Adenovirus-mediated RNA interference against foot-and-mouth disease virus infection both in vitro and in vivo.
Foot-and-mouth disease virus (FMDV) infection is responsible for the heavy economic losses in stockbreeding each year. Because of the limited effectiveness of existing vaccines and antiviral drugs, the development of new strategies is needed. RNA interference (RNAi) is an effective means of suppressing virus replication in vitro. Here we demonstrate that treatment with recombinant, replication-defective human adenovirus type 5 (Ad5) expressing short-hairpin RNAs (shRNAs) directed against either structural protein 1D (Ad5-NT21) or polymerase 3D (Ad5-POL) of FMDV totally protects swine IBRS-2 cells from homologous FMDV infection, whereas only Ad5-POL inhibits heterologous FMDV replication. Moreover, delivery of these shRNAs significantly reduces the susceptibility of guinea pigs and swine to FMDV infection. Three of five guinea pigs inoculated with 10(6) PFU of Ad5-POL and challenged 24 h later with 50 50% infectious doses (ID50) of homologous virus were protected from the major clinical manifestation of disease: the appearance of vesicles on the feet. Two of three swine inoculated with an Ad5-NT21-Ad5-POL mixture containing 2 x 10(9) PFU each and challenged 24 h later with 100 ID50 of homologous virus were protected from the major clinical disease, but treatment with a higher dose of adenovirus mixture cannot promote protection of animals. The inhibition was rapid and specific because treatment with a control adenovirus construct (Ad5-LacZ) expressing Escherichia coli galactosidase-specific shRNA showed no marked antiviral activity. Our data highlight the in vivo potential of RNAi technology in the case of FMD.
State Key Laboratory of Genetic Engineering, Institute of Genetics, School of Life Science, Fudan University, Shanghai 200433, People's Republic of China.
Weizao
W
Chen
Mingqiu
M
Liu
Ye
Y
Jiao
Weiyao
W
Yan
Xuefeng
X
Wei
Jiulian
J
Chen
Liang
L
Fei
Yang
Y
Liu
Xiaoping
X
Zuo
Fugui
F
Yang
Yonggan
Y
Lu
Zhaoxin
Z
Zheng
D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D002460:Cell Line; D006224:Cricetinae; D003673:Defective Viruses / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005536:Foot-and-Mouth Disease / Q000517:prevention & control*; D029544:Foot-and-Mouth Disease Virus / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification / Q000502:physiology; D005822:Genetic Vectors; D016679:Genome, Viral; D006168:Guinea Pigs; D011558:Herpesvirus 1, Suid / Q000302:isolation & purification; D006801:Humans; D007271:Injections, Intradermal; D007273:Injections, Intramuscular; D007700:Kinetics; D008297:Male; D009500:Neutralization Tests; D016366:Open Reading Frames; D034622:RNA Interference; D012367:RNA, Viral / Q000008:administration & dosage*; D011995:Recombination, Genetic; D012703:Serotyping; D013552:Swine; D014779:Virus Replication
D012367:RNA, Viral