TABLE 3.

Examples of quantitation during HDV genome replication^a

Experimental system	No. of δAg per average cell (105)	No. of δRNA per average cell (105)	δAg/δRNA ratio
Woodchuck liver, infected
Animal no. 1	73	0.66	112
Animal no. 2	61	0.64	94
HuH7 cells, transiently transfected with RNA
Day 3	19	0.07	266
Day 5	95	0.27	359
Day 9	240	0.59	410
Day 15	85	0.29	292
Day 20	9.3	0.04	210
HuH7 cell clones stably transfected with DNA
Clone 8	8.7	0.052	167
Clone 12	18	0.060	306
Clone 14	5.2	0.026	200
Mouse liver, transfected
Day 5	2.3	0.020	118
Day 9	6.8	0.034	200
Day 15	0.39	0.024	16
Day 20	1.4	0.028	48
Day 30	0.11	0.003	38
Mouse skeletal muscle, transfected
Day 8	<0.72	0.018	<40
Day 20	0.72	0.039	18
Day 40	0.96	0.037	26
Day 80	<0.72	0.004	<180

^aFive 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.