Host response to the attenuated poxvirus vector NYVAC:: Upregulation of apoptotic genes and NF-κB-responsive genes in infected HeLa cells
Host response to the attenuated poxvirus vector NYVAC:: Upregulation of apoptotic genes and NF-κB-responsive genes in infected HeLa cells
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DOI：
10.1128/jvi.80.2.985-998.2006
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发表时间：
2006-01-01
影响因子：
5.4
通讯作者：
Esteban, M
中科院分区：
文献类型：
--
作者：
Guerra, S;López-Fernández, LA;Esteban, M
NYVAC已被设计为一种安全、减毒的痘苗病毒(W)载体，用于对广泛范围的病原体和肿瘤进行疫苗接种。由于基于NYVAC的载体作为疫苗的兴趣，以及目前该载体的I/II期临床试验，有必要分析人类对NYVAC感染的宿主反应。利用高密度基因芯片，我们发现了368个NYVAC感染HeLa细胞后的差异表达基因。对受调控基因的聚类确定了六个表达模式发生变化的离散基因簇。I~3类基因占调控基因的47.5%，具有3种类型的基因激活动力学，而4~6类基因则表现出明显的阻遏动力学。对选定基因的实时定量逆转录-聚合酶链式反应分析验证了阵列数据。上调的转录本与免疫反应中涉及的基因相关，包括那些编码白细胞介素1受体2(IL-IR2)、IL-6、ISG-15、CD-80和TNFSF7的基因。NYVAC上调了包括caspase-9在内的几个凋亡级联中间产物的表达，这与其诱导细胞凋亡的能力有关。NYVAC感染也刺激了核因子-kappaB和核因子-kappaB的表达以及核因子-kappaB靶基因的表达。在NYVAC感染过程中，W宿主范围K1L基因的表达阻止了NF-kappa B的激活，但不能诱导细胞凋亡。这项研究是首次全面分析人类细胞对NYVAC感染的转录反应，并为未来评估该载体及其衍生物作为人类疫苗的功能研究提供了一个框架。
NYVAC has been engineered as a safe, attenuated vaccinia virus (W) vector for use in vaccination against a broad spectrum of pathogens and tumors. Due to the interest in NYVAC-based vectors as vaccines and current phase I/II clinical trials with this vector, there is a need to analyze the human host response to NYVAC infection. Using high-density cDNA microarrays, we found 368 differentially regulated genes after NYVAC infection of HeLa cells. Clustering of the regulated genes identified six discrete gene clusters with altered expression patterns. Clusters I to 3 represented 47.5% of the regulated genes, with three patterns of gene activation kinetics, whereas clusters 4 to 6 showed distinct repression kinetics. Quantitative real-time reverse transcription-PCR analysis of selected genes validated the array data. Upregulated transcripts correlated with genes implicated in immune responses, including those encoding interleukin-1 receptor 2 (IL-IR2), IL-6, ISG-15, CD-80, and TNFSF7. NYVAC upregulated several intermediates of apoptotic cascades, including caspase-9, correlating with its ability to induce apoptosis. NYVAC infection also stimulated the expression of NF-kappa B1 and NF-kappa B2 as well as that of NF-kappa B target genes. Expression of the W host range K1L gene during NYVAC infection prevented NF-kappa B activation, but not the induction of apoptosis. This study is the first overall analysis of the transcriptional response of human cells to NYVAC infection and provides a framework for future functional studies to evaluate this vector and its derivatives as human vaccines.