THYMIDINE KINASE-NEGATIVE HERPES-SIMPLEX VIRUS MUTANTS ESTABLISH LATENCY IN MOUSE TRIGEMINAL GANGLIA BUT DO NOT REACTIVATE
THYMIDINE KINASE-NEGATIVE HERPES-SIMPLEX VIRUS MUTANTS ESTABLISH LATENCY IN MOUSE TRIGEMINAL GANGLIA BUT DO NOT REACTIVATE
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DOI：
10.1073/pnas.86.12.4736
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发表时间：
1989-06-01
影响因子：
11.1
通讯作者：
KNIPE, DM
中科院分区：
文献类型：
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作者：
COEN, DM;KOSZVNENCHAK, M;KNIPE, DM
哺乳动物宿主的单纯疱疹病毒感染涉及在原发部位（例如角膜）的裂解性复制、通过轴突运输易位至感觉神经节和复制，以及在继发部位（神经节神经元）的潜伏感染。病毒编码的胸苷激酶是阿昔洛韦等抗病毒药物的靶标，它对于裂解性复制不是必需的，但显然是次级复制位点和潜伏感染的某些阶段所必需的。为了确定病毒发病机制中需要该酶的特定阶段，我们构建了具有阿昔洛韦抗性且未表现出可检测到的胸苷激酶活性的病毒缺失突变体。接种小鼠角膜后，突变体在眼睛中复制到高滴度，但在三叉神经节中的急性复制严重受损，并且在与许可细胞共培养时未能从神经节中重新激活。然而，潜伏期相关的转录物在突变体感染小鼠的神经节神经元核中表达，并且用第二种病毒对神经节的双重感染拯救了潜伏的突变体病毒。因此，与广泛接受的假设相反，胸苷激酶阴性突变体建立了潜伏感染，这意味着胸苷激酶活性和神经节复制都不是潜伏建立所必需的。相反，胸苷激酶似乎是从潜伏期重新激活所必需的。这些结果表明，阿昔洛韦耐药病毒可以在临床环境中建立潜伏感染，并对使用基因工程疱疹病毒将外源基因传递到神经元具有影响。
Herpes simplex virus infection of mammalian hosts involves lytic replication at a primary site, such as the cornea, translocation by axonal transport to sensory ganglion and replication, and latent infection at a secondary site, ganglionic neurons. The virus-encoded thymidine kinase, which is a target for antiviral drugs such as acyclovir, is not essential for lytic replication yet evidently is required at the secondary site for replication and some phase of latent infection. To determine the specific stage in viral pathogenesis at which this enzyme is required, we constructed virus deletion mutants that were acyclovir resistant and exhibited no detectable thymidine kinase activity. After corneal inoculation of mice, the mutants replicated to high titers in the eye but were severely impaired for acute replication in trigeminal ganglia and failed to reactivate from ganglia upon cocultivation with permissive cells. Nevertheless, latency-associated transcripts were expressed in neuronal nuclei of ganglia from mutant-infected mice and superinfection of the ganglia with a second virus rescued the latent mutant virus. Thus, contrary to a widely accepted hypothesis, the thymidine kinase-negative mutants established latent infections, implying that neither thymidine kinase activity nor ganglionic replication is neccessary for establishment of latency. Rather, thymidine kinase appears to be necessary for reactivation from latency. These results suggest that acyclovir-resistant viruses could establish latent infections in clinical settings and have implications for the use of genetically engineered herpesviruses to deliver foreign genes to neurons.