Figure 10. TLR-9 dependency of type-I IFN production triggered by rodent parvoviruses in infected hPBMCs.

(A, B and C) hPBMCs were distributed into 6-well plates at 1×10⁷ cells/5 ml culture medium/well. They were immediately pre-treated, or not, for 3 hrs with the TLR-9 inhibitor ODN TTAGGG (iODN) at 2 µM and then infected or not with MVMp or H-1PV (20 PFUs/cell). In addition, some hPBMC suspensions were directly stimulated with the TLR-9 agonist ODN 2395 at 1 µM and were used as positive control experiments. Culture supernatants (A, B) as well as total DNA (C) were isolated 24 hrs later to perform ELISA for type-I IFNs and Southern blot experiments, respectively, as described previously in Figure 1 and 2. (A, B) Results are expressed as means+standard deviations of three independent experiments. Inhibitory effects of ODN TTAGGG on the stimulatory ODN 2395 were not assessed (ND). (C) (dRF, dimer replicative form; mRF, monomeric replicative form; ssDNA, single-stranded DNA genome). The blot shown is representative of 3 experiments which gave similar results.