Kaposi’s Sarcoma-Associated Herpesvirus Subversion of the Anti-Inflammatory Response in Human Skin Cells Reveals Correlates of Latency and Disease Pathogenesis

Figure 6

Early activation of NF-κB signaling during acute infection with rKSHV.219 is sustained in MeWo cells, but subsequently limited in Mel1700 cells following long-term infection. (a) MeWo and Mel1700 cells either mock-infected, treated for 20 min. with TNF-α, or infected with rKSHV.219 for 20 and 60 min. were used in western blot analysis for phosphorylated NF-κB p65. Mock-infected cell lysates were used to determine baseline levels of phosphorylated NF-κB p65, whereas lysates from cells treated with TNF-α served as positive controls for activation of NF-κB p65. Total NF-κB protein was used as an internal loading control. (b) Total RNA from half of the same cells used in (a) was used for RT-PCR amplification of the NF-κB-controlled ICAM-1 gene. (c) MeWo cells (top) and Mel1700 cells (bottom) either mock-infected (control), TNF-α-treated, or infected with rKSHV.219 for 0.3 h, 1 h, 3 h, or 6 h were used in western blot analysis for phosphorylated NF-κB p65. (d) Western blot analysis of the phosphorylation levels of key components of the NF-κB-signaling pathway in long-term cultures of uninfected (−) and infected (+) MeWo and Mel1700 cells.