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Combined DNA - Antigen based immunization for the development of vaccines: the Hepatitis C virus model

Obiettivo



Recent advances in molecular biology and cellular and molecular immunology have triggered vaccinology to enter new phases of development. In particular, very promising data have accumulated with regards to the new vaccination mode that is the direct injection of plasmid DNA into a host for induction of anti- viral, parasitic, bacterial and tumor immunity. Concomitant to this development, the remaining lack of efficient antiviral treatment to control one of the most medically and economically important viral associated disease in Europe, due to the hepatitis C virus (ECV), renders particularly acute the development of a vaccine against this agent.
The focus of this study is to evaluate the potential offered by the combination of DNAand purified antigen-based vaccination for induction of antiviral immunity. This is a proof of principle which can be used as a vaccine approach for many diseases. The model system proposed in this program is HCV and the animal model the chimpanzee.
We propose to: immunize animals by means of direct injection of plasmid DNA (primoimmunization) followed by vaccination (booster injection) with highly purified recombinant antigens. Immunogens will be specific of the glycoproteins El and E2 of HCV with 1) DNA immunogens being di-cistronic plasmids expressing fuJl-length and truncated forms of El and E2 and 2) purified recombinant proteins (El and E2) expressed and highly purified using recombinant vaccinia viruses.
Extensive analysis of the humoral responses (antibody titers and reactivity analyzed by Western blotting analysis and immunoflorescence studies, peptide-based epitope mapping) and cellular responses (including T-helper (TH) and cytotoxic T cell (CTL) responses) will be analyzed following each DNA and protein injection. Animals will be, in a second phase of the program, challenged with an homologous HCV isohte. Such isolate will be the prototype H-strain of HCV belonging to the subtype la group of viruses. Such viral subtype is, with the subtype lb, the most prevalent viral type existing in Europe. If animals are protected, the program will be extended to evaluate challenge with a heterologous isolate.
Evaluation of the combination of these two modes of immunization (i.e. DNA-based and antigen-based) performed in a large susceptible animal model such as proposed here, has not yet been reported neither in the field of genetic imrmlnization nor with respect to highly mutable infectious agents such as HCV against which traditional vaccines offer only poor performance. The different participating groups have strong and worldwide recognized expertise in the field of protein expression and purification (Participant 2), DNA-based immunization specific for HCV (Participant 1, coordinator) and chimpanzee vaccination (Participant 3). Participants 2 and 3 are already involved in chimpanzee-based protocols for the development of a classical HCV vaccine (via the funding of a EC program), while participants 1 and 2 are participating in another EC supported project focused on the evaluation of DNA vaccination.

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Coordinatore

INSTITUT NATIONAL DE LA SANTE ET DE LA RECHERCHE MEDICALE
Contributo UE
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Indirizzo
Cours Albert Thomas 151 ADR 5 - Rhône-Alpes, Auve
69424 LYON
Francia

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Partecipanti (2)

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