Lewis Maria E, Belland Robert J, AbdelRahman Yasser M, Beatty Wandy L, Aiyar Ashok A, Zea Arnold H, Greene Sheila J, Marrero Luis, Buckner Lyndsey R, Tate David J, McGowin Chris L, Kozlowski Pamela A, O'Brien Michelle, Lillis Rebecca A, Martin David H, Quayle Alison J
Department of Microbiology, Immunology and Parasitology, Louisiana State University Health Sciences Center New Orleans, LA, USA.
Department of Microbiology, Immunology and Biochemistry, University of Tennessee Health Sciences Center Memphis, TN, USA.
Front Cell Infect Microbiol. 2014 Jun 10;4:71. doi: 10.3389/fcimb.2014.00071. eCollection 2014.
In vitro models of Chlamydia trachomatis growth have long been studied to predict growth in vivo. Alternative or persistent growth modes in vitro have been shown to occur under the influence of numerous stressors but have not been studied in vivo. Here, we report the development of methods for sampling human infections from the endocervix in a manner that permits a multifaceted analysis of the bacteria, host and the endocervical environment. Our approach permits evaluating total bacterial load, transcriptional patterns, morphology by immunofluorescence and electron microscopy, and levels of cytokines and nutrients in the infection microenvironment. By applying this approach to two pilot patients with disparate infections, we have determined that their contrasting growth patterns correlate with strikingly distinct transcriptional biomarkers, and are associated with differences in local levels of IFNγ. Our multifaceted approach will be useful to dissect infections in the human host and be useful in identifying patients at risk for chronic disease. Importantly, the molecular and morphological analyses described here indicate that persistent growth forms can be isolated from the human endocervix when the infection microenvironment resembles the in vitro model of IFNγ-induced persistence.
长期以来，沙眼衣原体生长的体外模型一直被用于预测其在体内的生长情况。体外的替代或持续生长模式已被证明在众多应激源的影响下会出现，但尚未在体内进行研究。在此，我们报告了从子宫颈采集人类感染样本的方法的开发，该方法允许对细菌、宿主和子宫颈环境进行多方面分析。我们的方法能够评估细菌总载量、转录模式、通过免疫荧光和电子显微镜观察的形态，以及感染微环境中细胞因子和营养物质的水平。通过将这种方法应用于两名感染情况不同的试点患者，我们确定他们截然不同的生长模式与截然不同的转录生物标志物相关，并与局部干扰素γ水平的差异有关。我们的多方面方法将有助于剖析人类宿主中的感染情况，并有助于识别有患慢性病风险的患者。重要的是，此处描述的分子和形态学分析表明，当感染微环境类似于干扰素γ诱导的持续性体外模型时，可以从人类子宫颈中分离出持续生长形式。
