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

Brazilian Journal of Microbiology

Ekaterina A. Durymanova Ono²  Keila Iamamoto²  Juliana G. Castilho¹  Pedro Carnieli Jr.¹  Rafael De Novaes Oliveira¹  Samira M. Achkar¹  Maria L. Carrieri¹  Ivanete Kotait¹  Paulo E. Brandão² 

[1],Universidade de São Paulo Faculdade de Medicina Veterinária São Paulo SP ,Brazil

关键词: rabies;    RNA interference;    siRNAs;    antiviral;    treatment;   

DOI  :  10.1590/S1517-83822013005000050

来源: SciELO

【 摘 要 】

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.

【 授权许可】

CC BY-NC   
 All the contents of this journal, except where otherwise noted, is licensed under a Creative Commons Attribution License