于鼠NALT的粘膜免疫。在这项续签赠款中，我们提出了五个具体目标，完成后将提供有关粘膜分泌性IgA（S-IgA）和血清IgG亚类和IgA抗体（ABS）的基本新信息，以防止炭疽病，包括体外和体内。第一个特定目的将将无毒的MCT与给出的天然CT与保护性抗原（PA）进行比较，以诱导保护性S-IGA ABS。第二个特定目标将表征新的单克隆抗PA ABS（mAb），其中包括所有四个IgG亚类，IgE和IgA。第三个特定目标将采用新开发的Brevibacillus表达系统来生产MCT和MCT-A / LT-B嵌合体辅助剂和PA融合蛋白作为潜在的鼻疫苗，以保护免受吸入炭疽。第四个特定目的将建立体外呼吸道上皮细胞，对炭疽的外毒素的敏感性，用于对S-IGA抗PA MAB的体外研究，以中和炭疽异毒素。最后的特定目的将建立一种鼻施用炭疽成分的体内模型，以开发一种评估鼻炭疽毒性和粘膜免疫的方法。