Abstract
Abstract: :
Purpose: To identify genes whose expression in trigeminal ganglia is differentially regulated early after ocular inoculation of HSV-1. Methods: Groups of 129S6 mice were bilaterally inoculated with a total of 1x106 PFU HSV-1 strain F or sham inoculated with PBS by corneal scarification. On day 4 PI the mice were sacrificed and right and left trigeminal ganglia were pooled from 5 mice in each group. Total ganglionic RNA was purified from the sham and HSV inoculated groups of mice and used to prepare 32P-labeled cDNA probes that were hybridized to cDNA microarrays. Semi-quantitative RT-PCR assays were used to confirm differential expression of genes, immunostaining was used to localize gene expression to specific cell types in vivo antibody mediated ablation of protein was used to assess relevance to HSV infection. Results: Analysis of Clontech cDNA microarrays revealed that murine monokine induced by interferon gamma (muMIG) was upregulated more than 100 fold at day 4 PI. muMIG was not expressed in ganglia of sham inoculated 129S6 mice or in ganglia from HSV inoculated 129S6 mice deficient in IFN-g expression, confirming the importance of IFN-g for induction of muMIG expression. Using a rabbit polyclonal antibody, expression of muMIG appeared to be confined to ganglionic neurons. To determine a role for muMIG, groups of HSV inoculated mice are being treated with either normal rabbit serum or a polyclonal muMIG antiserum and the course of infection monitored. Conclusion: Since muMIG functions to attract activated T cells, our results demonstrating early induction of this chemokine in ganglionic neurons suggest it has a potentially important role in HSV infection in vivo.
Keywords: 425 herpes simplex virus • 380 cytokines/chemokines • 417 gene/expression