Features Distinguishing Epstein-Barr Virus Infections of Epithelial Cells and B Cells: Viral Genome Expression, Genome Maintenance, and Genome Amplification
Features Distinguishing Epstein-Barr Virus Infections of Epithelial Cells and B Cells: Viral Genome Expression, Genome Maintenance, and Genome Amplification
复制标题
DOI：
10.1128/jvi.00108-09
复制
发表时间：
2009-08-01
影响因子：
5.4
通讯作者：
Rowe, Martin
中科院分区：
文献类型：
--
作者：
Shannon-Lowe, Claire;Adland, Emily;Rowe, Martin
EB病毒（EBV）与淋巴和上皮细胞起源的恶性疾病有关。EBV的嗜性是由于病毒的主要受体分子CD 21的B细胞限制性表达。然而，CD 21（-）上皮细胞的有效感染可以通过从EBV包被的B细胞转移来实现。我们在这里比较和对比的早期事件后，在体外感染的原代B细胞和上皮细胞。使用敏感的，定量的逆转录-PCR检测几个潜在的和裂解的成绩单和双色免疫荧光染色分析在单细胞水平上的表达，我们证实和扩展以前的报告表明，这两种细胞类型支持不同的转录模式。此外，尽管用一个或两个拷贝的EBV感染B细胞导致病毒基因组快速扩增至每个细胞> 20个拷贝，但在感染原代上皮细胞或未分化上皮细胞系后通常观察不到这种扩增。在上皮细胞中，EBNA 1表达仅在癌细胞中检测到。40%的EBER+细胞，并且EBV基因组随后在长期培养期间丢失。一个例外是AGS（胃癌细胞系）的感染导致EBNA 1表达的维持和EBV附加体的扩增。相反，B细胞，其中EBV附加体的扩增发生，甚至与复制缺陷型BZLF 1敲除病毒，扩增在AGS细胞依赖于早期裂解周期基因表达。这些数据突出了宿主细胞在基因组表达、扩增和维持方面对EBV感染结果的影响。
Epstein-Barr virus (EBV) is associated with malignant diseases of lymphoid and epithelial cell origin. The tropism of EBV is due to B-cell-restricted expression of CD21, the major receptor molecule for the virus. However, efficient infection of CD21(-) epithelial cells can be achieved via transfer from EBV-coated B cells. We compare and contrast here the early events following in vitro infection of primary B cells and epithelial cells. Using sensitive, quantitative reverse transcription-PCR assays for several latent and lytic transcripts and two-color immunofluorescence staining to analyze expression at the single cell level, we confirmed and extended previous reports indicating that the two cell types support different patterns of transcription. Furthermore, whereas infection of B cells with one or two copies of EBV resulted in rapid amplification of the viral genome to > 20 copies per cell, such amplification was not normally observed after infection of primary epithelial cells or undifferentiated epithelial lines. In epithelial cells, EBNA1 expression was detected in only ca. 40% of EBER+ cells, and the EBV genome was subsequently lost during prolonged culture. One exception was that infection of AGS, a gastric carcinoma line, resulted in maintenance of EBNA1 expression and amplification of the EBV episome. In contrast to B cells, where amplification of the EBV episome occurred even with a replication-defective BZLF1-knockout virus, amplification in AGS cells was dependent upon early lytic cycle gene expression. These data highlight the influence of the host cell on the outcome of EBV infection with regard to genome expression, amplification, and maintenance.