Guerra Susana, López-Fernández Luis A, Conde Raquel, Pascual-Montano Alberto, Harshman Keith, Esteban Mariano
Centro Nacional de Biotecnología, CSIC, Campus Universidad Autónoma, 28049 Madrid, Spain.
J Virol. 2004 Jun;78(11):5820-34. doi: 10.1128/JVI.78.11.5820-5834.2004.
The potential use of the modified vaccinia virus Ankara (MVA) strain as a live recombinant vector to deliver antigens and elicit protective immune responses against infectious diseases demands a comprehensive understanding of the effect of MVA infection on human host gene expression. We used microarrays containing more than 15,000 human cDNAs to identify gene expression changes in human HeLa cell cultures at 2, 6, and 16 h postinfection. Clustering of the 410 differentially regulated genes identified 11 discrete gene clusters with altered expression patterns after MVA infection. Clusters 1 and 2 (accounting for 16.59% [68 of 410] of the genes) contained 68 transcripts showing a robust induction pattern that was maintained during the course of infection. Changes in cellular gene transcription detected by microarrays after MVA infection were confirmed for selected genes by Northern blot analysis and by real-time reverse transcription-PCR. Upregulated transcripts in clusters 1 and 2 included 20 genes implicated in immune responses, including interleukin 1A (IL-1A), IL-6, IL-7, IL-8, and IL-15 genes. MVA infection also stimulated the expression of NF-kappaB and components of the NF-kappaB signal transduction pathway, including p50 and TRAF-interacting protein. A marked increase in the expression of histone family members was also induced during MVA infection. Expression of the Wiskott-Aldrich syndrome family members WAS, WASF1, and the small GTP-binding protein RAC-1, which are involved in actin cytoskeleton reorganization, was enhanced after MVA infection. This study demonstrates that MVA infection triggered the induction of groups of genes, some of which may be involved in host resistance and immune modulation during virus infection.
改良安卡拉痘苗病毒(MVA)株作为一种活重组载体用于递送抗原并引发针对传染病的保护性免疫反应,这需要全面了解MVA感染对人类宿主基因表达的影响。我们使用包含超过15,000个人类cDNA的微阵列来鉴定感染后2小时、6小时和16小时人HeLa细胞培养物中的基因表达变化。对410个差异调节基因进行聚类分析,确定了11个离散的基因簇,其在MVA感染后表达模式发生改变。簇1和簇2(占基因总数的16.59%[410个中的68个])包含68个转录本,显示出在感染过程中持续的强烈诱导模式。通过Northern印迹分析和实时逆转录PCR对选定基因进行验证,证实了MVA感染后通过微阵列检测到的细胞基因转录变化。簇1和簇2中上调的转录本包括20个与免疫反应相关的基因,包括白细胞介素1A(IL-1A)、IL-6、IL-7、IL-8和IL-15基因。MVA感染还刺激了NF-κB及其信号转导途径成分的表达,包括p50和TRAF相互作用蛋白。在MVA感染期间,组蛋白家族成员的表达也显著增加。参与肌动蛋白细胞骨架重组的威斯科特-奥尔德里奇综合征家族成员WAS、WASF1和小GTP结合蛋白RAC-1的表达在MVA感染后增强。本研究表明,MVA感染触发了基因群的诱导,其中一些基因可能参与病毒感染期间的宿主抗性和免疫调节。