The analysis of the disease-causing gene of the tubercle bacillus and fundamental examination of tuberculosis attack control of the candidate new vaccine strain

候选新疫苗株结核杆菌致病基因分析及控制结核病发作的基础检验

• 批准号: 17590794
• 负责人: MIYAZAKI Yoshitsugu
• 金额: $ 1.22万
• 依托单位: Nagasaki University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2005
• 资助国家: 日本
• 起止时间: 2005 至 2006
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-17590794/
• 关键词: tuberculosis; latent infection; macrophage; NO; RNI; ctpF

The tuberculosis attracts attention as reemerging infectious disease in late year. 60% of the tuberculosis patient are senior citizens in Japan. The most them are past infection patients. and it is thought as follows, and, as for becoming sick through a life among tuberculosis infection patients, it is thought that a tuberculosis patient equal to or less than 10% has tuberculosis for a life. The macrophage is important as host defense mechanism of the tuberculosis infection. There is NO (nitric oxide) and RNI (reactive nitrogen intermediates) in one of the defense mechanism of a host against tuberculosis. We got the research finding that a ctp F gene may participate in a tubercle bacillus greatly as genetic mechanism of the escape mechanism from this defense mechanism. Our object of this research is the confirmation of this result and elucidation of a concrete function of these genes. We knocked out of ctp F gene by allelic exchange method to make gene deficit strain. We examine influence of the knock out strain revealed by NO and RNI in vitro and compare it with a wild strains. In addition, we let these knock outs strain infect with human THP-1 macrophages and compare knock out strain with wild strains and examine the change of the number of viable bacteria, the viable or death of the cell, and an apoptosis instruction over time. In the future, we perform in vivo experiments with mice.

结核病作为一种晚年重发传染病而备受关注。在日本，60%的肺结核患者是老年人。其中大多数是过去的感染患者。人们是这样认为的，对于肺结核感染患者一生患病的情况，人们认为肺结核患者一生中患病的人数等于或少于10%巨噬细胞在结核感染的宿主防御机制中起着重要作用。在宿主抗结核的防御机制中存在NO（一氧化氮）和RNI（活性氮中间体）。我们得到的研究发现，一个ctp F基因可能在很大程度上参与了结核杆菌从这种防御机制中逃脱的遗传机制。我们这项研究的目的是确认这一结果，并阐明这些基因的具体功能。我们用等位基因交换法敲除ctp - F基因，制成基因缺失菌株。我们考察了NO和RNI对体外敲除菌株的影响，并与野生菌株进行了比较。此外，我们将这些敲除菌株与人THP-1巨噬细胞感染，并将敲除菌株与野生菌株进行比较，观察随时间的变化，活菌数量，细胞存活或死亡，以及凋亡指令。在未来，我们将在小鼠身上进行体内实验。