Ren Qun, Robertson Shelly J, Howe Dale, Barrows Lorraine F, Heinzen Robert A
Department of Molecular Biology, University of Wyoming, Laramie, Wyoming 82071-3944, USA.
Ann N Y Acad Sci. 2003 Jun;990:701-13. doi: 10.1111/j.1749-6632.2003.tb07447.x.
DNA microarray analysis was conducted to investigate the transcriptional responses of the human monocytic leukemia cell line THP-1 to infection by Chlamydia trachomatis or Coxiella burnetii. RNA was isolated from mock infected cells and cells infected for 36 hours using TRIzol reagent. Biotinylated probes synthesized from RNA samples were hybridized to an Affymetrix U133A human genome chip consisting of 18,462 human gene probe sets. A total of 335 and 548 THP-1 genes were up- or downregulated at least twofold in cells infected with C. burnetii or C. trachomatis, respectively, when compared to uninfected cells. There was a high degree of overlap in transcriptional responses to infection with shared responses observed for 39 downregulated and 189 upregulated genes. Numerous pathogen-specific transcriptional responses were also observed. Quantitative RT-PCR and immunoblotting confirmed up- or down-regulation of a subset of THP-1 genes in response to infection by C. burnetii. This study provides insight into the host transcriptional responses to infection by Chlamydia and Coxiella that may affect the infectious cycle of each organism.
进行DNA微阵列分析以研究人单核细胞白血病细胞系THP-1对沙眼衣原体或贝氏柯克斯体感染的转录反应。使用TRIzol试剂从模拟感染细胞和感染36小时的细胞中分离RNA。从RNA样本合成的生物素化探针与由18,462个人类基因探针集组成的Affymetrix U133A人类基因组芯片杂交。与未感染细胞相比，在感染贝氏柯克斯体或沙眼衣原体的细胞中，分别有335个和548个THP-1基因上调或下调至少两倍。在对感染的转录反应中存在高度重叠，观察到39个下调基因和189个上调基因有共同反应。还观察到许多病原体特异性转录反应。定量RT-PCR和免疫印迹证实了THP-1基因的一个子集在贝氏柯克斯体感染后上调或下调。本研究深入了解了宿主对衣原体和柯克斯体感染的转录反应，这些反应可能会影响每种生物体的感染周期。
