ABSTRACT
BACKGROUND: The highly pathogenic avian influenza H5N1 virus poses a serious threat to humans. Due to its antiviral activity, antibody-based therapy is one of the possible effective countermeasures. Here, a combination of intracellular and extracellular human antibodies was investigated and showed an improved protective effect. METHODS: The scFv4F5-based intracellular antibody vectors and IgG1 extracellular antibody were constructed and expressed, respectively, and the sensitivity, specificity, and affinity of these antibodies were determined in vitro. In vivo, the protective effect of IgG1 and the combination of antibodies were tested respectively. Furthermore, the dynamics of viral replication, the related cytokines and apoptosis-related proteins were detected. RESULTS: In vitro, the expressed intracellular antibody inhibited H5N1 virus propagation and the IgG1 exhibited high specificity, sensitivity, and affinity against the H5N1 virus. In vivo, the extracellular antibody could inhibit viral propagation in a dose-dependent manner. The protective effect of IgG1 was good in a mouse model, and the survival was 100% at a dose of 15 mg/kg under infection with 100 TCID50 virus. When the intracellular antibody was pre-transfected in combination with IgG1, it had a better protective effect. The survival was 16.67% under treatment with IgG1 alone and up to 83.33% under treatment with the combination of antibodies when challenge of 500 TCID 50 virus. Furthermore, the levels of cytokines IFN-γ, IL-6, IL-10 and some apoptosis-related proteins increased. CONCLUSIONS: This antibody combination technique could be used as an appropriate and powerful alternative to antiviral therapy.
