DETECTION OF HERPES-SIMPLEX VIRUS TYPE-1 LATENCY ASSOCIATED TRANSCRIPTS IN CORNEAL CELLS OF INBRED MICE BY INSITU HYBRIDIZATION
DETECTION OF HERPES-SIMPLEX VIRUS TYPE-1 LATENCY ASSOCIATED TRANSCRIPTS IN CORNEAL CELLS OF INBRED MICE BY INSITU HYBRIDIZATION
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DOI：
10.1097/00003226-199209000-00013
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发表时间：
1992-09-01
期刊：
影响因子：
2.8
通讯作者：
PETRASH, JM
中科院分区：
文献类型：
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作者：
ABGHARI, SZ;STULTING, RD;PETRASH, JM
我们先前已恢复单纯疱疹病毒1型（HSV）从角膜潜伏感染的小鼠在体外培养。然而，可以认为，这些数据不能明确区分持续性和潜伏性角膜感染。我们现在已经使用RNA原位杂交来解决这个问题，通过确定HSV基因在潜伏感染期间的BALB/c小鼠角膜中的表达。在用HSV局部角膜接种后2至4个月，当不存在活动性眼部疾病或感染性病毒时，取出角膜并用胶原酶消化。将从两个角膜汇集的解离细胞与H-3-或S-35-标记的2.6-kb单链RNA探针杂交，以检测有义和反义ICP-0转录物。25%的池与反义ICP-0（潜伏相关转录物，LAT）的探针杂交，而仅3%与ICP-0的探针杂交（p < 0.03）。在阳性池中的细胞中，0.6-7.0%显示LAT阳性杂交信号。通过培养来自探针池或对照潜伏感染角膜的上清液，未发现感染性病毒。这些数据提供了进一步的证据，HSV可以在小鼠角膜中建立真正的潜伏感染。
We have previously recovered herpes simplex virus type 1 (HSV) from the corneas of latently infected mice by cultivation in vitro. It could be argued, however, that these data do not definitively distinguish between persistent and latent corneal infection. We have now used RNA hybridization in situ to resolve this question by determining the expression of HSV genes in the corneas of BALB/c mice during latent infection. Two to four months after topical corneal inoculation with HSV, when no active ocular disease or infectious virus was present, corneas were removed and digested with collagenase. Dissociated cells pooled from two corneas were hybridized with H-3- or S-35-labeled 2.6-kb single-stranded RNA probes to detect sense and antisense ICP-0 transcripts. Twenty-five percent of the pools hybridized with the probe for antisense ICP-0 (latency-associated transcript, LAT), while only 3% hybridized with the probe for ICP-0 (p < 0.03). Of the cells in positive pools, 0.6-7.0% showed a positive hybridization signal for LAT. No infectious virus was found by culture of supernatants from the probed pools or control latently infected corneas. These data provide further evidence that HSV can establish a true latent infection in the mouse cornea.