title: HSV-1 migration in latently infected and naive rabbits after penetrating keratoplasty.
abstract: OBJECTIVE
  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.
authors.affiliation: Department of Ophthalmology, Lions Eye Research Laboratories, LSU Eye Center, Louisiana State University Health Sciences, Center School of Medicine, New Orleans, USA.
authors.forename: X
authors.initials: X
authors.lastname: Zheng
authors.forename: M E
authors.initials: ME
authors.lastname: Marquart
authors.forename: J M
authors.initials: JM
authors.lastname: Loustch
authors.forename: P
authors.initials: P
authors.lastname: Shah
authors.forename: B
authors.initials: B
authors.lastname: Sainz
authors.forename: A
authors.initials: A
authors.lastname: Ray
authors.forename: R J
authors.initials: RJ
authors.lastname: O'Callaghan
authors.forename: H E
authors.initials: HE
authors.lastname: Kaufman
authors.forename: J M
authors.initials: JM
authors.lastname: Hill
mesh_terms: D000818:Animals; D003315:Cornea / Q000294:innervation / Q000821:virology*; D017931:DNA Primers / Q000737:chemistry; D004279:DNA, Viral / Q000032:analysis; D004837:Epinephrine / Q000494:pharmacology; D015870:Gene Expression / Q000235:genetics; D005814:Genes, Viral / Q000235:genetics; D006085:Graft Survival; D018259:Herpesvirus 1, Human / Q000235:genetics / Q000502:physiology*; D007478:Iontophoresis; D016849:Keratitis, Herpetic / Q000473:pathology / Q000821:virology*; D015948:Keratoplasty, Penetrating; D016133:Polymerase Chain Reaction; D011817:Rabbits; D013666:Tears / Q000821:virology; D012668:Trigeminal Ganglion / Q000821:virology; D014775:Virus Activation / Q000187:drug effects; D017735:Virus Latency / Q000502:physiology*; D017201:Virus Shedding / Q000502:physiology
chemical_list: D017931:DNA Primers; D004279:DNA, Viral; D004837:Epinephrine