Antibodies to Burkholderia type III secretion system

伯克霍尔德杆菌 III 型分泌系统抗体

• 批准号: 6686599
• 负责人: DONALD I STEWART
• 金额: $ 24.67万
• 依托单位: CANGENE CORPORATION
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-09-15 至 2006-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6686599
• 关键词: Burkholderia; Escherichia coli; active immunization; antigen antibody reaction; bacterial genetics; bacterial proteins; biotechnology; bioterrorism /chemical warfare; cooperative study; disease /disorder model; drug screening /evaluation; genetic library; immunoglobulin G; immunologic substance development /preparation; informatics; laboratory mouse; monoclonal antibody; polymerase chain reaction; recombinant proteins; virulence

DESCRIPTION (provided by applicant): B. mallei and B. pseudomallei ("B. (pseudo)mallei"), two closely related Gram-negative bacteria, are serious potential bioterrorist agents listed on CDC/MMWR's category B list. These organisms cause life-threatening infections (Glanders' disease and Melioidosis, respectively), where antibiotic treatment is sometimes insufficient. The virulence factors of the two species are not well understood, and it is difficult to distinguish these species from each other and also from the closely related avirulent species B. thailandensis. B.(pseudo)mallei is also a civilian problem causing septicemia in Southeast Asia. Another Burkholderia species, B. cepacia, is a more common civilian problem that increases mortality and morbidity of cystic fibrosis patients. To better treat and diagnose B.(pseudo)mallei, we will develop monoclonal antibodies (mAbs) against potential virulence factors: components of the type III secretion system (TTSS). The presence of TTSS has been correlated to virulence of Burkholderia. Also, since several other invasive Gram-negative bacteria utilize TTSS to enter host cells, it is plausible that TTSS are critical for Burkholderia's virulence as well. TTSS components will be cloned and expressed in E.coli to generate sufficient quantities of purified antigen to generate mAbs. Fully human mAbs will be generated by phage display technology. Initial screening of the mAbs for reactivity will be conducted against a panel of Burkholderia isolates cultivated under different conditions, mAbs of interest will be further evaluated for protective capability in both in vitro and subsequently in vivo small animal models. We will also investigate potential differences of TTSS genes among isolates of Burkholderia. In summary the primary objectives of the project are: -Development of mAbs with utility in diagnosing and treating B.(pseudo)mallei and other Burkholderia species, including B.cepacia. -Improved understanding of the pathogenicity of B.(pseudo)mallei as a basis for potential vaccine development.

描述（由申请人提供）：两种密切相关的革兰氏阴性细菌是B. Mallei和B. Pseudomallei（“ B.（Pseudo）Mallei”），是CDC/MMWR类别B列表中列出的严重潜在的潜在生物恐怖剂。这些生物会引起威胁生命的感染（分别是腺体疾病和黑胶病），其中抗生素治疗有时不足。这两个物种的毒力因子尚不清楚，很难将这些物种彼此区分开，也很难与泰国人的密切相关的无毒物种区分开。 B.（伪）Mallei也是引起东南亚败血症的平民问题。伯克霍尔德（B. cepacia）的另一种伯克霍尔德（B. cepacia）是一个更常见的平民问题，可增加囊性纤维化患者的死亡率和发病率。 为了更好地治疗和诊断B。（伪）Mallei，我们将针对潜在的毒力因子开发单克隆抗体（MAB）：III型分泌系统（TTSS）的成分。 TTS的存在与Burkholderia的毒力有关。同样，由于其他几种侵入性革兰氏阴性细菌利用TTSS进入宿主细胞，因此TTSS对Burkholderia的毒力也至关重要，这是合理的。 TTSS组件将在大肠杆菌中克隆并表达，以产生足够数量的纯化抗原以产生mAb。噬菌体展示技术将产生完全的人物mAB。将对MAB进行反应性的初步筛选将针对在不同条件下培养的Burkholderia分离株进行的，并将进一步评估感兴趣的单元单元于体外和体内小动物模型中的保护能力。我们还将调查伯克霍尔德分离株之间TTSS基因的潜在差异。总之，该项目的主要目标是： - 在诊断和治疗B.（伪）Mallei和其他Burkholderia物种（包括B.Cepacia）中，具有实用性的mABS开发。 - 对B.（伪）MALLEI的致病性的理解是潜在疫苗发育的基础。