Combination of specific single chain antibody variable fragment and siRNA has a synergistic inhibitory effect on the pro
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RESEARCH
Open Access
Combination of specific single chain antibody variable fragment and siRNA has a synergistic inhibitory effect on the propagation of avian influenza virus H5N1 in chicken cells Shuang Wang1, Peng Zhang2, Fei He1, Ji-Gui Wang1, Jia-Zeng Sun1, Zhi-Li Li1, Bao Yi1, Ji Xi1, Ya-Ping Mao1, Qiang Hou1, Dao-Li Yuan1, Zi-Ding Zhang1 and Wei-Quan Liu1*
Abstract Background: The avian influenza virus (AIV) causes frequent disease with high morbidity and mortality. RNA interference (RNAi) has been shown to provide an effective antiviral defense in animals, and several studies have focused on harnessing small interfering RNAs (siRNAs) to inhibit viral infections. In addition, single chain variable fragments (scFvs) contain the complete antigen binding site, and specific scFvs can bind to and neutralize viruses. Results: Fourteen positive scFvs were selected by the yeast two-hybrid system. Using molecular docking technology, we selected the three highest affinity scFvs for further functional validation. Results of indirect ELISA and IFA showed that all three scFvs could bind to FJ13 strain and had neutralizing activity, decreasing the viral infectivity markedly. Chicken fibroblastic DF-1 cells were transfected with scFvs in combination with siRNA-NP604 (an siRNA of anti-AIV NP protein previously reported). Following infection with FJ13 virus, copy numbers of the virus were significantly reduced from 12 h to at least 60 h post-infection compared to that achieved in cells transfected with scFv or siRNA-NP604 separately. Conclusions: A novel combination of antiviral siRNAs expressed in chicken cells and chicken antibody single-chain variable fragments (scFvs) secreted from the cells has a synergistic inhibitory effect on the avian influenza viral proliferation in vitro. Intracellular application of scFvs and anti-viral siRNA may provide a new approach to influenza prevention and treatment. Keywords: AIV H5N1, scFv, siRNA, Synergistic inhibitory effect
Introduction Avian influenza viruses (AIVs) are important pathogens affecting the poultry industry worldwide, with some infecting humans with a high fatality rate [1]. Due to the diversity and propensity for inter-species transmission, AIVs present a challenge for vaccine developers [2,3]. RNA interference (RNAi) has been shown to provide an effective antiviral defense in animals, and several studies have focused on harnessing small interfering RNAs (siRNAs) to inhibit viral infections [4-8]. An siRNA-based * Correspondence: [email protected] 1 State Key Laboratory of Agrobiotechnology, Department of Biochemistry and Molecular Biology, College of Biological Sciences, China Agricultural University, Beijing 100193, China Full list of author information is available at the end of the article
microbicide has been shown to protect mice from lethal herpes simplex virus 2 infection or transmission [9], and siRNAs specific for conserved regions of influenza virus genes prevented and even treated influenza viral infection in mice [6]. In 2011, Lyall et al. [3] reporte
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