Abstract
Interferon alpha (IFNα) is the first line treatment for chronic hepatitis B and C. In order to test new IFNα delivery systems and investigate the function of this cytokine in the woodchuck model, the best animal model of chronic hepatitis B, we produced and purified recombinant woodchuck IFNα and used it to produce monoclonal antibodies. wIFNα5 was cloned in a prokaryotic expression system, expressed as His-tagged protein and then purified. The rwIFNα5 protein was found to induce STAT-3 phosphorylation, to enhance 2′,5′-oligoadenylate synthetase mRNA levels and to possess a potent antiviral activity. Two monoclonal antibodies were obtained through immunization of rats with rwIFNα5. Both recognized rwIFNα5 in western blot analysis and one was able to neutralize the antiviral activity of the rwIFNα5 and lymphoblastoid IFNα preparations. Finally, a capture rwIFNα5 ELISA was developed using both antibodies. In summary, the tools generated in this study will allow different forms of IFNα delivery as well as different combination therapies in woodchuck hepatitis virus infection to be tested, thus providing useful information for the design of new strategies to treat chronic hepatitis B in humans.
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