In vitro and in vivo inhibition of rabies virus replication by RNA interference

Abstract

Rabies is a zoonotic disease that affects all mammals and leads to more than 55,000 human deaths every year, caused by rabies virus (RABV) (Mononegavirales: Rhabdoviridae: Lyssavirus). Currently, human rabies treatment is based on the Milwaukee Protocol which consists on the induction of coma and massive antiviral therapy. The aim of this study was to assess the decrease in the titer of rabies virus both in vitro and in vivo using short-interfering RNAs. To this end, three siRNAs were used with antisense strands complementary to rabies virus nucleoprotein (N) mRNA. BHK-21 cells monolayers were infected with 1000 to 0.1 TCID50 of PV and after 2 hours the cells were transfected with each of tree RNAs in separate using Lipofectamine-2000. All three siRNAs reduced the titer of PV strain in a least 0.72 logTCID50/mL and no cytotoxic effect was observed in the monolayers treated with Lipofectamine-2000. Swiss albino mice infected with 10.000 to 1 LD of PV strain by the intracerebral route were also transfected after two hours of infection with a pool 3 siRNAs with Lipofectamine-2000 by the intracerebral route, resulting in a survival rate of 30% in mice inoculated with 100 LD50, while the same dose led to 100% mortality in untreated animals. Lipofectamine-2000 showed no toxic effect in control mice. These results suggest that intracerebral administration of siRNAs might be an effective antiviral strategy for rabies.

Short interfering RNAs targeting a vampire-bat related rabies virus phosphoprotein mRNA

“…In a previous report by some of the authors of the present article, 8 the use of N mRNA targeted siRNAs was shown to reduce the titers of the PV RABV strain in at least 0.72 log and a 30% survival rate was obtained in mice inoculated with PV and then administered a pool of these siRNAs. Nonetheless, as the PV is a fixed RABV strain and might not reflect the infection dynamics of a natural RABV infection, the use of a non-fixed strain as RABV-4005 can bring RNAi closer to a clinical situation of rabies.…”

“…This study was designed to assay the effects of short-interfering RNAs (siRNAs) against the mRNA of the P protein of a RABV strain related to Desmodus rotundus in vitro and in vivo in a post-exposure protocol as an expansion of a previous report. 8 …”

Disodium Cromolyn and Anti-podoplanin Antibodies Strongly Inhibit Growth of BHK 21/C13-derived Fibrosarcoma in a Chick Embryo Chorioallantoic Membrane Model

“…Regarding BHK-21/C13 cell response to different therapeutic agents, previous research was mainly focused on the inhibitory effects of potential or certified antiviral agents (20,21). Other drugs with different properties such as antiallergic, antitumor or antiangiogenic actions, have not been tested yet on BHK-21/C13 cell-derived sarcomas as far as we are aware.…”