Chlamydia pneumoniae dissemination and atherosclerosis

肺炎衣原体传播和动脉粥样硬化

• 批准号: 15590399
• 负责人: YAMAGUCHI Hiroyuki
• 金额: $ 2.3万
• 依托单位: Osaka University (2004)Kyorin University (2003)
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15590399/
• 关键词: Chlamydia pneumoniae; PBMC; dissemination; Real-time RT-PCR; Atherosclerosis; NOD mice; Animal model; real-time RT-PCR法; ルイスラット; 末梢血液; 生体内伝播; クラミジア・ニューモニエ; 未梢血液PBMC; real-time PCR; Syber green; 動物モデル; 生菌; RNA

Current studies have revealed that the obligate intracellular bacterium Chlamydia (Chlamydophila) pneumoniae is associated not only with respiratory diseases but also with atherosclerosis. In the present study, to understand the viability and growth potential of C.pneumoniae detected in blood of healthy persons, we first attempted to detect C.pneumoniae transcript in peripheral blood mononuclear cells (PBMCs) obtained from 70 healthy donors by real-time RT-PCR method. The presence of C.pneumoniae transcript in PBMCs from blood of healthy human donors was assessed by real-time RT-PCR-using primers for C.pneumoniae 16S rRNA. Out of 70 donors (18.5 %) showed the presence of bacterial transcript in cultured PBMCs. Prevalence of bacterial detection and bacterial numbers were significantly increased in PBMNC cultures incubated with cycloheximide. These results showed that viable C.pneumoniae might be present in healthy human PBMCs. Although several studies reported C.pneumoniae systemic diss … More emination from lung in animal model by DNA-based qualitative methods, the lack of a suitable animal model for permitting viable C.pneumonia dissemination from lung to blood has impeded the understanding of how viable C.pneumoniae can reach blood vessels and trigger the development of atherosclerosis. Therefore, we next studied the susceptibility of non-obese diabetic (NOD) mice to dissemination of viable C.pneumoniae from lung to blood, as detected by real-time RT-PCR. Following multiple intranasal inoculations, bacteria in lung were detected in NOD mice with diabetes (38.5%, n=13) as well as ICR mice (40%, n=10), but prevalence of bacteria in NOD mice without diabetes [Pre-diabetic NOD mice (n=8) and non-diabetic retired NOD mice (n=13)] was very low (5.2%, n=21). The bacteria were only detected in cultured PBMCs (53.8 %) of the NOD mice with diabetes. The prevalence of bacterial detection appeared to be increased in PBMC cultures incubated with hydrocortisone. Cultivation of C.pneumoniae from cultured PBMCs from all NOD mice was unsuccessful. Results of immunostaining with fluorescein isothiocyanate-conjugated anti-Chlamydia monoclonal antibody also showed the presence of bacterial antigens in the lungs and the PBMCs judged as positive by the RT-PCR. Thus, diabetic NOD mouse is sensitive to C.pneumoniae infection in which a possible dissemination of viable bacteria from lung to blood, and the animal model established in this study may be useful for understanding dissemination of C.pneumoniae infection. Less

目前的研究表明，专性细胞内细菌肺炎衣原体不仅与呼吸道疾病有关，而且与动脉粥样硬化有关。在本研究中，为了了解健康人血液中检测到的肺炎衣原体的活力和生长潜力，我们进行了研究。首先尝试通过实时RT-PCR方法检测从70名健康捐献者获得的外周血单核细胞（PBMC）中肺炎衣原体转录物的存在。使用肺炎衣原体 16S rRNA 引物，通过实时 RT-PCR 评估健康人类供体血液中的肺炎衣原体转录物，70 名供体 (18.5%) 中显示培养的 PBMC 中存在细菌转录物。在用放线菌酮培养的 PBMNC 培养物中，细菌检测和细菌数量显着增加。这些结果表明，健康人体内可能存在活的肺炎衣原体。尽管一些研究报告了肺炎衣原体通过基于 DNA 的定性方法从动物模型中的肺部排出，但缺乏合适的动物模型来允许肺炎衣原体从肺部传播到血液中，这阻碍了对肺炎衣原体如何传播的理解。活的肺炎衣原体可以到达血管并引发动脉粥样硬化的发展，因此，我们接下来研究非肥胖糖尿病（NOD）小鼠对肺炎衣原体传播的敏感性。通过实时 RT-PCR 检测，在多次鼻内接种后，在患有糖尿病的 NOD 小鼠（38.5%，n=13）和 ICR 小鼠（40%）中检测到肺部存在活肺炎衣原体。 ，n = 10），但没有糖尿病的 NOD 小鼠[糖尿病前期 NOD 小鼠（n = 8）和非糖尿病退役 NOD 小鼠（n = 13）]中的细菌流行率非常低(5.2%，n=21)。仅在患有糖尿病的 NOD 小鼠的培养 PBMC 中检测到细菌。在用肺炎衣原体培养的 PBMC 培养物中，细菌检测的发生率似乎有所增加。所有 NOD 小鼠培养的 PBMC 均未成功使用异硫氰酸荧光素偶联的抗衣原体单克隆抗体进行免疫染色。抗体还显示肺部和 PBMC 中存在细菌抗原，经 RT-PCR 判断为阳性。因此，糖尿病 NOD 小鼠对肺炎衣原体感染敏感，其中活细菌可能从肺部传播到血液，并且糖尿病 NOD 小鼠对肺炎衣原体感染敏感。本研究建立的动物模型可能有助于了解肺炎衣原体感染的传播。

项目成果

Yamaguchi H. et al.: "Chlamydia pneumoniae resists antibiotics in lymphocytes."Antimicrobial Agents and Chemotherapy. 47(6). 1972-1975 (2003)

Yamaguchi H.等人：“肺炎衣原体对淋巴细胞中的抗生素具有抵抗力。”抗菌药物和化疗。

Prevalence of viable Chlamydia pneumoniae in peripheral blood mononuclear cells of healthy blood donors.

健康献血者外周血单核细胞中活肺炎衣原体的患病率。

• 发表时间: 2004
• 期刊: Transfusion 44
• 作者: Takahashi M;Taguchi H;Yamaguchi H;Osaki T;Komatsu A;Kamiya S.;Hiroyuki Yamaguchi et al.
• 通讯作者: Hiroyuki Yamaguchi et al.

Matsnaga K et al.: "Legionella pneumophila suppresses macrophage interleukin-12 production by activating the p42/44 mitogen-activated preotein kinase cascade."Infection and Immunity. 71(11). 6672-6675 (2003)

Matsnaga K 等人：“嗜肺军团菌通过激活 p42/44 丝裂原激活的蛋白激酶级联来抑制巨噬细胞白细胞介素 12 的产生。”感染和免疫。

Yamaguchi H. et al.: "Prevalence of viable Chlamydia pneumoniae in PBMNCs of Healthy donors."Transfusion. in press. (2004)

Yamaguchi H. 等人：“健康捐献者的 PBMNC 中存活的肺炎衣原体的患病率。”输血。

Chlamydia pneumoniae resists antibiotics in lymphocytes.

肺炎衣原体对淋巴细胞中的抗生素具有抵抗力。

• 发表时间: 2003
• 期刊: Antimicrobial Agents and Chemotherapy 47
• 作者: Takahashi M;Taguchi H;Yamaguchi H;Osaki T;Komatsu A;Kamiya S.;Hiroyuki Yamaguchi et al.;Motomichi Takahashi et al.;Yamaguchi H. et al.;Yamaguchi H. et al.
• 通讯作者: Yamaguchi H. et al.