Generating antibodies against the native form of the human prion protein (hPrP) in wild-type animals: A comparison between DNA and protein immunizations

Generation of therapeutic antibodies against human proteins is hampered by the difficulty of obtaining large quantities of correctly folded immunogens when following classic immunization procedures. Here we compared several genetic immunization protocols for their potential ability to generate high levels of antibodies against proteins expressed in their native form. We chose as a model the prion protein (PrP) because it has been demonstrated that the recognition of the native conformation of PrP is an absolute prerequisite for anti-PrP antibodies to be used as therapeutic tools for prion diseases, a group of lethal neurodegenerative disorders. We designed two human PrP-DNA vectors, containing or not a stimulatory T cell epitope, which were injected into mice following four different protocols: in the naked form with or without electroporation, or protected by cationic polymers or block copolymers. For comparison, other animals received conventional injections of recombinant human PrP with Freund's adjuvant or alum. We found that genetic immunization, carried out especially through DNA electroporation and, to a lesser extent, through injection of block copolymer-protected DNA, was able to generate high amounts of antibodies recognizing native PrP as expressed on the cell surface. Conversely, protein immunizations led to very high levels of antibodies against PrP immobilized on microtiter plates, but unable to recognize the native cell membrane-bound PrP. This clearly demonstrates the usefulness of genetic immunization, when performed under well defined conditions, in raising antibodies to native proteins. These results are of interest not only in view of passive immunotherapy of prion diseases, but also, more generally, in view of generating antibodies to human membrane proteins for immunotherapeutic or immunodiagnostic purposes. (C) 2008 Elsevier B.V. All rights reserved.