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，主要需要构建安全的分子佐剂，包括那些旨在保护免受A类病原体和外毒素影响的疫苗。在这项赠款之前的工作集中在通过使用经典的粘膜佐剂霍乱毒素(CT)的新的无毒衍生物来确定小鼠鼻部相关淋巴网状组织(NALT)为诱导部位。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。这些研究还包括对人类NALT的特征，例如扁桃体和腺样体对艾滋病毒感染的特征，以及防止艾滋病毒性传播的粘膜和系统免疫的最佳发展。总共成功地实现了五个最初的具体目标。2001年9月11日事件和涉及炭疽污染信件的后果导致了评估粘膜对炭疽芽孢杆菌及其三种外毒素的免疫力的重要新举措。为此，我们选择利用CT的ADP-核糖化缺陷、CT的分子突变体和由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基于NALT的小鼠对炭疽的黏膜免疫。在这项续期拨款中，我们提出了五个具体的目标，这些目标完成后，将提供重要的新信息，即在体外和体内诱导粘膜分泌型免疫球蛋白A(S-免疫球蛋白A)和血清免疫球蛋白亚类和免疫球蛋白A抗体(抗体)，以及它们在预防炭疽病中的作用。第一个特定目的是比较无毒的MCT和给予保护性抗原(PA)的天然CT诱导保护性S-Ig A抗体的效果。第二个具体目标是鉴定一组新的单抗，其中包括所有四个免疫球蛋白亚类，即IgE和IgA。第三个具体目标是利用新开发的短杆菌表达系统生产MCT和MCT-A/LT-B嵌合体佐剂和PA融合蛋白，作为潜在的鼻腔疫苗来预防吸入性炭疽。第四个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S-IgA抗PA单抗中和炭疽外毒素的体外研究。最后一个具体目标是建立炭疽组分鼻腔给药的体内模型，以开发一种评估鼻腔炭疽毒性和粘膜免疫的方法。