FIG. 3.

Establishment of latency (16 days) following i.n. infection is independent of the infectious dose. B6 mice were infected i.n. with the indicated doses of γHV68, and 16 days later, splenocytes were evaluated. Cells from five mice were pooled and analyzed for each experiment (n = 2). (A) Ex vivo reactivation from latency. A limiting-dilution ex vivo reactivation assay was used to determine the frequency of peritoneal cells and splenocytes reactivating from latency, as described in the legend to Fig. 1A. (B) Preformed infectious virus. Duplicate samples from the reactivation assay were mechanically disrupted to test for the presence of preformed virus, as described in the legend to Fig. 1B. The presence of preformed virus in splenocytes from the 40-PFU sample group corresponded to the productive infection of, at most, 1 in 500,000 cells (0.2 PFU at 100,000 cells = 1 PFU in 500,000 cells). CPE, cytopathic effect.