Novel multivalent vaccine for anthrax and botulinum

新型炭疽和肉毒杆菌多价疫苗

• 批准号: 8707184
• 负责人: Mansour M Zadeh
• 金额: $ 57.81万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-08-19 至 2016-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8707184
• 关键词: Adjuvant; Adjuvanticity; Adverse effects; Affect; Aluminum Hydroxide; Animals; Anthrax Vaccines; Anthrax disease; Antibiotic Resistance; Antibiotics; Antibody Avidity; Antigen Targeting; Antigens; Applications Grants; Bacillus anthracis; Bacteria; Bacteriophages; Binding; Bontoxilysin; Botulism; Breathing; C-terminal; Catalytic Domain; Cell fusion; Cell physiology; Cells; Cessation of life; Chimeric Proteins; Chromosomal Insertion; Chromosomes; Clostridium botulinum; Cytosol; Data; Dendritic Cells; Dendritic cell activation; Dose; Endosomes; Enzymes; Gastrointestinal tract structure; Gene Expression; Generations; Genetic; Genetic Recombination; Genome; Goals; Homo; Human; Immune; Immune response; Immune system; Immunity; Individual; Infection; Intoxication; Lactobacillus; Lactobacillus acidophilus; Libraries; Light; Link; Location; Longevity; Membrane; Mus; N-terminal; Neurons; Oral; Pathogenesis; Peptide Phage Display Library; Peptides; Plasmids; Probiotics; Production; Receptor Cell; Serotyping; Subcutaneous Injections; Subunit Vaccines; System; Systemic infection; T cell response; T-Lymphocyte; Testing; Toxin; Vaccines; Work; anthrax protective factor; antigen binding; base; botulinum; combat; cost effective; dietary supplements; disulfide bond; efficacy testing; expression vector; immunogenic; improved; in vivo; microbial; mortality; mucosal vaccine; novel; novel strategies; oral vaccine; pathogen; promoter; protective efficacy; receptor binding; response; targeted delivery; tool; vaccine delivery; vaccine effectiveness; vaccine evaluation; vector

DESCRIPTION (provided by applicant): Effective vaccines potentiate antibody avidity and increase T cell longevity. A new generation of mucosal vaccines is being developed using probiotic Lactobacillus (L.) strains expressing anthrax-protective antigen (PA) fusion protein. Such an oral vaccine, expressed by probiotic Lactobacillus species not only optimally activates dendritic cells (DCs), but also delivers targeted antigens (i.e. anthrax PA) to mucosal DCs via 12-mer peptides derived from a bacteriophage library. We further optimized the expression of the immunogenic vaccine fusions expressed by L. gasseri. Obtained data clearly show that the use of a high copy vector for PA-DC fusion expression by L. gasseri confers robust immune protection against anthrax Sterne challenge. The objective of this proposal is to test the efficacy of such an oral multivalent vaccine consisting of anthrax PA and Clostridium botulinum neurotoxin serotype A (BoNT/A) heavy chain (Hc) in inhaled anthrax and BoNT/A intoxication and lastly to examine whether integration of the anthrax PA-DCpep and/or BoNT/A C-terminus of Hc-DCpep fusions into the L. gasseri chromosome is superior to plasmid-based approaches. Three novel approaches adapted for this work are: (a) improving adjuvanticity of the delivery vector by employing L. gasseri; (b) improving vaccine potency by specifically using a high copy vector for immunogenic fusions; and (c) expressing the multivalent immunogenic fusion vaccines in L. gasseri by bacterial chromosomal insertion and avoiding potential plasmid instability. Our hypothesis is that this optimized oral multivalent vaccine will induce mucosal and systemic immune responses against inhaled anthrax and BoNT/A in mice. The specific aims are: (1) to determine whether L. gasseri expressing targeted anthrax PA-DCpep or BoNT/A Hc-DCpep fusion vaccines enhances protective immunity against inhaled anthrax or BoNT/A intoxication; (2) to determine the protective efficacy of PA-DCpep or BoNT/A Hc-DCpep fusion vaccines in vivo, when expressed from a chromosomal location in L. gasseri; and (3) to determine the efficacy of the probiotic multivalent vaccine consisting of anthrax-PA and BoNT/A-Hc fusions against inhalational anthrax and BoNT/A challenge. This effort will be accomplished via novel adjuvants that provide "targeting" of the vaccine, evaluation of vaccine effectiveness against deadly pathogens, and the control of probiotic L. gasseri gene expression that can readily be orally consumed to enable natural delivery of "targeted" antigen to mucosal DCs. Activation of a group of cells in our immune system, called dendritic cells, is critical for proper defense against different types of infection. The goal of this grant proposal is to provide tools for efficient activation of mucosal and systemic immune system using a new generation of harmless immunostimulatory bacteria called Lactobacillus gasseri that can be ingested as dietary supplements. Our data show that L. gasseri producing anthrax vaccine optimally generated robust protective immunity against anthrax infection. Therefore, this probiotic vaccine strategy will be used to show that they are important for multivalent vaccine delivery as well as critical vehicles that stimulate mucosal and systemic immunity for combating deadly bacteria such as anthrax and botulinum.

