Immunological Secretomes of the Early Anthrax Infection

早期炭疽感染的免疫分泌组

• 批准号: 7439039
• 负责人: CHUN-MING HUANG
• 金额: $ 23.1万
• 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 2011-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7439039
• 关键词: Alveolar Macrophages; Anthrax Vaccines; Anthrax disease; Antigen Targeting; Antigens; Bacillus anthracis; Bacillus anthracis spore; Blood; Blood capillaries; Breathing; CAT development factor; Cells; Cessation of life; Data; Disease; Doctor of Philosophy; Drug Delivery Systems; Electrophoresis; Evaluation; Event; Germination; Growth; In Vitro; Infection; LIF gene; 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; cyclophosphamide/doxorubicin/fluorouracil protocol; cytotoxicity; edema factor; immunogenic; immunogenicity; in vivo; lymph nodes; macrophage; novel; novel vaccines; 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）：我们计划开发针对炭疽芽孢杆菌早期感染期间释放的蛋白质/肽（分泌组）的新型疫苗。在过去的几年里，我们已经建立了蛋白质组学技术来鉴定新的炭疽抗原。我们的实验室已经构建了几种编码这些新型炭疽抗原的载体疫苗。此外，我们通过鉴定与萌发和早期生长相关的孢子蛋白，发表了炭疽芽孢杆菌的蛋白质组。然而，炭疽芽孢杆菌释放的蛋白/肽（分泌组）不包括在我们和其他已发表的炭疽蛋白组中。在本研究中，我们将主要采用质谱技术对炭疽菌分泌组进行二维电泳鉴定。我们将利用毛细管超滤（CDF）探针收集炭疽感染小鼠的分泌组，以捕获体内低丰度和分泌蛋白/肽。CDF探针是我们实验室最近开发的一种用于收集小鼠体内低丰度和分泌性蛋白质的新技术。我们计划重点研究炭疽感染早期释放的蛋白质/肽。我们的假设是，针对炭疽感染早期阶段（上游事件）的疫苗将阻断下游事件，包括保护性抗原（PA）、致死因子（LF）和水肿因子（EF）的产生。为了构建更有效的疫苗，我们将确定炭疽菌分泌组的细胞毒性和免疫原性。同时，与目前基于PA/ lf的疫苗相比，我们将评估编码早期炭疽感染分泌组的疫苗的有效性。最后，我们发现了一种叫做camelysin样蛋白（CLP）的新毒素，它在炭疽生命周期的早期阶段从休眠孢子中释放出免疫原性。因此，该蛋白将作为阳性对照来评估在拟议的研究中发现的新抗原。