FIGURE 3.

Defining the steps of the HBV life cycle targeted by IL-1β. A, HepaRG cells were pretreated with IL-1β or heparin for 3 h and then infected with HBV in the presence (A, panel a) or absence (A, panel b) of IL-1β or heparin for 16 h. HBV infection was monitored with HBs protein secretion from the infected cells. Only pretreatment with IL-1β and not heparin could inhibit HBV infectivity. d, day. B, HepaRG cells were pretreated with IL-1β or left untreated (−) for the indicated time (h) and infected with HBV without IL-1β. Anti-HBV activity was amplified by a prolonged treatment time. C, panel a, HepaRG cells were pretreated with 10 ng/ml IL-1β, 100 IU/ml IFNα, or 1 μm lamivudine for 3 h, followed by infection with HBV for 16 h in the absence of cytokines (pretreatment). C, panel b, HepaRG cells were infected with HBV for 16 h without pretreatment. After washing out the input virus, cells were cultured in normal medium for the first 8 days and then cultured with IL-1β, IFNα, or lamivudine for the following 4 days (post-treatment). HBV DNA in the cells was measured by real time PCR. IL-1β showed an anti-HBV activity in both pretreatment and post-treatment, although an anti-HBV effect of IFNα was seen only with post-treatment. D, HepAD38 cells were treated with 100 ng/ml IL-1β or 1 μm lamivudine, or left untreated for 6 days in the absence of tetracycline. HBV replication was evaluated by measurement of HBV DNA in the medium. E, HepaRG cells were pretreated with IL-1β, lamivudine, or heparin for 3 h or left untreated and infected with HBV for 16 h in the presence or absence of each compound. After trypsinization and extensive washing of the cells, cellular DNA was immediately recovered to detect HBV DNA. HBV DNA at 16 h post-infection was decreased by treatment with IL-1β but not lamivudine.