Novel multivalent vaccine for anthrax and botulinum

新型炭疽和肉毒杆菌多价疫苗

• 批准号: 8321943
• 负责人: Mansour M Zadeh
• 金额: $ 67.82万
• 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/8321943
• 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）：有效的疫苗增强抗体活性，延长T细胞寿命。利用表达炭疽保护性抗原（PA）融合蛋白的益生菌乳酸杆菌（L.）菌株正在开发新一代粘膜疫苗。这种由益生菌乳酸菌表达的口服疫苗不仅能激活树突状细胞（dc），还能通过噬菌体文库中提取的12-mer肽将靶向抗原（如炭疽PA）递送到粘膜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，使用高拷贝载体进行PA-DC融合表达，对炭疽杆菌的攻击具有强大的免疫保护作用。本研究的目的是测试这种由炭疽PA和肉毒梭菌神经毒素血清型A （BoNT/A）重链（Hc）组成的口服多价疫苗在吸收式炭疽和BoNT/A中毒中的疗效，并最终检验将炭疽PA- dcpep和/或BoNT/A Hc- dcpep融合物的c端整合到L. gasseri染色体中是否优于基于质粒的方法。适用于这项工作的三种新方法是：(a)利用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来表达多价免疫原性融合疫苗，避免了潜在的质粒不稳定性。我们的假设是，这种优化的口服多价疫苗将诱导小鼠对吸入性炭疽和BoNT/A的粘膜和全身免疫反应。具体目的是：(1)确定表达靶向炭疽PA-DCpep或BoNT/A Hc-DCpep融合疫苗的L. gasseri是否能增强对吸收式炭疽或BoNT/A中毒的保护性免疫；(2)确定PA-DCpep或BoNT/A Hc-DCpep融合疫苗在L. gasseri染色体位置表达时的体内保护效果；(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吸入性炭疽和BoNT/A攻击的效果。这项工作将通过新型佐剂来完成，这些佐剂提供疫苗的“靶向性”，评估疫苗对致命病原体的有效性，以及控制益生菌L. gasseri基因表达，这些基因表达可以很容易地口服，从而使“靶向”抗原自然递送到粘膜dc。