Department of Microbiology and Immunology, Cornell University, Ithaca, New York, USA College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, China.
Department of Microbiology and Immunology, Cornell University, Ithaca, New York, USA.
J Virol. 2014 Oct;88(19):11121-9. doi: 10.1128/JVI.01540-14. Epub 2014 Jul 9.
Previous reports showed that raltegravir, a recently approved antiviral compound that targets HIV integrase, can inhibit the nuclease function of human cytomegalovirus (HCMV terminase) in vitro. In this study, subtoxic levels of raltegravir were shown to inhibit the replication of four different herpesviruses, herpes simplex virus 1 (HSV-1), HSV-2, HCMV, and mouse cytomegalovirus, by 30- to 700-fold, depending on the dose and the virus tested. Southern blotting and quantitative PCR revealed that raltegravir inhibits DNA replication of HSV-1 rather than cleavage of viral DNA. A raltegravir-resistant HSV-1 mutant was generated by repeated passage in the presence of 200 μM raltegravir. The genomic sequence of the resistant virus, designated clone 7, contained mutations in 16 open reading frames. Of these, the mutations F198S in unique long region 15 (UL15; encoding the large terminase subunit), A374V in UL32 (required for DNA cleavage and packaging), V296I in UL42 (encoding the DNA polymerase accessory factor), and A224S in UL54 (encoding ICP27, an important transcriptional regulator) were introduced independently into the wild-type HSV-1(F) genome, and the recombinant viruses were tested for raltegravir resistance. Viruses bearing both the UL15 and UL32 mutations inserted within the genome of the UL42 mutant were also tested. While the UL15, UL32, and UL54 mutant viruses were fully susceptible to raltegravir, any virus bearing the UL42 mutation was as resistant to raltegravir as clone 7. Overall, these results suggest that raltegravir may be a valuable therapeutic agent against herpesviruses and the antiviral activity targets the DNA polymerase accessory factor rather than the nuclease activity of the terminase.
This paper shows that raltegravir, the antiretrovirus drug targeting integrase, is effective against various herpesviruses. Drug resistance mapped to the herpesvirus DNA polymerase accessory factor, which was an unexpected finding.
先前的报告表明，拉替拉韦是一种最近被批准的靶向 HIV 整合酶的抗病毒化合物，能够在体外抑制人巨细胞病毒（HCMV 末端酶）的核酸酶功能。在这项研究中，亚毒性水平的拉替拉韦能够抑制四种不同的疱疹病毒，单纯疱疹病毒 1（HSV-1）、HSV-2、HCMV 和鼠巨细胞病毒的复制，抑制效果取决于剂量和测试的病毒，范围在 30 到 700 倍之间。Southern 印迹和定量 PCR 显示，拉替拉韦抑制 HSV-1 的 DNA 复制而不是切割病毒 DNA。通过在 200 μM 拉替拉韦存在下反复传代，生成了对拉替拉韦具有抗性的 HSV-1 突变体。将具有抗性的病毒命名为克隆 7，其基因组序列包含在 16 个开放阅读框中的突变。其中，独特长区 15（UL15；编码大末端酶亚基）中的 F198S、UL32 中的 A374V（需要 DNA 切割和包装）、UL42 中的 V296I（编码 DNA 聚合酶辅助因子）和 UL54 中的 A224S（编码 ICP27，一种重要的转录调节剂）的突变是独立引入到野生型 HSV-1(F)基因组中的，并且对重组病毒进行了拉替拉韦抗性测试。还测试了携带 UL15 和 UL32 突变的病毒在 UL42 突变病毒基因组中的插入情况。虽然 UL15、UL32 和 UL54 突变病毒对拉替拉韦完全敏感，但任何携带 UL42 突变的病毒都与克隆 7 一样对拉替拉韦具有抗性。总体而言，这些结果表明，拉替拉韦可能是一种针对疱疹病毒的有价值的治疗药物，抗病毒活性靶向 DNA 聚合酶辅助因子而不是末端酶的核酸酶活性。
本文表明，针对整合酶的抗逆转录病毒药物拉替拉韦对各种疱疹病毒有效。耐药性映射到疱疹病毒 DNA 聚合酶辅助因子，这是一个意外的发现。
