Dallo S F, Baseman J B
Department of Microbiology, University of Texas Health Science Center at San Antonio, 7703 Floyd Curl Drive, San Antonio, TX 78229-3900, USA.
Microb Pathog. 2000 Nov;29(5):301-9. doi: 10.1006/mpat.2000.0395.
We examined intracellular survival and growth of pathogenic mycoplasmas (Mycoplasma penetrans, Mycoplasma pneumoniae and Mycoplasma genitalium) in cultured human cells. By using the eukaryotic nuclear DNA synthesis inhibitor, aphidicolin, we detected the selective synthesis of mycoplasma (My) and mitochondria (Mt) DNA, which could be further differentiated by restriction enzyme analyses. Also, intracellular M. pneumoniae and M. penetrans infectivity of human cells was detected over 6 months using subfractionation of infected cells and determination of mycoIplasma colony forming units (cfu). For M. genitalium, which we failed to re-grow from infected cells, species-specific PCR primers were used to implicate long-term mycoplasma survivability. Data indicated that pathogenic mycoplasmas reside and replicate intracellularly over extended periods in human cells, consistent with the ability of mycoplasmas to circumvent antibiotic therapy and immune surveillance and establish chronic infections.
我们检测了致病性支原体（穿透支原体、肺炎支原体和生殖支原体）在培养的人细胞中的细胞内存活和生长情况。通过使用真核细胞核DNA合成抑制剂阿非迪霉素，我们检测到了支原体（My）和线粒体（Mt）DNA的选择性合成，这可以通过限制性内切酶分析进一步区分。此外，通过对感染细胞进行亚分级分离并测定支原体集落形成单位（cfu），在6个月以上的时间里检测了人细胞内肺炎支原体和穿透支原体的感染性。对于我们未能从感染细胞中重新培养出来的生殖支原体，使用物种特异性PCR引物来证明支原体的长期生存能力。数据表明，致病性支原体在人细胞内长期存在并复制，这与支原体规避抗生素治疗和免疫监视并建立慢性感染的能力相一致。