描述（由申请人提供）：有效的疫苗可增强抗体亲合力并延长 T 细胞寿命。新一代粘膜疫苗正在使用表达炭疽保护性抗原 (PA) 融合蛋白的益生菌乳杆菌 (L.) 菌株进行开发。这种由益生菌乳杆菌表达的口服疫苗不仅可以最佳地激活树突状细胞 (DC)，而且还可以通过源自噬菌体库的 12 聚体肽将目标抗原（即炭疽 PA）递送至粘膜 DC。我们进一步优化了加氏乳杆菌表达的免疫原性疫苗融合体的表达。获得的数据清楚地表明，L.gasseri 使用高拷贝载体进行 PA-DC 融合表达可提供针对炭疽 Sterne 攻击的强大免疫保护。本提案的目的是测试由炭疽 PA 和肉毒杆菌神经毒素 A 血清型 (BoNT/A) 重链 (Hc) 组成的口服多价疫苗在吸入炭疽和 BoNT/A 中毒中的功效，最后检查整合是否有效炭疽 PA-DCpep 和/或 Hc-DCpep 融合的 BoNT/A C 末端到 L.gasseri 染色体中的优于基于质粒的方法。适用于这项工作的三种新颖方法是：（a）通过采用 L.gasseri 提高递送载体的佐剂性； (b) 通过专门使用高拷贝载体进行免疫原性融合来提高疫苗效力； (c)通过细菌染色体插入在L.gasseri中表达多价免疫原性融合疫苗并避免潜在的质粒不稳定。我们的假设是，这种优化的口服多价疫苗将诱导小鼠针对吸入炭疽和 BoNT/A 的粘膜和全身免疫反应。具体目的是：（1）确定L.gasseri表达靶向炭疽PA-DCpep或BoNT/A Hc-DCpep融合疫苗是否增强针对吸入性炭疽或BoNT/A中毒的保护性免疫力； (2) 确定 PA-DCpep 或 BoNT/A Hc-DCpep 融合疫苗在 L.gasseri 中的染色体位置表达时的体内保护功效； (3)确定由炭疽-PA和BoNT/A-Hc融合物组成的益生菌多价疫苗对抗吸入性炭疽和BoNT/A攻击的功效。这项工作将通过新型佐剂来实现，这些佐剂提供疫苗的“靶向”、评估疫苗针对致命病原体的有效性，以及控制益生菌 L.gasseri 基因表达，这些益生菌可以轻松口服，从而能够自然递送“靶向”抗原至粘膜 DC。 激活我们免疫系统中的一组细胞（称为树突状细胞）对于正确防御不同类型的感染至关重要。该拨款提案的目标是提供使用新一代无害免疫刺激细菌（称为加氏乳杆菌）有效激活粘膜和全身免疫系统的工具，该细菌可以作为膳食补充剂摄入。我们的数据表明，生产炭疽疫苗的加氏乳杆菌能够最佳地产生针对炭疽感染的强大保护性免疫力。因此，这种益生菌疫苗策略将用于证明它们对于多价疫苗递送以及刺激粘膜和全身免疫以对抗炭疽和肉毒杆菌等致命细菌的关键载体非常重要。