Chemical Induction of Endogenous Retrovirus Particles from the Vero Cell Line of African Green Monkeys
Chemical Induction of Endogenous Retrovirus Particles from the Vero Cell Line of African Green Monkeys
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DOI：
10.1128/jvi.00147-11
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发表时间：
2011-07-01
影响因子：
5.4
通讯作者：
Khan, Arifa S.
中科院分区：
文献类型：
--
作者：
Ma, Hailun;Ma, Yunkun;Khan, Arifa S.
内源性逆转录病毒序列以高拷贝数存在于所有物种的基因组中，并可能以rna的形式表达；然而，大多数是有缺陷的病毒生产。虽然已经从旧大陆猴和新大陆猴中分离出病毒，但还没有从非洲绿猴（AGM）组织或细胞系中产生内源性逆转录病毒的报道。我们最近开发了一种通过化学诱导评估潜伏病毒存在的逐步方法（Khan等人，生物制品37:196-201,2009）。基于该策略，确定了在agm衍生的Vero细胞系中研究可诱导的内源性逆转录病毒存在的最佳条件。5-氮杂胞苷（AzaC）和5'-碘-2'-脱氧尿苷（IUdR）产生低水平的逆转录酶活性；丁酸钠无检测。从过滤后的上清液超离心颗粒制备的rna gag、pol和env区扩增片段的核苷酸序列分析表明，AzaC可以诱导与猿猴内源性D型β -逆转录病毒（SERV）序列和狒狒内源性C型γ -逆转录病毒（BaEV）序列相关的内源性逆转录病毒颗粒，IUdR也可以诱导SERV序列。此外，SERV-和baev相关颗粒的rna序列存在异质性。经药物处理的AGM Vero细胞的感染性分析显示，在已知对D型猿猴逆转录病毒（srv）和BaEV敏感的细胞系中没有病毒复制。结果表明，AGM基因组中存在多个可诱导的内源性逆转录病毒基因座，这些基因座可以编码非传染性病毒样颗粒。
Endogenous retroviral sequences are present in high copy numbers in the genomes of all species and may be expressed as RNAs; however, the majority are defective for virus production. Although virus has been isolated from various Old World monkey and New World monkey species, there has been no report of endogenous retroviruses produced from African green monkey (AGM) tissues or cell lines. We have recently developed a stepwise approach for evaluating the presence of latent viruses by chemical induction (Khan et al., Biologicals 37: 196-201, 2009). Based upon this strategy, optimum conditions were determined for investigating the presence of inducible, endogenous retroviruses in the AGM-derived Vero cell line. Low-level reverse transcriptase activity was produced with 5-azacytidine (AzaC) and with 5'-iodo-2'-deoxyuridine (IUdR); none was detected with sodium butyrate. Nucleotide sequence analysis of PCR-amplified fragments from the gag, pol, and env regions of RNAs, prepared from ultracentrifuged pellets of filtered supernatants, indicated that endogenous retrovirus particles related to simian endogenous type D betaretrovirus (SERV) sequences and baboon endogenous virus type C gammaretrovirus (BaEV) sequences were induced by AzaC, whereas SERV sequences were also induced by IUdR. Additionally, sequence heterogeneity was seen in the RNAs of SERV- and BaEV-related particles. Infectivity analysis of drug-treated AGM Vero cells showed no virus replication in cell lines known to be susceptible to type D simian retroviruses (SRVs) and to BaEV. The results indicated that multiple, inducible endogenous retrovirus loci are present in the AGM genome that can encode noninfectious, viruslike particles.