PURPOSE: The aim of the present study was to identify the genes that express in the nervous system when herpes simplex virus type 1 (HSV-1) reactivates from latency by using subtraction cloning following epinephrine iontophoresis. METHODS: The corneas of New Zealand White rabbits (2 to 2.5 kg) were topically inoculated with 5 x 10(5) PFU of plaque-purified HSV-1 strain McKrae. Corneal infection was monitored by slit lamp examination (SLE) on days 3-7 after infection. At 30 d post-infection, eyes were assessed for infectious virus by ocular swabs. Disappearance of virus in ocular swabs signified latency for HSV-1. Latently-infected rabbits were subjected to transcorneal iontophoresis. The messenger RNA (mRNA) was isolated from the trigeminal ganglia and used for subtraction cloning and subtraction library synthesis. RESULTS: Our results indicate 18 genes with increased expression and 6 genes with decreased levels. Northern blot analysis revealed that genes for OX-40 ligand, MHC class II HLA-DR-alpha and beta-microglobulin showed predominant increase followed by tyrosine 3'-monooxygenase, proteolipid protein, tyrosine beta, and apolipoprotein D. Genes that underwent reduced expression included cytochrome c oxidase subunit I, ATP synthase isoform, cytochrome b, ATPase 9, and at least one unidentified gene. CONCLUSIONS: Our results, particularly the increased detection of OX-40 ligand and MHC class II-alpha and beta-2 microglobulin, support the finding that lymphocytes persist in latently-infected trigeminal ganglia (TGs). These results also suggest a role of the immune system in HSV-1 recurrences. In addition, we have detected genes associated with neuronal disorders, OX-40L, proteolipid protein, and apolipoprotein D.
PURPOSE: The aim of the present study was to identify the genes that express in the nervous system when herpes simplex virus type 1 (HSV-1) reactivates from latency by using subtraction cloning following epinephrine iontophoresis. METHODS: The corneas of New Zealand White rabbits (2 to 2.5 kg) were topically inoculated with 5 x 10(5) PFU of plaque-purified HSV-1 strain McKrae. Corneal infection was monitored by slit lamp examination (SLE) on days 3-7 after infection. At 30 d post-infection, eyes were assessed for infectious virus by ocular swabs. Disappearance of virus in ocular swabs signified latency for HSV-1. Latently-infected rabbits were subjected to transcorneal iontophoresis. The messenger RNA (mRNA) was isolated from the trigeminal ganglia and used for subtraction cloning and subtraction library synthesis. RESULTS: Our results indicate 18 genes with increased expression and 6 genes with decreased levels. Northern blot analysis revealed that genes for OX-40 ligand, MHC class II HLA-DR-alpha and beta-microglobulin showed predominant increase followed by tyrosine 3'-monooxygenase, proteolipid protein, tyrosine beta, and apolipoprotein D. Genes that underwent reduced expression included cytochrome c oxidase subunit I, ATP synthase isoform, cytochrome b, ATPase 9, and at least one unidentified gene. CONCLUSIONS: Our results, particularly the increased detection of OX-40 ligand and MHC class II-alpha and beta-2 microglobulin, support the finding that lymphocytes persist in latently-infected trigeminal ganglia (TGs). These results also suggest a role of the immune system in HSV-1 recurrences. In addition, we have detected genes associated with neuronal disorders, OX-40L, proteolipid protein, and apolipoprotein D.
Authors: Nicholas Farley; David I Bernstein; Fernando J Bravo; Julie Earwood; Nancy Sawtell; Rhonda D Cardin Journal: Antiviral Res Date: 2010-02-16 Impact factor: 5.970