A DNA Vaccine Strategy for Effective Antibody Induction to Pathogen-Derived Antigens
DNA-based vaccines are currently being developed for treating a diversity of human diseases including cancers, autoimmune conditions, allergies, and microbial infections. In this chapter, we present a general protocol that can be used as a starting point
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1. Introduction DNA vaccines have been the subject of intense investigation over the last 2 decades and the technology provides an adaptable and powerful means for activating both innate and adaptive immune pathways and allowing multiple vaccines to be made and tested quickly and cost-effectively (1). The concept is very simple: DNA encoding the antigen of choice is inserted into a bacterial plasmid, with gene expression usually driven by a strong viral promoter. Delivery of the plasmid vaccine into muscle or skin cells leads to antigen production and presentation to the immune system, and both antibody and cell-mediated immune responses can be effectively induced. In addition, the plasmid also has excellent intrinsic adjuvant properties: the dsDNA acts as a pathogen-associated molecular pattern that can trigger a range of cellular receptors,
Myron Christodoulides (ed.), Neisseria meningitidis: Advanced Methods and Protocols, Methods in Molecular Biology, vol. 799, DOI 10.1007/978-1-61779-346-2_22, © Springer Science+Business Media, LLC 2012
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leading to type I interferon production and an inflammatory response by cells of the innate immune system. Thus, plasmid DNA not only delivers antigen, but engages multiple routes to activate innate immunity. DNA-based vaccines are in development for infectious diseases and as therapies against autoimmune diseases, allergy and cancer, and several DNA vaccines are already licensed for veterinary use (2). Importantly, DNA vaccines can induce antibody responses against bacterial pathogens where humoral immunity is believed to be essential, e.g. against peptide mimotopes of meningococcal group B and group C polysaccharides (3, 4) and porin polypeptide (5) and Streptococcus pneumoniae capsular polysaccharide (6) and surface antigens (7), Borrelia burgdorferi outer surface proteins (8, 9), Brucella outer membranes (OM) (10) and OM porin OprF of Pseudomonas aeruginosa (11). In this chapter we present a general protocol that can be used as a starting point for developing DNA vaccines to pathogen-derived antigens, using Neisseria meningitidis as an example. In addition, for antigens that can be considered as poorly immunogenic, such as short pathogen-derived polypeptides (5), we also provide a protocol for adopting a fusion gene-based vaccine design similar to the haptencarrier system to increase the potency of our DNA vaccines (2). This strategy links highly immunostimulatory sequences, such as the fragment C (FrC) sequence of tetanus toxin, to weak target antigens within the DNA vaccine format. This fusion gene vaccine strategy can induce high-affinity antibodies to the linked target antigen through several mechanisms, the most important of which is the cognate CD4+ T-cell help provided by linked immunostimulatory elements. Finally, we provide a protocol for efficient delivery of DNA vaccine, based on intramuscular injection followed by electroporation, which delivers an immediate electrical current across the injection site. This causes an in
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