MOLECULAR ADJUVANTS FOR NALT-BASED IMMUNITY TO ANTHRAX

基于 NALT 的炭疽免疫分子佐剂

• 批准号: 7197985
• 负责人: Prosper N Boyaka
• 金额: $ 34.15万
• 依托单位: OHIO STATE UNIVERSITY
• 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 2008-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7197985
• 关键词: ADP Ribose Transferases; ADP ribosylation; Address; Adenoidal structure; Adjuvant; Adjuvanticity; Amino Acid Substitution; Anthrax disease; Antibodies; Antigens; Bacillus anthracis; Categories; Chimera organism; Chimeric Proteins; Cholera Toxin; Cholera Toxin Protomer B; Development; Edema; Epithelial Cells; Escherichia; Event; Exotoxins; Goals; Grant; HIV; HIV Infections; Human; Hybrids; IgE; Immune response; Immunity; Immunization; Immunoglobulin A; Immunoglobulin G; In Vitro; Letters; Mediating; Methods; Modeling; Molecular; Mucosal Immunity; Mus; Nose; Reproduction spores; Secretory Immunoglobulin A; Serum; Sexual Transmission; Site; System; Tissues; Tonsil; Toxic effect; Toxin; Vaccine Antigen; Vaccines; Work; anthrax lethal factor; anthrax protective factor; anti-IgA; base; chemokine; cytokine; design; in vivo; in vivo Model; mucosal vaccine; mutant; novel; pathogen; prevent; respiratory; response; 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迫切需要构建安全的分子佐剂，包括那些旨在预防 A 类病原体和外毒素的佐剂。这项资助的先前工作重点是通过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。这些研究还包括人类 NALT 的特征，例如扁桃体和腺样体对于 HIV 感染以及粘膜和全身免疫的最佳发展以防止 HIV 性传播。总共五个最初的具体目标已成功实现。 2001 年 9 月 11 日的事件以及涉及炭疽污染信件的后果导致了评估对炭疽杆菌及其三部分外毒素的粘膜免疫的重要新举措。因此，我们选择使用 CT 的 ADP-核糖基化缺陷型分子突变体和由 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基于 NALT 的粘膜免疫。在这笔更新拨款中，我们提出了五个具体目标，这些目标完成后，将提供有关粘膜分泌型 IgA (S-IgA) 和血清 IgG 亚类和 IgA 抗体 (Abs) 的诱导和功能的重要新信息，以预防炭疽病，体外和体内。第一个具体目标将比较无毒 mCT 与给予保护性抗原 (PA) 的天然 CT，以诱导保护性 S-IgA 抗体。第二个具体目标将描述一组新的单克隆抗 PA 抗体 (mAb)，其中包括所有四种 IgG 亚类、IgE 和 IgA。第三个具体目标将采用新开发的短芽孢杆菌表达系统来生产 mCT 和 mCT-A / LT-B 嵌合佐剂以及 PA 融合蛋白，作为潜在的鼻疫苗来预防吸入性炭疽。第四个具体目标将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 单克隆抗体的体外研究。最后一个具体目标将建立炭疽成分鼻腔给药的体内模型，以开发评估鼻炭疽毒性和粘膜免疫的方法。