Establishment of a quiescent herpes simplex virus type 1 infection in neurally-differentiated PC12 cells
Establishment of a quiescent herpes simplex virus type 1 infection in neurally-differentiated PC12 cells
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DOI：
10.3109/13550289909015812
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发表时间：
1999-06-01
影响因子：
3.2
通讯作者：
Miller, CS
中科院分区：
文献类型：
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作者：
Danaher, RJ;Jacob, RJ;Miller, CS
用神经生长因子（Nd-PC 12）分化的大鼠嗜铬细胞瘤（PC 12）细胞用于研究可逆的非生产性单纯疱疹病毒1型（HSV-1）感染的建立。结果表明：（1）Nd-PC_（12）培养物可长期保存（>7周）仅当在不存在血清的情况下铺在胶原包被的培养皿上时的非分裂培养物;（ii）在短暂存在无环鸟苷（ACV）的情况下用HSV-1株科斯和17感染Nd-PC 12在去除ACV时，导致所有培养物均无可检测水平的感染性病毒，并且在随后的8周内不需要ACV来维持非生产性静止状态;（iii）这些持续感染和静止（QIF）-PC 12培养物显示在ACV停药后的前2周内分别以低频率（5%）和高频率（92 - 100%）自发和毛喉素诱导的病毒产生。(iv)与其他体外模型相反，HSV-1在去除神经生长因子后未能重新激活。(v)高百分比的QIF-PC 12培养物（50-100%）产生病毒，以响应毛喉素治疗长达7周后ACV停药。(vi)HSV-1产生基因（即α 0、α 4、α 27、U-L，30和U-L，18）的表达在ACV存在下急剧下降，并且在ACV去除后保持不可检测或继续下降，而LAT和宿主基因G3 PDH的水平在整个31天的研究期间保持相对恒定，如通过RT-PCR测量的。这些结果表明，QIF-PC 12细胞提供了一种新的神经元细胞培养系统，可以增强我们在没有复制抑制剂的情况下研究HSV-1从隐蔽的、潜伏样的、非生产性状态再活化的能力。
Rat pheochromocytoma (PC12) cells differentiated with nerve growth factor (Nd-PC12) were used to investigate the establishment of a non-productive herpes simplex virus type 1 (HSV-1) infection that is reversible. The results of this work are as follows: (i) Nd-PC12 cultures could be maintained as long term (>7 weeks) non-dividing cultures only when plated on collagen-coated dishes in the absence of serum; (ii) Infection of Nd-PC12 with HSV-1 strains KOS and 17 in the transient presence of acycloguanosine (ACV) resulted in all cultures free of detectable levels of infectious virus at the time of ACV removal and ACV was not needed to maintain the non-productive quiescent state in the subsequent 8 weeks; (iii) These persistently infected and quiescent (QIF)-PC12 cultures demonstrated both spontaneous and forskolin-inducible virus production, at low (5%) and high frequencies (92 - 100%), respectively during the first 2 weeks post-ACV withdrawal. (iv) In contrast to other in vitro models, HSV-1 failed to reactivate following removal of nerve growth factor. (v) A high percentage of QIF-PC12 cultures (50-100%) produced virus in response to forskolin treatment as long as 7 weeks post-ACV withdrawal. (vi) Expression of HSV-1 productive genes (i.e. alpha 0, alpha 4, alpha 27, U-L,30 and U-L,18) dropped precipitously in the presence of ACV and remained undetectable or continued to decline following its removal, whereas the levels of LAT and the host gene G3PDH remained relatively constant throughout the 31 day study period as measured by RT-PCR. These results indicate that QIF-PC12 cells offer a novel, neuronal cell culture system that may enhance our ability to study HSV-1 reactivation from a cryptic, latent-like, non-productive state in the absence of replication inhibitors.