Fig 6.

Efficient reactivation of HCMV from MoLCs is maturation dependent and enhanced by IL-6. (A) CD14⁺ monocytes TB40/e (lanes 1 and 2) or mock (lanes 3 and 4) infected were analyzed for RNA expression at 3 days postinfection. RNA with (+) or without (−) prior RT was amplified in UL138, IE72, and actin-specific PCRs. For the IE72 PCR, an HCMV DNA PCR-positive control was included to confirm that the PCR had worked (lane 5). (B) RNA isolated from immature MoLCs either mock treated (lane 1) or treated with IL-6 (lane 2), IL-8 (lane 3), LPS (lane 4), LPS plus neutralizing IL-6 antibody (lane 5), or LPS plus neutralizing IL-8 antibody (lane 6) was (+) or was not (−) subjected to RT and then amplified in an IE72 or actin PCR. (C and D) MoLCs were cocultured with fibroblasts to assay HCMV reactivation. After 10 days, the cultures were analyzed for evidence of plaque formation (C) and HCMV reactivation by inoculating fresh monolayers of fibroblasts with 50 μl of the supernatant and staining for IE gene expression 24 h postinfection as an indicator of infectious virus in the supernatant (D).