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

结核病作为近年来再次出现的传染病引起了人们的关注。在日本，60%的结核病患者是老年人。他们中的大多数是过去的感染患者。它被认为是这样的，至于在结核病感染患者中通过生命患病，等于或低于10%的结核病患者被认为是一生患有结核病。巨噬细胞是结核病感染的重要宿主防御机制。在宿主对结核病的防御机制中，没有一氧化氮(NO)和活性氮中间体(RNI)。我们的研究发现，CTP F基因可能在很大程度上参与了结核杆菌的这种防御机制的逃逸机制。我们这项研究的目的是证实这一结果，并阐明这些基因的具体功能。采用等位基因互换的方法敲除CTP F基因，获得基因缺失菌株。我们在体外检测了NO和RNI揭示的敲除菌株的影响，并将其与野生菌株进行了比较。此外，我们让这些敲除菌株感染人THP-1巨噬细胞，并将敲除菌株与野生菌株进行比较，并检测活细菌的数量、细胞的存活或死亡以及随着时间的推移对细胞凋亡的指示。在未来，我们将用小鼠进行活体实验。

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.