Effects of ribavirin on the replication and genetic stability of porcine reproductive and respiratory syndrome virus.
Effects of ribavirin on the replication and genetic stability of porcine reproductive and respiratory syndrome virus.
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DOI：
10.1186/s12917-015-0330-z
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发表时间：
2015-02-07
影响因子：
2.6
通讯作者：
Kim WI
中科院分区：
文献类型：
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作者：
Khatun A;Shabir N;Yoon KJ;Kim WI
尽管修饰活病毒（MLV）疫苗通常用于控制猪生殖与呼吸综合征病毒（PRRSV），但由于MLV在猪体内复制期间迅速恢复毒力，因此存在安全问题。以往的研究表明，突变病毒产生的致死性诱变驱动的抗病毒诱变剂，这些病毒具有更高的遗传稳定性相比，其亲本株，因为他们获得了抗随机突变。因此，在本研究中探索了这种策略以稳定PRRSV基因组。评价了4种抗病毒诱变剂（利巴韦林、5-氟尿嘧啶、5-氮杂胞苷和阿米洛利）对2型PRRSV原型VR 2332的抗病毒效果。在诱变剂中，利巴韦林和5-氟尿嘧啶对VR 2332有显著的抗病毒作用。因此，VR 2332在MARC-145细胞中在存在几种浓度的利巴韦林的情况下连续传代，以促进利巴韦林抗性突变体的出现。两个利巴韦林耐药突变体，RVRp 13和RVRp 22，出现从连续传代中存在的0.1和0.2 mM利巴韦林，分别。在MARC-145细胞中评价了这些抗性突变体的遗传稳定性，并与VR 2332进行了比较。正如预期的那样，利巴韦林耐药突变体与其亲本病毒相比表现出更高的遗传稳定性。总之，利巴韦林和5-氟尿嘧啶有效地抑制了MARC-145细胞中的PRRSV复制。然而，当用低浓度（≤0.2 mM）利巴韦林处理时，出现了利巴韦林耐药突变体，并且这些突变体在细胞培养物中连续传代期间在遗传上更稳定。
Although modified live virus (MLV) vaccines are commonly used for porcine reproductive and respiratory syndrome virus (PRRSV) control, there have been safety concerns due to the quick reversion of MLV to virulence during replication in pigs. Previous studies have demonstrated that mutant viruses emerged from lethal mutagenesis driven by antiviral mutagens and that those viruses had higher genetic stability compared to their parental strains because they acquired resistance to random mutation. Thus, this strategy was explored to stabilize the PRRSV genome in the current study. Four antiviral mutagens (ribavirin, 5-fluorouracil, 5-azacytidine, and amiloride) were evaluated for their antiviral effects against VR2332, a prototype of type 2 PRRSV. Among the mutagens, ribavirin and 5-fluorouracil had significant antiviral effects against VR2332. Consequently, VR2332 was serially passaged in MARC-145 cells in the presence of ribavirin at several concentrations to facilitate the emergence of ribavirin-resistant mutants. Two ribavirin-resistant mutants, RVRp13 and RVRp22, emerged from serial passages in the presence of 0.1 and 0.2 mM ribavirin, respectively. The genetic stability of these resistant mutants was evaluated in MARC-145 cells and compared with VR2332. As expected, the ribavirin-resistant mutants exhibited higher genetic stability compared to their parental virus. In summary, ribavirin and 5-fluorouracil effectively suppressed PRRSV replication in MARC-145 cells. However, ribavirin-resistant mutants emerged when treated with low concentrations (≤0.2 mM) of ribavirin, and those mutants were genetically more stable during serial passages in cell culture.