Abstract
Abstract: :
Purpose:To determine changes in gene expression in HSV-1 latently infected mouse trigeminal ganglia (TGs) following treatment with cyclophosphamide and dexamethasone. Methods: Scarified corneas of BALB/c mice were inoculated with 2.5 X 104 PFU HSV-1 strain McKrae. Four weeks after inoculation, cyclophosphamide and dexamethasone were intravenously injected to induce HSV-1 reactivation. Uninfected mice were also treated with the drugs. Four groups of mice were studied: 1) uninfected, not treated, 2) uninfected, drug treated, 3) latently infected, not treated, 4) latently infected, drug treated. Poly A+ mRNA from the TGs of each group was reverse transcribed, labeled with 32P, incubated on a 1185-gene array membrane, and analyzed by phosphorimaging. To confirm microarray results, semi-quantitative RT-PCR for selected genes was performed. Results: A statistically significant increase in the expression of 10 genes (prostaglandin E2 receptor EP4 subtype, insulin promoter factor 1, glutathione S-transferase mu 2, cyclin D2, peripherin, plasma glutathione peroxidase, methyl CpG-binding protein 2, retinal S-antigen, ErbB2 proto-oncogene, guanine nucleotide-binding protein alpha stimulating activity polypeptide) and decreased transcriptional activity of 8 genes (peripheral myelin protein 22, decorin, transcription factor AP-1, dystroglycan 1, myelin protein 0, mitogen-activated protein kinase 3, prothymosin beta 4, brain lipid-binding protein) were documented in the ganglia of latently infected immunosuppressed mice. The results obtained by semi-quantitative RT-PCR were similar to those obtained by microarray analysis. Conclusion: Changes in gene expression in the prostaglandin pathway, transcription factors, and enzymes in the cell cycle may be important in HSV-1 reactivation induced by immunosuppression.
Keywords: 425 herpes simplex virus • 417 gene/expression