title: 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.
authors: Faculdade de Medicina Veterinária, Universidade de São Paulo, São Paulo, SP, Brazil. ; Instituto Pasteur, São Paulo, SP, Brazil.
authors: Ekaterina A
authors: EA
authors: Durymanova Ono
authors: Faculdade de Medicina Veterinária, Universidade de São Paulo, São Paulo, SP, Brazil. ; Instituto Pasteur, São Paulo, SP, Brazil.
authors: Keila
authors: K
authors: Iamamoto
authors: Instituto Pasteur, São Paulo, SP, Brazil.
authors: Juliana G
authors: JG
authors: Castilho
authors: Instituto Pasteur, São Paulo, SP, Brazil.
authors: Pedro
authors: P
authors: Carnieli
authors: Instituto Pasteur, São Paulo, SP, Brazil.
authors: Rafael
authors: R
authors: de Novaes Oliveira
authors: Instituto Pasteur, São Paulo, SP, Brazil.
authors: Samira M
authors: SM
authors: Achkar
authors: Instituto Pasteur, São Paulo, SP, Brazil.
authors: Maria L
authors: ML
authors: Carrieri
authors: Instituto Pasteur, São Paulo, SP, Brazil.
authors: Ivanete
authors: I
authors: Kotait
authors: Faculdade de Medicina Veterinária, Universidade de São Paulo, São Paulo, SP, Brazil.
authors: Paulo E
authors: PE
authors: Brandão
mesh_terms: D000818:Animals; D000998:Antiviral Agents / Q000378:metabolism*; D002460:Cell Line; D006224:Cricetinae; D004195:Disease Models, Animal; D051379:Mice; D019590:Nucleocapsid Proteins / Q000037:antagonists & inhibitors; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism*; D011818:Rabies / Q000188:drug therapy*; D011820:Rabies virus / Q000187:drug effects* / Q000502:physiology*; D016019:Survival Analysis; D019562:Viral Load; D014776:Virus Cultivation; D014779:Virus Replication / Q000187:drug effects*
chemical_list: D000998:Antiviral Agents; D019590:Nucleocapsid Proteins; D034741:RNA, Small Interfering; C099601:nucleocapsid protein, Rabies virus
keywords: RNA interference; antiviral; rabies; siRNAs; treatment