Examples of quantitation during HDV genome replicationa

Experimental systemNo. of δAg per average cell (105)No. of δRNA per average cell (105)δAg/δRNA ratio
Woodchuck liver, infected
    Animal no. 1730.66112
    Animal no. 2610.6494
HuH7 cells, transiently transfected with RNA
    Day 3190.07266
    Day 5950.27359
    Day 92400.59410
    Day 15850.29292
    Day 209.30.04210
HuH7 cell clones stably transfected with DNA
    Clone 88.70.052167
    Clone 12180.060306
    Clone 145.20.026200
Mouse liver, transfected
    Day 52.30.020118
    Day 96.80.034200
    Day 150.390.02416
    Day 201.40.02848
    Day 300.110.00338
Mouse skeletal muscle, transfected
    Day 8<0.720.018<40
    Day 200.720.03918
    Day 400.960.03726
    Day 80<0.720.004<180
  • a Five different methods for the initiation of HDV genome replication are shown here. Woodchucks already infected with woodchuck hepatitis virus were superinfected with serum from an HDV-infected animal, as previously described (29). Transient transfection of HuH7 cells with antigenomic HDV RNAs was as previously described (27). The three HuH7 cell clones obtained by stable transfection of HuH7 cells with pSVL(D3) (20) have been described previously (9). Mice were transfected with the HDV cDNA construct pDL456 (22) by a hydrodynamics-based procedure to achieve replication in the liver (6). After moving the HDV insert of pDL456 from pSVL (Pharmacia) to pcDNA3 (Invitrogen), the resulting construct, pJC126, was directly injected into mouse skeletal muscle by following a previously described procedure (30). Indicated for each sample are the deduced numbers of molecules of δAg and genomic δRNA per average cell along with the molar ratio. Quantitation of δAg refers to both small and large forms.