Differential gene expression patterns of EBV infected EBNA-3A positive and negative human B lymphocytes.
Differential gene expression patterns of EBV infected EBNA-3A positive and negative human B lymphocytes.
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DOI：
10.1371/journal.ppat.1000506
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发表时间：
2009-07
期刊：
影响因子：
6.7
通讯作者：
Kempkes B
中科院分区：
文献类型：
--
作者：
Hertle ML;Popp C;Petermann S;Maier S;Kremmer E;Lang R;Mages J;Kempkes B
EB病毒(EBV)基因组编码86种蛋白质，但在EB病毒转化的B细胞(LCLS)中只表达一组有限的蛋白质。这些细胞通过EBV核抗原(EBNAs)和潜伏膜蛋白(LMPs)的协同作用进行增殖，其中一些是限速的，以建立稳定的促进生长和抗凋亡活性的动态平衡。我们在这里表明，与EBNA-2缺失的突变病毒相比，缺乏EBNA-3A基因的EBV突变株虽然受损，但仍能启动原代人类B细胞的细胞周期进入和增殖。令人惊讶的是，与以前的报告相反，这些病毒突变在生长转化试验中会减弱，但会导致永久性生长的EBNA-3A阴性B细胞株，表现出增殖率降低和细胞凋亡水平上升。EBNA-3A缺陷LCLS的表达谱由129个下调基因和167个上调基因组成，这些基因显著丰富了参与细胞凋亡过程或细胞周期进展的基因，如肿瘤抑制基因p16/Ink4a，或可能参与感染宿主病毒生命周期的必要步骤。此外，EBNA-3A细胞靶基因与先前确定的EBNA-2靶基因显著重叠。这项研究包括在生长转化的B细胞的病毒蛋白复杂网络中产生的EBNA-3A靶基因的第一个全基因组表达谱，并允许更详细地了解EBNA-3A在病毒致病机制中的功能和贡献。爱泼斯坦-巴尔病毒(EBV)感染原代人类B细胞，并建立潜伏感染，导致B细胞培养永久生长。这些生长转化的B细胞表达一组明确的潜伏病毒基因，这些基因也在免疫抑制患者的移植后淋巴瘤中表达。在协同行动中，这些潜伏的病毒蛋白驱动细胞增殖并防止细胞凋亡。在本研究中，产生了缺乏EB病毒核抗原-3A(EBNA-3A)基因的重组EB病毒突变体。EBNA-3A是基因表达的转录调控因子。在此，我们证明了EBNA-3A缺陷生长转化B细胞可以在体外建立。我们的结果表明，EBNA-3A支持活性，但对感染的B细胞的增殖并不是绝对必要的。借助于已建立的EBNA-3A缺陷细胞系，我们首次在EBV感染的B细胞中鉴定了由EBNA-3A控制的广泛的细胞靶基因。这些EBNA-3A靶基因将使人们能够更详细地了解EBNA-3A在病毒发病机制中的功能和贡献。
The genome of Epstein-Barr virus (EBV) encodes 86 proteins, but only a limited set is expressed in EBV–growth transformed B cells, termed lymphoblastoid cell lines (LCLs). These cells proliferate via the concerted action of EBV nuclear antigens (EBNAs) and latent membrane proteins (LMPs), some of which are rate limiting to establish a stable homeostasis of growth promoting and anti-apoptotic activities. We show here that EBV mutants, which lack the EBNA-3A gene, are impaired but can still initiate cell cycle entry and proliferation of primary human B cells in contrast to an EBNA-2 deficient mutant virus. Surprisingly, and in contrast to previous reports, these viral mutants are attenuated in growth transformation assays but give rise to permanently growing EBNA-3A negative B cell lines which exhibit reduced proliferation rates and elevated levels of apoptosis. Expression profiles of EBNA-3A deficient LCLs are characterized by 129 down-regulated and 167 up-regulated genes, which are significantly enriched for genes involved in apoptotic processes or cell cycle progression like the tumor suppressor gene p16/INK4A, or might contribute to essential steps of the viral life cycle in the infected host. In addition, EBNA-3A cellular target genes remarkably overlap with previously identified targets of EBNA-2. This study comprises the first genome wide expression profiles of EBNA-3A target genes generated within the complex network of viral proteins of the growth transformed B cell and permits a more detailed understanding of EBNA-3A's function and contribution to viral pathogenesis. Epstein-Barr virus (EBV) infects primary human B cells and establishes a latent infection, which leads to permanently growing B cell cultures. These growth transformed B cells express a well defined set of latent viral genes, which are also expressed in post-transplant lymphomas of immunosuppressed patients. In a concerted action these latent viral proteins drive cellular proliferation and prevent apoptosis. For this study, recombinant Epstein-Barr virus mutants that lack the gene for the Epstein-Barr virus nuclear antigen-3A (EBNA-3A) were generated. EBNA-3A is a transcriptional modulator of gene expression. We show here that EBNA-3A deficient growth transformed B cells can be established in vitro. Our results suggest that EBNA-3A supports viability but is not absolutely essential for proliferation of the infected B cell. By virtue of the established EBNA-3A deficient cell lines, we could for the first time identify a broad array of cellular target genes controlled by EBNA-3A in EBV infected B cells. These EBNA-3A target genes will permit a more detailed understanding of EBNA-3A's function and contribution to viral pathogenesis.
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影响因子：
3.5
作者：
CHEREPANOV, PP;WACKERNAGEL, W
通讯作者：
WACKERNAGEL, W
影响因子：
8
作者：
Jeanes, A.;Gottardi, C. J.;Yap, A. S.
通讯作者：
Yap, A. S.
影响因子：
5.4
作者：
Bain, M;Watson, RJ;Allday, MJ
通讯作者：
Allday, MJ
DOI：
10.1111/j.2517-6161.1995.tb02031.x
发表时间：
1995-01-01
影响因子：
5.8
作者：
BENJAMINI, Y;HOCHBERG, Y
通讯作者：
HOCHBERG, Y
影响因子：
5.4
作者：
KEMPKES, B;PICH, D;HAMMERSCHMIDT, W
通讯作者：
HAMMERSCHMIDT, W