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(Investigative Ophthalmology and Visual Science. 1999;40:2490-2497.)
© 1999 by The Association for Research in Vision and Ophthalmology, Inc.

HSV-1 Migration in Latently Infected and Naive Rabbits after Penetrating Keratoplasty

Xiaodong Zheng, Mary E. Marquart, Jeannette M. Loustch, Pulin Shah, Bruno Sainz, Arnab Ray, Richard J. O’Callaghan, Herbert E. Kaufman and James M. Hill

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

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: 17{Delta}Pst, a latency associated transcript (LAT) negative, low-reactivating virus and 17Pr, a high-reactivating, LAT-positive rescuant of 17{Delta}Pst. 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 17{Delta}Pst (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 17{Delta}Pst (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 17{Delta}Pst has a significantly reduced ability to replicate and migrate.




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