FIG 5.

Reactivation and latent viral load are increased in the absence of endosomal TLR. Mice of the indicated genotypes were infected with 2 × 10⁶ PFU of MHV68 intravenously. At 21 days postinfection, spleens were harvested, and single-cell splenocyte suspensions pooled from multiple mice were prepared. (A to C) Three-fold serial dilutions of splenocytes were cocultured with NIH 3T3 fibroblasts for 2 weeks, and reactivation was determined based on the presence of cytopathic effect in the NIH 3T3 monolayer. Curve fit lines were derived from nonlinear regression analysis based on the variable slope sigmoidal dose-response equation. The regression line intersect was set to 63.2% based on the Poisson distribution. (D) DNA was isolated from pooled splenocytes, and the gB copy number was determined relative to that of β2M and scaled to the average of the B6N or B6J WT control as appropriate. For each set of pooled splenocytes, copy numbers were determined for 3 to 4 replicates. Data points represent individual replicates. Log-transformed data (data points for >10³ splenocytes in panels A to C and all samples in panel D) were analyzed by two-tailed unpaired t test (*, P < 0.05; **, P < 0.01; ***, P < 0.001).