Gérard H C, Köhler L, Branigan P J, Zeidler H, Schumacher H R, Hudson A P
Department of Immunology and Microbiology, Wayne State University School of Medicine, Detroit, Michigan 48201, USA.
Med Microbiol Immunol. 1998 Oct;187(2):115-20. doi: 10.1007/s004300050082.
The principal host cell for persistently infecting synovial Chlamydia trachomatis is the macrophage. During infection of human monocytes/macrophages in culture this bacterium displays aberrant morphology and produces no new elementary bodies, reflecting the situation in synovium. Here we investigate the metabolic status of C. trachomatis (serovar K) during an extended infection of human peripheral monocytes in vitro. Using reverse transcription-polymerase chain reaction assays, we have shown that primary transcripts from the chlamydial rRNA operons are present throughout a 10-day course of infection. Other assays targeting mRNAs from chlamydial genes encoding r-proteins S5 and L5, the glycyl-tRNA synthetase, the 60-kDa cysteine-rich outer membrane protein, and the KDO transferase indicate that these messengers are also present throughout the entire 10-day period. The gene encoding the 57-kDa heat-shock protein (hsp60) is expressed by the bacterium throughout the 10-day infection of cultured monocytes, but transcript levels from the gene encoding the major outer membrane protein (omp1) appear to be attenuated. Western analyses targeting these latter proteins confirm the presence of the hsp60 gene product, and the virtual absence of major outer membrane protein, in chlamydia-infected cultured human monocytes. Thus, during extended infection of human monocytes in vitro, chlamydia are non-productive but transcriptionally active; the pattern of transcriptional activity reflects that known for persistent C. trachomatis infection in vivo in synovial tissue.
持续感染滑膜沙眼衣原体的主要宿主细胞是巨噬细胞。在体外培养的人单核细胞/巨噬细胞感染过程中,这种细菌呈现异常形态,且不产生新的原体,这反映了滑膜中的情况。在此,我们研究了沙眼衣原体(血清型K)在体外长时间感染人外周单核细胞期间的代谢状态。通过逆转录-聚合酶链反应分析,我们发现衣原体rRNA操纵子的初级转录本在整个10天的感染过程中都存在。针对衣原体基因编码r蛋白S5和L5、甘氨酰-tRNA合成酶、60 kDa富含半胱氨酸的外膜蛋白以及KDO转移酶的mRNA的其他分析表明,这些信使RNA在整个10天期间也都存在。编码57 kDa热休克蛋白(hsp60)的基因在培养的单核细胞10天的感染过程中均由细菌表达,但编码主要外膜蛋白(omp1)的基因的转录水平似乎有所减弱。针对这些后一种蛋白的蛋白质印迹分析证实,在衣原体感染的培养人单核细胞中存在hsp60基因产物,而几乎不存在主要外膜蛋白。因此,在体外人单核细胞的长时间感染过程中,衣原体不具有生产性但转录活跃;转录活性模式反映了滑膜组织中沙眼衣原体体内持续感染的已知情况。