Chlamydia pneumoniae persistent infection and its mechanism

目前的研究表明，专性细胞内细菌衣原体肺炎不仅与呼吸系统疾病有关，而且与动脉粥样硬化等慢性疾病有关。在心血管疾病患者的冠状动脉粥样硬化斑块和外周血中检测到肺炎支原体DNA或抗原，增强了其参与动脉粥样硬化发病的可能性。此外，一些研究表明，通过培养和RT-PCR方法，可以在老年冠心病患者的动脉粥样硬化斑块中检测到活的肺炎衣原体。我们最近的研究还表明，从健康供者获得的外周血单个核细胞（PBMCs）中很容易检测到活菌肺炎球菌，这表明血液中活菌的存在可能是动脉粥样硬化发展的一个危险因素。肺炎梭菌优先感染呼吸道上皮细胞和巨噬细胞。另一方面，我们之前的研究表明，淋巴细胞是另一种宿主细胞，允许肺炎梭菌感染具有持续性。由于淋巴细胞是动脉粥样硬化发生过程中除巨噬细胞外的主要免疫细胞类型，因此淋巴细胞与该病原体的相互作用可能参与了肺炎原体相关慢性炎症性疾病的发病机制。我们首先试图评估非肥胖糖尿病（NOD）小鼠的糖尿病发展与肺炎原体从肺部传播到外周血之间的可能关联。采用实时逆转录聚合酶链反应（RT-PCR），在多次鼻内接种肺炎C. 16S rRNA引物后，在患有糖尿病的NOD小鼠（38.5%）和ICR小鼠（40%）中检测到肺部细菌，而在没有糖尿病的NOD小鼠（糖尿病前期NOD小鼠和非糖尿病退休NOD小鼠）中检测到细菌的比例很低（4.8%）。该细菌仅在糖尿病NOD小鼠的外周血单个核细胞（PBMCs）中检测到（53.8%）。异硫氰酸荧光素偶联抗衣原体单克隆抗体免疫染色结果也显示肺中存在细菌抗原，RT-PCR判断PBMCs阳性。结果清楚地表明，NOD小鼠糖尿病的发生可能是促进肺炎原体从肺向外周血传播的关键因素之一。其次，我们检测了肺炎球菌感染对人淋巴细胞（Molt-4细胞和外周血单核细胞（PBMCs）富集淋巴细胞）CD3和CD25表达的可能改变。肺炎衣原体感染后，Molt-4细胞中这两个分子的表达水平均显著降低。临床分离的肺炎球菌也引起了感染细胞的两种表达的改变。相比之下，沙眼衣原体未引起Molt-4细胞两种表达的改变。热灭菌和HEp-2裂解物均未引起淋巴细胞CD3表达的改变。在培养液中加入NS-398 （Cox-2抑制剂）或AH-23848 （EP4前列腺素受体拮抗剂）可消除感染细胞CD3表达的改变。竞争酶联免疫吸附试验也证实了感染细胞培养上清中前列腺素E_2 （PGE_2）的增加。从mpbmcs富集的淋巴细胞感染肺炎c也引起CD3表达的改变。因此，肺炎衣原体感染人淋巴细胞可诱导PGE_2产生介导的CD3表达改变。由于CD3分子在抗原识别后的信号转导中起主要作用，因此这种改变可能与肺炎梭菌持续感染淋巴细胞有关。此外，我们发现2-氨基-吩恶嗪3- 1（吩恶嗪衍生物：Phx-3）抑制肺炎球菌在人类单核细胞和上皮细胞中的复制，部分依赖于宿主细胞的色氨酸代谢途径。少

Current studies have revealed that the obligate intracellular bacterium Chlamydia (Chlamydophila) pneumoniae is associated not only with respiratory diseases, but also with chronic diseases such as atherosclerosis. The detection of the C. pneumoniae DNA or antigen in coronary atherosclerotic plaque and peripheral blood of the patients with cardiovascular diseases has strengthened the likelihood of its possible involvement in the pathogenesis of atherosclerosis. Moreover, several studies indicate that viable C. pneumoniae can be detected in atherosclerotic plaques from patients of advanced age with coronary heart disease by both culture and RT-PCR methods. Our recent studies also showed that viable C. pneumoniae are readily detectable in peripheral blood mononuclear cells (PBMCs) obtained from healthy donors, indicating that the presence of viable bacteria in the blood stream might be a risk factor for the development of atherosclerosis. C. pneumoniae preferentially infects respiratory … More tract epithelial cells as well as macrophages. On the other hand, our previous studies revealed that lymphocyte is another host cell permitting C. pneumoniae infection with a persistent nature. Since lymphocytes are a major immune cell type besides macrophages in the development of atherosclerosis, interaction between lymphocytes and this pathogen may contribute to the pathogenesis of chronic inflammatory diseases associated with C. pneumoniae. We first attempted to assess a possible association between development of diabetes in non-obese diabetic (NOD) mice and dissemination of C. pneumoniae from lung to peripheral blood. By real-time reverse transcription-polymerase chain reaction (RT-PCR) with primers for C. pneumoniae 16S rRNA, following multiple intranasal inoculations,. bacteria in lung were detected in NOD mice with diabetes (38.5 %) as well as ICR mice (40 %), but prevalence of bacteria in NOD mice without diabetes (Pre-diabetic NOD mice and non-diabetic retired NOD mice) was very low (4.8 %).The bacteria were only detected in peripheral blood mononuclear cells (PBMCs) cultured withhydrocortisone of the NOD mice with diabetes (53.8 %). Results of immunostaining with fluorescein isothiocyanate-conjugated anti-chlamydia monoclonal antibody also showed the presence of bacterial antigens m the lungs and the PBMCs judged as positive by the RT-PCR. The results clearly showed that the development of diabetes in NOD mouse appears to be one of critical factors for promoting the dissemination of C. pneumoniae from lung to peripheral blood. Second, we examined a possible alteration of CD3 and CD25 expressions of human lymphocytes [Molt-4 cells and enriched lymphocytes from peripheral blood mononuclear cells (PBMCs)] by C. pneumoniae infection. The expression levels of both molecules of Molt-4 cells were significantly decreased by C. pneumoniae infection. C. pneumoniae clinical isolates also caused the alteration of both expressions of the infected cells. In contrast, C. trachomatis did not cause any alteration of both expressions of Molt-4 cells. Heat-killed bacteria and HEp-2 lysate as mock did not cause any alteration of CD3 expression of lymphocytes. Addition of either NS-398 (Cox-2 inhibitor) or AH-23848 (EP4 prostanoid receptorantagonist) tothe culture abolishedthe alteration of CD3 expression ofthe infected cells. The enhanced prostaglandin E_2 (PGE_2) productions in the culture supernatant of infected cells were also confirmed by competitiveELISA. C. pneumoniae infection of enriched lymphocytesfromPBMCs also causedanalteration of CD3 expression.Thus, C. pneumoniae infection of human lymphocytes induces an alteration of CD3 expression mediated by PGE_2 production. Since CD3 molecule has a major role in the signal transduction following antigen recognitions, such alteration may be involved in C.pneumoniae persistent infection of lymphocytes. Also, we foundthat 2-amino-phenoxazine 3-one (phenoxazine derivate: Phx-3)inhibits C. pneumoniae replication in human monocytic cells as well as epithelial cells, partially depending on the tryptophan-metabolic pathway of host cells. Less