Immunological Secretomes of the Early Anthrax Infection

早期炭疽感染的免疫分泌组

基本信息

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
项目类别：
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

项目摘要

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（申请人提供）：我们计划开发针对炭疽杆菌早期感染过程中释放的蛋白质/肽（分泌组）的新型疫苗。在过去的几年中，我们已经建立了蛋白质组学技术来鉴定新型炭疽抗原。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载体的疫苗。此外，我们通过鉴定与发芽和早期生长相关的孢子蛋白，发表了炭疽杆菌的蛋白质组。然而，炭疽杆菌释放的蛋白质/肽（分泌组）不包含在我们和其他已发表的炭疽蛋白质组中。在本提案中，我们将主要采用质谱技术来鉴定炭疽分泌物，通过/不通过二维电泳进行蛋白质分离。我们将使用毛细管超滤（CDF）探针从炭疽感染的小鼠中收集分泌蛋白组，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项新技术，用于从小鼠中收集低丰度和分泌蛋白。我们计划重点关注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，我们将确定炭疽分泌物的细胞毒性和免疫原性。与此同时，我们将评估编码早期炭疽感染分泌体的疫苗与目前基于 PA/LF 的疫苗的功效。最后，我们发现了一种称为骆驼素样蛋白（CLP）的新型毒素，它具有免疫原性，并在炭疽生命周期的早期阶段从休眠孢子中释放出来。因此，该蛋白质将作为阳性对照来评估拟议研究中鉴定的新抗原。