Analysis of the arteriosclerotic change caused by Chlamydia pneumoniae infection in a experimental mouse model (W/BF1)

实验小鼠模型(W/BF1)肺炎衣原体感染引起的动脉硬化变化分析

• 批准号: 09670635
• 负责人: SOEJIMA Rinzo
• 金额: $ 1.86万
• 依托单位: KAWASAKI UNIVERSITY OF MEDICAL WELFARE
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09670635/
• 关键词: Chlamydia.Pneumoniae; (NZWxBXSB)F1 mice; multiple infection model; Direct immunofluorescent antibody test; Immunohistochemical staining; Coronary artery area of section; girth; (NZW×BXSB)F_1マウス; 酵素抗体法; chlamydia pneumoniae; (NZWXBXSB)F_1マウス; 動脈

1.Experiment of single infection : We used (NZWxBXSB)F1. female mice fifty percent of which develop spontaneous myocardial infarction at the age of 24 weeks. Mice were inoculated with C.pneumoniae KKpn-15 strain (1O^5FU/mouse) intranasally at the age of 16 weeks. Mortality rate was 56% at 3days, 100% at 7 days after inoculation. C.pneumoniae was isolated from all the lung tested and histological findings of the lung specimens were compatible with pneumonia. Therefore, it is suggested that C.pneumoniae has a potential to induce lethal pneumonia depending of the host condition.2.Experiment of multiple infections After confirming that lethal pneumonia does not occur at the dose less than 10^4IFU/mouse, mice were inoculated with C.pneumoniae KKpn-15 strain (10^4LFU/mouse) intranasally at the age of 16 weeks. Subsequently, inoculation was repeated 3 times at 35, 42, 49 days after initial inoculation. Three mice were sacrificed at each designated time point till 63 days after initiation and … More we observed chronological change of C.pneumoniae isolation rate and histological change of lung and heart.a)Detection of C.pneumoniae C.pneumoniae was isolated and PCR for C.pneumoniae was positive in lung at 3, 7, 14 post inoculation day respectively. However, it was not detected both in lung and heart thereafter even with multiple inoculation.b)Pathological findings : Transition from acute pulmonary infection to chronic infection after initial inoculation was observed. In heart, no myocardial infarction and arterioscrelotic change was observed.c)Detection by direct immunofluoresent antibody (DFA) test : C.pneumoniae was positively detected in lung at the early phase of infection. But in myocardium and coronary artery, no positive signal was detected at any time point.d)Detection by immunohistochemical staining(ABC method) : No positive staining was observed in myocadrium obtained after multiple inoculation.e)Morphometrical analysis of coronary artery : For the quantitative analysis of intimal thickness of coronary artery, we applied computer-assisted image analysis using an IBAS system. The girth, area of section and ratio of them ( area of section/girth ; S : G ratio) in coronary artery with the inner girth more than 200 mu m were measured. S : G ratio was 16.09 in inoculation group and 16.52 in control group showing no significant difference. Less

1.单次感染实验：我们使用(NZWxBXSB)F1。百分之五十的雌性小鼠在 24 周龄时出现自发性心肌梗塞。小鼠在16周龄时鼻内接种肺炎衣原体KKpn-15株(1O^5FU/小鼠)。接种后3天死亡率为56%，7天死亡率为100%。从所有测试的肺部中分离出肺炎衣原体，并且肺部标本的组织学结果与肺炎相符。因此，提示肺炎衣原体具有诱发致死性肺炎的潜力，具体取决于宿主情况。2.多重感染实验在确认低于10^4IFU/小鼠的剂量下不会发生致死性肺炎后，在16周龄时对小鼠鼻内接种肺炎衣原体KKpn-15株（10^4LFU/小鼠）。随后，在初次接种后35、42、49天重复接种3次。在每个指定时间点处死3只小鼠，直至开始后63天，观察肺炎衣原体分离率的时间变化以及肺和心脏的组织学变化。a)肺炎衣原体的检测，分别在接种后3、7、14天在肺中分离出肺炎衣原体，并且肺炎衣原体PCR呈阳性。然而，即使多次接种，此后在肺和心脏中也未检测到。b)病理结果：观察到初次接种后从急性肺部感染转变为慢性感染。心脏未见心肌梗塞及动脉粥样硬化改变。c)直接免疫荧光抗体(DFA)检测：感染早期肺部检出肺炎衣原体阳性。但在心肌和冠状动脉中，在任何时间点均未检测到阳性信号。 d)免疫组织化学染色检测（ABC法）：多次接种后获得的心肌中未观察到阳性染色。 e)冠状动脉形态分析：对于冠状动脉内膜厚度的定量分析，我们应用IBAS系统进行计算机辅助图像分析。测量内周大于200μm的冠状动脉的周长、截面面积及其比（截面面积/周长；S：G比）。接种组S:G比为16.09，对照组为16.52，无显着性差异。较少的

项目成果

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Takayuki Kano 等人：“感染肺炎衣原体的小鼠的肺和心脏病变”，日本呼吸学会杂志 36（特刊）402（1999 年）。

岸本 寿男 他: "ELISA法による抗Chlamydia pneumoniae特異抗体の測定 1.外膜複合体を用いたELISA法キットの評価" 感染症学雑誌. 70. 821-829 (1996)

Toshio Kishimoto 等人：“通过 ELISA 测量抗肺炎衣原体特异性抗体 1。使用外膜复合物评估 ELISA 试剂盒”《传染病杂志》70. 821-829 (1996)。

Karino T., et al.: "A study of serological prevalence of anti-Chlamydia pneumoniae antibody in hospitalized patients" Journal of the Japanese Respiratory Society.37(suppl). 293 (1999)

Karino T.等人：“住院患者抗肺炎衣原体抗体血清学流行率的研究”日本呼吸学会杂志37（suppl）。

狩野 孝之 他: "当科入院患者におけるChlamydia pneumoniae抗体保有率についての検討" 日本呼吸器学会雑誌. 37(増刊号). 293 (1999)

Takayuki Kano 等：“我科住院患者肺炎衣原体抗体流行情况的研究”，日本呼吸学会杂志 37（特刊）。

Kishimoto T., et al.: "Assay of specific anti-Chlamydia pneumoniae antibodies by ELISA method. 1.Evaluation of ELISA kit using outer membrane complex." Journal of the Japanese Association for Infectious Diseases. 70(8). 821-829 (1996)

Kishimoto T.等人：“通过ELISA方法测定特异性抗肺炎衣原体抗体。1.使用外膜复合物评估ELISA试剂盒。”