Differentially expressed genes during spontaneous lytic switch of Marek's disease virus in lymphoblastoid cell lines determined by global gene expression profiling.
Differentially expressed genes during spontaneous lytic switch of Marek's disease virus in lymphoblastoid cell lines determined by global gene expression profiling.
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DOI：
10.1099/jgv.0.000744
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发表时间：
2017-04
期刊：
影响因子：
--
通讯作者：
Nair V
中科院分区：
文献类型：
--
作者：
Mwangi WN;Vasoya D;Kgosana LB;Watson M;Nair V
马立克氏病病毒（Marek's disease virus，MDV）是一种感染家禽的α疱疹病毒，可引起马立克氏病（Marek's disease，MD），在易感宿主中以内脏CD 4 +TCRαβ+ T细胞淋巴瘤为特征。已经从MD肿瘤的离体培养物中产生了携带病毒基因组的永生细胞系。MDV转化的类淋巴母细胞系作为一种可获得大量细胞的来源，在研究病毒-宿主相互作用方面具有重要价值。虽然大多数细胞中的病毒基因组处于潜伏状态，但少数细胞群体显示可通过裂解病毒基因的表达识别的自发再活化。这些细胞中的自发再活化提供了研究病毒再活化所涉及的生物学过程的机会。为了详细表征与再活化相关的分子事件，我们使用了两种来源于pRB 1B-UL 47 eGFP诱导的淋巴瘤的淋巴母细胞样细胞系，pRB 1B-UL 47 eGFP是一种重组MDV，经工程改造以表达与UL 47融合的增强型绿色荧光蛋白（EGFP）。我们使用荧光激活细胞分选来从大多数EGFP阴性细胞中纯化具有自发激活病毒基因组的低频EGFP阳性细胞，并使用Illumina HiSeq 2500通过RNA-seq分析其基因表达谱。对裂解感染（EGFP阳性）和潜伏感染（EGFP阴性）细胞群体之间2000多个差异表达基因的免疫途径分析确定了参与再激活的生物学途径。病毒再活化细胞表现出大量病毒基因的差异表达，表达水平存在层次差异。在经历裂解转换的LCL中也注意到许多宿主基因的下调，包括那些直接参与T细胞活化的基因，如CD 3、CD 28、ICOS和磷脂酶C。
Marek’s disease virus (MDV), an alphaherpesvirus of poultry, causes Marek’s disease and is characterized by visceral CD4+TCRαβ+ T-cell lymphomas in susceptible hosts. Immortal cell lines harbouring the viral genome have been generated from ex vivo cultures of MD tumours. As readily available sources of large numbers of cells, MDV-transformed lymphoblastoid cell lines (LCLs) are extremely valuable for studies of virus–host interaction. While the viral genome in most cells is held in a latent state, minor populations of cells display spontaneous reactivation identifiable by the expression of lytic viral genes. Spontaneous reactivation in these cells presents an opportunity to investigate the biological processes involved in the virus reactivation. For detailed characterization of the molecular events associated with reactivation, we used two lymphoblastoid cell lines derived from lymphomas induced by pRB1B-UL47eGFP, a recombinant MDV engineered to express enhanced green fluorescent protein (EGFP) fused with the UL47. We used fluorescence-activated cell sorting to purify the low-frequency EGFP-positive cells with a spontaneously activating viral genome from the majority EGFP-negative cells and analysed their gene expression profiles by RNA-seq using Illumina HiSeq2500. Ingenuity pathway analysis on more than 2000 differentially expressed genes between the lytically infected (EGFP-positive) and latently infected (EGFP-negative) cell populations identified the biological pathways involved in the reactivation. Virus-reactivating cells exhibited differential expression of a significant number of viral genes, with hierarchical differences in expression levels. Downregulation of a number of host genes including those directly involved in T-cell activation, such as CD3, CD28, ICOS and phospholipase C, was also noticed in the LCL undergoing lytic switch.
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DOI：
10.1093/bioinformatics/btu638
发表时间：
2015-01-15
期刊：
Bioinformatics (Oxford, England)
影响因子：
--
作者：
Anders S;Pyl PT;Huber W
通讯作者：
Huber W
影响因子：
3.7
作者：
Hunt, HD;Lupiani, B;Parcells, MS
通讯作者：
Parcells, MS
影响因子：
5.4
作者：
DELECLUSE, HJ;HAMMERSCHMIDT, W
通讯作者：
HAMMERSCHMIDT, W
DOI：
10.1073/pnas.0404508101
发表时间：
2004-08-10
影响因子：
11.1
作者：
Lupiani, B;Lee, LF;Reddy, SM
通讯作者：
Reddy, SM
影响因子：
15.3
作者：
Kaufer, Benedikt B.;Jarosinski, Keith W.;Osterrieder, Nikolaus
通讯作者：
Osterrieder, Nikolaus