Immunological Secretomes of the Early Anthrax Infection

早期炭疽感染的免疫分泌组

• 批准号: 7647403
• 负责人: CHUN-MING HUANG
• 金额: $ 20.6万
• 依托单位: VETERANS MEDICAL RESEARCH FDN/SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-03-01 至 2010-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7647403
• 关键词: Alveolar Macrophages; Anthrax Vaccines; Anthrax disease; Antigen Targeting; Antigens; Bacillus anthracis; Bacillus anthracis spore; Blood; Blood capillaries; Breathing; Cells; Cessation of life; Data; Disease; Doctor of Philosophy; Drug Delivery Systems; Electrophoresis; Evaluation; Event; Germination; Growth; In Vitro; Infection; Laboratories; Lead; Life; Life Cycle Stages; Methods; Mus; PAWR protein; Peptides; Production; Protein Databases; Proteins; Proteome; Proteomics; Publishing; Reproduction spores; Route; Site; Staging; Surface; Symptoms; Techniques; Toxin; Travel; Ultrafiltration; Vaccination; Vaccines; anthrax lethal factor; anthrax toxin; base; bioweaponry; camelysin; capillary; cytotoxicity; edema factor; immunogenic; immunogenicity; in vivo; lymph nodes; macrophage; novel; novel vaccines; protein profiling; secretory protein; success; vaccine efficacy; vaccine evaluation; vector; vector-based vaccine

DESCRIPTION (provided by applicant): We plan to develop the novel vaccines targeting the released proteins/peptides (secretome) during the early infection of Bacillus anthracis. In the past years, we had established the proteomic techniques to identify novel anthrax antigens. Several vector-based vaccines encoding these novel anthrax antigens have been constructed in our laboratory. In addition, we have published a proteome of Bacillus anthracis by identifying spore proteins associated with germination and early outgrowth. However, the released proteins/peptides (secretome) of Bacillus anthracis are not included in our and other published anthrax proteomes. In this proposal, we will mainly employ the mass spectrometric techniques to identify the anthrax secretomes with/without protein separation by 2-D electrophoresis. We will collect the secretomes from anthrax-infected mice using capillary ultrafiltration (CDF) probes in order to capture the in vivo low abundant and secretory proteins/peptides. The CDF probes are a novel technique recently developed in our laboratory to collect low abundant and secretory proteins from mice. We plan to emphasize on the proteins/peptides released during the early stages of anthrax infection. Our hypothesis is that vaccines targeting the early stages of anthrax infection (upstream events) will block the downstream events including the production of protective antigen (PA), lethal factor (LF), and edema factor (EF). In order to construct more effective vaccines, we will determine the cytotoxicities and immunogenicities of anthrax secretomes. In parallel, we will evaluate the efficacies of vaccines which encoded secretomes of early anthrax infection as compared to current PA/LF-based vaccines. Lastly, we have identified a novel toxin called camelysin-like protein (CLP) which exerts immunogenic and is released from dormant spores during the early stage of the anthrax life cycle. This protein will thus serve as a positive control to evaluate novel antigens identified in the proposed studies.

描述（由申请人提供）：我们计划在早期感染炭疽芽孢杆菌期间开发针对释放的蛋白/肽（分泌组）的新型疫苗。在过去的几年中，我们建立了蛋白质组学技术来识别新型炭疽抗原。我们的实验室已经构建了几种编码这些新型炭疽抗原的基于向量的疫苗。此外，我们通过鉴定与发芽和早期生长相关的孢子蛋白来发表炭疽芽孢杆菌的蛋白质组。然而，我们和其他已发表的炭疽蛋白质组中未包含释放的炭疽芽孢杆菌蛋白/肽（分泌组）。在此提案中，我们将主要采用质谱技术来识别2-D电泳分离/没有蛋白质分离的炭疽秘密组。我们将使用毛细管超滤清（CDF）探针从炭疽感染的小鼠那里收集秘密组，以捕获体内低的丰富和分泌蛋白/肽。 CDF探针是一种在我们的实验室中开发的一种新型技术，可从小鼠收集低丰富和分泌的蛋白质。我们计划强调在炭疽感染的早期阶段释放的蛋白质/肽。我们的假设是，针对炭疽感染早期（上游事件）的疫苗将阻止下游事件，包括生产保护性抗原（PA），致命因子（LF）和水肿因子（EF）。为了构建更有效的疫苗，我们将确定炭疽秘密组的细胞毒性和免疫原性。同时，我们将评估与当前基于PA/LF的疫苗相比，它们编码早期炭疽感染的疫苗的效力。最后，我们确定了一种称为Camelysin样蛋白（CLP）的新型毒素，该毒素在炭疽藻生命周期的早期阶段发挥免疫原性并从休眠孢子中释放出来。因此，该蛋白将作为评估拟议研究中鉴定出的新型抗原的阳性对照。