Inhibitory effects of native and recombinant full-length camel lactoferrin and its N and C lobes on hepatitis C virus infection of Huh7.5 cells

Abstract

Lactoferrin has been suggested to have antiviral activity against hepatitis C virus (HCV). The objective of this study was to compare the effects of recombinant camel lactoferrin (rcLf), native camel lactoferrin (ncLf) and their N and C fragments on HCV infection in Huh7.5 cells. ncLf was purified from camel milk and N and C lobes were generated proteolytically. rcLf and its fragments were synthesized using a Bac-to-Bac baculovirus expression system. All proteins except the C lobe of rcLf were soluble. The inhibitory effects on HCV entry into Huh7.5 cells were evaluated by incubation of HCV with Lf prior to infection or pre-treatment of the cells with Lf prior to infection. The inhibitory effect on HCV amplification in Huh7.5 cells was determined by Lf treatment of HCV-infected cells. Nested RT-PCR was performed to amplify intracellular HCV 59 non-coding RNA sequences. rcLf and ncLf and their fragments could prevent HCV entry into Huh7.5 cells by direct interaction with the virus and inhibited virus amplification in Huh7.5 cells. Therefore, the N and C lobes of ncLf are sufficient to elicit anti-HCV effects in Huh7.5 cells. rcLf and its N lobe displayed similar HCV inhibitory effects to their native counterparts and may constitute an efficient and cost-effective approach for potential clinical applications.