Inhibition of dengue virus serotypes 1 to 4 in Vero cell cultures with morpholino oligomers
Inhibition of dengue virus serotypes 1 to 4 in Vero cell cultures with morpholino oligomers
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DOI：
10.1128/jvi.79.8.5116-5128.2005
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发表时间：
2005-04-01
影响因子：
5.4
通讯作者：
Stein, DA
中科院分区：
文献类型：
--
作者：
Kinney, RM;Huang, CYH;Stein, DA
评价了5种登革病毒特异性R5 F2 R4肽缀合的磷酰二胺吗啉代寡聚物（P4-PMO）在哺乳动物细胞培养物中抑制登革病毒血清型2（DEN-2病毒）复制的能力。DEN-2病毒16681的初始生长曲线在与20 μ M P4-PMO化合物温育的Vero细胞中获得。在感染后6天，靶向DEN-2病毒基因组3 '末端核苷酸的P4-PMO和随机序列P4-PMO显示出对DEN-2病毒滴度的相对较小的抑制（分别为0.1和0.9 log（10））。靶向单个开放阅读框的AUG翻译起始位点区域和5'环化序列区域的P4-PMO具有中等活性，产生1.6-和1.8-log（10）减少。两种P4-PMO化合物，5 'SL和3' CS（分别靶向5 '-末端核苷酸和3'环化序列区域）是高度有效的，在用DEN-2病毒感染后6天，与对照相比，每种化合物都将病毒滴度降低大于5.7 log（10）。进一步的实验表明，5 'SL和3' CS抑制DEN-2病毒复制的作用具有剂量依赖性和序列特异性。用10 μ M 3 ′ CS处理降低了所有四种DEN病毒血清型的滴度，即，DEN-1（菌株16007）、DEN-2（16681）、DEN-3（16562）和DEN-4（1036）病毒超过4 log（10），在大多数情况下低于检测限。3 'CS功效的程度受与细胞的病毒感染相关的化合物应用的时机的影响。5 'SL和3' CS P4-PMO不抑制西尼罗病毒NY 99在Vero细胞中的复制。这些数据表明，有必要进一步评价5 ′ SL和3 ′ CS化合物作为潜在的DEN病毒治疗剂。
Five dengue (DEN) virus-specific R5F2R4 peptide-conjugated phosphorodiamidate morpholino oligomers (P4-PMOs) were evaluated for their ability to inhibit replication of DEN virus serotype 2 (DEN-2 virus) in mammalian cell culture. Initial growth curves of DEN-2 virus 16681 were obtained in Vero cells incubated with 20 mu M P4-PMO compounds. At 6 days after infection, a P4-PMO targeting the 3'-terminal nucleotides of the DEN-2 virus genome and a random-sequence P4-PMO showed relatively little suppression of DEN-2 virus titer (0.1 and 0.9 log(10), respectively). P4-PMOs targeting the AUG translation start site region of the single open reading frame and the 5' cyclization sequence region had moderate activity, generating 1.6- and 1.8-log(10) reductions. Two P4-PMO compounds, 5'SL and 3'CS (targeting the 5'-terminal nucleotides and the 3' cyclization sequence region, respectively), were highly efficacious, each reducing the viral titer by greater than 5.7 log(10) compared to controls at 6 days after infection with DEN-2 virus. Further experiments showed that 5'SL and 3'CS inhibited DEN-2 virus replication in a dose-dependent and sequence-specific manner. Treatment with 10 mu M 3'CS reduced the titers of all four DEN virus serotypes, i.e., DEN-1 (strain 16007), DEN-2 (16681), DEN-3 (16562), and DEN-4 (1036) viruses by over 4 log(10), in most cases to below detectable limits. The extent of 3'CS efficacy was affected by the timing of compound application in relation to viral infection of the cells. The 5'SL and 3'CS P4-PMOs did not suppress the replication of West Nile virus NY99 in Vero cells. These data indicate that further evaluation of the 5'SL and 3'CS compounds as potential DEN virus therapeutics is warranted.