穿透性角膜移植术后单纯疱疹病毒1型在潜伏感染和未感染家兔中的迁移情况

HSV-1 migration in latently infected and naive rabbits after penetrating keratoplasty.

作者信息

Zheng X, Marquart M E, Loustch J M, Shah P, Sainz B, Ray A, O'Callaghan R J, Kaufman H E, Hill J M

机构信息

Department of Ophthalmology, Lions Eye Research Laboratories, LSU Eye Center, Louisiana State University Health Sciences, Center School of Medicine, New Orleans, USA.

出版信息

Invest Ophthalmol Vis Sci. 1999 Oct;40(11):2490-7.

PMID:10509641

Abstract

PURPOSE

To investigate the migration of herpes simplex virus type 1 (HSV-1) between latently infected and naive corneal tissues and trigeminal ganglion (TG) in rabbits after penetrating keratoplasty (PKP) and transcorneal epinephrine iontophoresis.

METHODS

Two mutants, genetically constructed from HSV-1 strain 17syn+, were used to inoculate rabbit corneas: 17deltaPst, a latency associated transcript (LAT) negative, low-reactivating virus and 17Pr, a high-reactivating, LAT-positive rescuant of 17deltaPst. Latently infected rabbits were given corneal allografts from naive rabbits, and naive rabbits received grafts from latently infected rabbits. Ninety days after PKP, groups of the transplanted rabbits were induced to reactivate by transcorneal epinephrine iontophoresis, but others were not induced. Viral shedding was monitored by tear film cultures. Rabbits were killed 5 days after iontophoresis. Transplanted grafts, recipient corneal rims, and corresponding TG were obtained. Nucleic acids were extracted and amplified for detection of HSV-1 DNA and viral gene transcription.

RESULTS

In naive rabbits receiving grafts transplanted from rabbits latently infected with 17Pr (LAT+), 3 of 6 corneal rims contained HSV DNA after induction. In contrast, none of the 5 corneal rims from naive rabbits receiving grafts from rabbits latent with 17deltaPst (LAT-) contained viral DNA. Viral DNA and gene transcripts were detected in 2 of 6 TG from naive rabbits that received grafts from 17Pr (LAT+) latently infected rabbits. In recipient corneal rims and TG of latently infected rabbits receiving grafts from naive rabbits, viral DNA concentration was significantly greater with induced reactivation, compared with the results in noninduced rabbits. The amount of viral DNA in naive grafts transplanted into 17Pr (LAT+) latently infected rabbits was significantly higher with induction than without induction (P = 0.018). More viral DNA and viral gene transcripts were found in tissues from rabbits latently infected with 17Pr (LAT+) than in rabbits latently infected with 17deltaPst (LAT-).

CONCLUSIONS

Corneas from latently infected rabbits contain HSV-1 DNA that can replicate after induced reactivation. Viral migration can occur in both anterograde and retrograde directions between the transplanted graft and the recipient corneal rim and TG. The LAT negative HSV-1 construct 17deltaPst has a significantly reduced ability to replicate and migrate.

摘要

目的

研究穿透性角膜移植术（PKP）和经角膜肾上腺素离子导入后，单纯疱疹病毒1型（HSV-1）在兔潜伏感染和未感染的角膜组织与三叉神经节（TG）之间的迁移情况。

方法

使用从HSV-1 17syn+株基因构建的两种突变体接种兔角膜：17deltaPst，一种潜伏期相关转录物（LAT）阴性、低再激活病毒；17Pr，17deltaPst的高再激活、LAT阳性拯救病毒。将潜伏感染的兔给予来自未感染兔的角膜同种异体移植，未感染的兔接受来自潜伏感染兔的移植物。PKP术后90天，对移植兔分组进行经角膜肾上腺素离子导入诱导再激活，其他组不进行诱导。通过泪膜培养监测病毒脱落情况。离子导入后5天处死兔。获取移植的移植物、受体角膜边缘和相应的TG。提取核酸并进行扩增以检测HSV-1 DNA和病毒基因转录。

结果

在接受从潜伏感染17Pr（LAT+）的兔移植的移植物的未感染兔中，诱导后6个角膜边缘中有3个含有HSV DNA。相比之下，接受从潜伏感染17deltaPst（LAT-）的兔移植的移植物的未感染兔的5个角膜边缘中均未检测到病毒DNA。在接受从潜伏感染17Pr（LAT+）的兔移植的移植物的未感染兔的6个TG中，有2个检测到病毒DNA和基因转录物。在接受来自未感染兔的移植物的潜伏感染兔的受体角膜边缘和TG中，与未诱导的兔相比，诱导再激活后病毒DNA浓度显著更高。移植到潜伏感染17Pr（LAT+）的兔中的未感染移植物中，诱导组的病毒DNA量显著高于未诱导组（P = 0.018）。与潜伏感染17deltaPst（LAT-）的兔相比，潜伏感染17Pr（LAT+）的兔的组织中发现更多的病毒DNA和病毒基因转录物。