FIG. 2.

Characterization of IL-6 induction by HSV-kinetics and effects of cycloheximide, anti-TNF-α, and UV-irradiation of the virus. RAW 264.7 cells were seeded and left overnight to settle. (A) The cells were treated with 3 × 10⁶ PFU of HSV-1 (KOS) per ml, 3 × 10⁶ PFU of HSV-2 (MS) per ml, and 10 IU of IFN-γ per ml. After 24 h, supernatants were harvested and analyzed for IL-6 by ELISA. The results are shown as means ± standard errors of the means. (B) The cells were treated as indicated with the following concentrations: 3 × 10⁶ PFU of HSV-1 (KOS) per ml or equivalent amount of UV-irradiated virus, 10 IU of IFN-γ per ml, 1,000 U of neutralizing anti-TNF-α per ml, 10 μg of CHX per ml. After 4 h of treatment total RNA was isolated. (C) The cells were treated with 3 × 10⁶ PFU of HSV-1 (KOS) per ml and 10 IU of IFN-γ per ml. After the indicated time points total RNA was isolated. (B and C) The RNA was subjected to RT-PCR using oligo(dT)₁₅ RT-priming and PCR primers specific for murine β-actin and IL-6.