In this study, we examine the potential of a combinatorial vaccine consisting of the lead-candidate antigens for the next generations of vaccines against anthrax (rPA) and plague (F1-V) with the specific objective of determining synergy or interference between the vaccine components when they are administered separately or together by both traditional parenteral immunization (SC) and mucosal immunization (IN) in the presence of appropriate adjuvants. The most significant findings of the study reported here are that (1) a combinatorial vaccine consisting of equal amounts of F1-V and rPA administered SC is effective at eliciting a robust serum and bronchoalveolar lavage (BAL) antigen-specific IgG and IgG1 response against both antigens in immunized animals, and when administered IN, a robust antigen-specific IgG2a response in the serum and BAL is also induced; (2) there were few instances where either synergy or interference was observed in the combined vaccine administered by either route and those differences occurred soon after the final immunization and were not sustained over time; (3) IN immunization was as effective as SC immunization for induction of antigen-specific serum and BAL antibody responses using the same amount of antigen; (4) the IgG1/IgG2a ratios suggest a strongly biased Type 2 response following SC immunization, while IN immunization produced a more balanced Type 1/Type 2 response; (5) the IgG1/IgG2a ratio was influenced by the route of immunization, the adjuvant employed, and the nature of the antigen. As with previously published studies, there were still detectable levels of circulating anti-F1-V and anti-rPA even 6 months post-primary immunization. These studies provide important insights into the development of new generation biodefense vaccines.
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