Inhibitions of human parainfluenza virus type 2 replication by ribavirin and mycophenolate mofetil are restored by guanosine and S-(4-nitrobenzyl)-6-thioinosine
Inhibitions of human parainfluenza virus type 2 replication by ribavirin and mycophenolate mofetil are restored by guanosine and S-(4-nitrobenzyl)-6-thioinosine
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DOI：
10.5582/ddt.2019.01084
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发表时间：
2019-12-01
影响因子：
3.1
通讯作者：
Komada, Hiroshi
中科院分区：
文献类型：
--
作者：
Uematsu, Jun;Sakai-Sugino, Kae;Komada, Hiroshi
研究了核苷类似物抗病毒药物（利巴韦林）和非核苷药物（吗替麦考酚酯）对人副流感病毒 2 型（hPIV-2）的抗病毒活性，还研究了鸟苷和 S-(4-硝基苄基)-6-硫代肌苷（NBTI：平衡核苷转运蛋白 1 抑制剂）抑制作用的恢复。利巴韦林 (RBV) 和吗替麦考酚酯 (MMF) 抑制 hPIV-2 诱导的细胞融合。 RBV 和 MMF 都显着减少了细胞释放的病毒数量。通过聚合酶链式反应 (PCR) 和实时 PCR 测定，病毒基因组合成受到 RBV 和 MMF 的抑制。 mRNA 合成也减少了。间接免疫荧光研究表明 RBV 和 MMF 在很大程度上抑制病毒蛋白合成。使用表达无基质蛋白的hPIV-2的重组绿色荧光蛋白（GFP）（rhPIV-2 Delta MGFP），发现病毒进入细胞和多核巨细胞形成几乎被RBV和MMF完全阻断。 RBV 和 MMF 不会破坏肌动蛋白微丝或微管。鸟苷和NBTI均完全或部分逆转RBV和MMF对病毒复制、基因组RNA、mRNA和蛋白质的合成以及多核巨细胞形成的抑制。 NBTI对肌动蛋白微丝造成少量损伤，但对微管没有影响。 RBV和MMF均抑制hPIV-2的复制，主要是通过抑制病毒基因组RNA、mRNA和蛋白质合成。鸟苷几乎完全恢复了抑制作用。这些结果表明抑制的主要机制是细胞内 GTP 库的耗尽。
The antiviral activities of a nucleoside analog antiviral drug (ribavirin) and a non-nucleoside drug (mycophenolate mofetil) against human parainfluenza virus type 2 (hPIV-2) were investigated, and the restoration of the inhibition by guanosine and S-(4-nitrobenzyl)-6-thioinosine (NBTI: equilibrative nucleoside transporter 1 inhibitor) were also investigated. Ribavirin (RBV) and mycophenolate mofetil (MMF) inhibited cell fusion induced by hPIV-2. Both RBV and MMF considerably reduced the number of viruses released from the cells. Virus genome synthesis was inhibited by RBV and MMF as determined by polymerase chain reaction (PCR) and real time PCR. mRNA syntheses were also reduced. An indirect immunofluorescence study showed that RBV and MMF largely inhibited viral protein syntheses. Using a recombinant green fluorescence protein (GFP)-expressing hPIV-2 without matrix protein (rhPIV-2 Delta MGFP), it was found that virus entry into the cells and multinucleated giant cell formation were almost completely blocked by RBV and MMF. RBV and MMF did not disrupt actin microfilaments or microtubules. Both guanosine and NBTI completely or partially reversed the inhibition by RBV and MMF in the viral replication, syntheses of genome RNA, mRNA and protein, and multinucleated giant cell formation. NBTI caused a little damage in actin microfilaments, but had no effect on microtubules. Both RBV and MMF inhibited the replication of hPIV-2, mainly by inhibiting viral genome RNA, mRNA and protein syntheses. The inhibition was almost completely recovered by guanosine. These results indicate that the major mechanism of the inhibition is the depletion of intracellular GTP pools.