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猪弓形虫感染体外模型中宿主-病原体相互作用的cDNA微阵列分析

cDNA microarray analysis of host-pathogen interactions in a porcine in vitro model for Toxoplasma gondii infection.

作者信息

Okomo-Adhiambo Margaret, Beattie Craig, Rink Anette

机构信息

Department of Animal Biotechnology, University of Nevada, 1664 North Virginia Street, Reno, NV 89557, USA.

出版信息

Infect Immun. 2006 Jul;74(7):4254-65. doi: 10.1128/IAI.00386-05.

Abstract

Toxoplasma gondii induces the expression of proinflammatory cytokines, reorganizes organelles, scavenges nutrients, and inhibits apoptosis in infected host cells. We used a cDNA microarray of 420 annotated porcine expressed sequence tags to analyze the molecular basis of these changes at eight time points over a 72-hour period in porcine kidney epithelial (PK13) cells infected with T. gondii. A total of 401 genes with Cy3 and Cy5 spot intensities of >/=500 were selected for analysis, of which 263 (65.6%) were induced >/=2-fold (expression ratio, >/=2.0; P </= 0.05 [t test]) over at least one time point and 48 (12%) were significantly down-regulated. At least 12 functional categories of genes were modulated (up- or down-regulated) by T. gondii. The majority of induced genes were clustered as transcription, signal transduction, host immune response, nutrient metabolism, and apoptosis related. The expression of selected genes altered by T. gondii was validated by quantitative real-time reverse transcription-PCR. These results suggest that significant changes in gene expression occur in response to T. gondii infection in PK13 cells, facilitating further analysis of host-pathogen interactions in toxoplasmosis in a secondary host.

摘要

刚地弓形虫可诱导促炎细胞因子的表达,使细胞器重新排列,清除营养物质,并抑制受感染宿主细胞的凋亡。我们使用了一个包含420个注释猪表达序列标签的cDNA微阵列,来分析猪肾上皮(PK13)细胞在感染刚地弓形虫后72小时内8个时间点这些变化的分子基础。总共选择了401个Cy3和Cy5斑点强度≥500的基因进行分析,其中263个(65.6%)在至少一个时间点上被诱导上调≥2倍(表达比≥2.0;P≤0.05 [t检验]),48个(12%)被显著下调。刚地弓形虫调节(上调或下调)了至少12个功能类别的基因。大多数诱导基因聚类为与转录、信号转导、宿主免疫反应、营养代谢和凋亡相关的基因。通过定量实时逆转录PCR验证了刚地弓形虫改变的选定基因的表达。这些结果表明,PK13细胞在感染刚地弓形虫后基因表达发生了显著变化,有助于进一步分析中间宿主弓形虫病中宿主-病原体的相互作用。

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