Ecob-Prince M S, Hassan K, Denheen M T, Preston C M
University of Glasgow Department of Neurology, Southern General Hospital, UK.
J Gen Virol. 1995 Jun;76 ( Pt 6):1527-32. doi: 10.1099/0022-1317-76-6-1527.
Explanation into culture of dorsal root ganglia (DRG) latently infected with herpes simplex virus type 1 (HSV-1) causes reactivation of the virus. Previous studies have suggested that either latency-associated transcripts (LATs) were removed as an early consequence of reactivation or, alternatively, there was a population of latently infected cells which did not contain LATs. We have now attempted to detect this population of neurons by inserting a reporter gene (Escherichia coli lacZ gene), under the control of promoters other than LAT, into the HSV-1 strain 17 mutant in 1814, which was used in the earlier studies. One of these promoters, the human cytomegalovirus enhancer, resulted in weak expression of beta-galactosidase in DRG neurons for at least 5 months. The pattern of staining was predominantly homogeneous in neurons at 3 or 5 days post-infection or at 3 days post-explanation, and was predominantly speckled in latently infected neurons (1 to 5 months post-infection). About 30% of the beta-galactosidase-positive neurons did not contain LATs by in situ hybridization. However, the detergents used to enable penetration of the substrate for beta-galactosidase had also reduced the levels of the LATs; in neurons which originally had only small numbers of LATs this may have reduced levels to below those detectable by the methods used. There was, therefore, no unequivocal evidence for a population of latently HSV-1-infected cells which did not express LATs.
对潜伏感染1型单纯疱疹病毒(HSV - 1)的背根神经节(DRG)进行传代培养会导致病毒重新激活。先前的研究表明,要么潜伏相关转录本(LATs)在重新激活的早期就被清除,要么存在一群不含有LATs的潜伏感染细胞。我们现在试图通过将一个在LAT以外的启动子控制下的报告基因(大肠杆菌lacZ基因)插入到早期研究中使用的HSV - 1 17株1814突变体中来检测这群神经元。其中一个启动子,即人巨细胞病毒增强子,导致DRG神经元中β - 半乳糖苷酶至少5个月的弱表达。在感染后3天或5天或传代后3天,神经元中的染色模式主要是均匀的,而在潜伏感染的神经元中(感染后1至5个月)主要是斑点状的。通过原位杂交,约30%的β - 半乳糖苷酶阳性神经元不含有LATs。然而,用于使β - 半乳糖苷酶底物渗透的去污剂也降低了LATs的水平;在原本只有少量LATs的神经元中,这可能使水平降低到低于所用方法可检测的水平。因此,没有明确的证据表明存在一群不表达LATs的潜伏感染HSV - 1的细胞。