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S. M. Swanson, A. S. Bertke, K. Apakupakul, A. Ma, T. P. Margolis; Differential Herpes Simplex Virus (HSV) Gene Expression in Ganglionic Neurons. Invest. Ophthalmol. Vis. Sci. 2010;51(13):3870.
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We previously demonstrated that different subtypes of neurons in the trigeminal ganglion are differentially permissive for productive infection with HSV-1 in vivo, with A5+ neurons being the least permissive. The aim of the current work was to determine whether this was also true for neurons infected in vitro.
Cultures of dissociated adult murine trigeminal ganglia were infected with HSV-1 constructs that expressed enhanced green fluorescent protein (EGFP) from immediate-early (ICP0), early (gB) and late (gC) gene promoters at an multiplicity of infection of 30. Infected cultures were co-labeled with MAb A5 and evaluated by fluorescence microscopy.
Neuronal cultures contained proportions of A5+ neurons similar to that seen in vivo. Expression of HSV-1 ICP0, gB and gC was first noted at 2, 3 and 4 hours post-infection (p.i.), respectively, with gene expression reaching a plateau at 6, 8 and 10 hours p.i., respectively. At 6 hours p.i. 9% of the cultured neurons were A5+, but only 1% of the ICP0 expressing neurons were A5+. At 8 hours p.i. 8% of the cultured neurons were A5+, but only 2% of the gB expressing neurons were A5+. At 10 hours p.i. 10% of the cultured neurons were A5+, but only 2% of gC expressing neurons were A5+. In contrast nearly all microglia expressed all three classes of genes.
Microglia were much more permissive for productive viral infection than neurons. As compared to the overall population of cultured ganglionic neurons, A5+ neurons are less permissive for productive infection with HSV-1. Differential permissiveness for productive infection with HSV-1 appears to be regulated at, or before, expression of the viral immediate early genes.
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