In Projects 1-3, the investigators will analyze the elements of the innate responses induced in the presenceof flagellin or the viral vaccine vectors; develop approaches to modulate the elements and magnitude ofthese responses; and evaluate the impact of the induced innate responses on the generation of Yersinia)estis F1 and V antigen specific antibodies. Although the measurement of IgG titers against Y. pestis F1]nd V antigens is of obvious importance in assessing the efficacy of flagellin and the viral vaccine vectors,we believe that it is essential to determine the quality of the memory immune response in terms of its abilityto protect animals against challenge with Y. pestis. Therefore, we propose to establish an Animal ChallengeCore Laboratory of the Program, that will provide a uniform and consistent animal model for cross-evaluationof vaccine and adjuvant efficacy. The Core will have the following aims:
Specific Aim 1. To provideconsistent, high quality BSL3 reagents that are needed for the evaluation of adjuvants and vaccines. The Y._estis wild-type CO92, originally isolated from the sputum of a human case of pneumonic plague, will beJsed in the challenge studies. The Core will prepare frozen stocks of the bacteria and will maintained themat -80C and will prepare standardized doses of the bacteria for experiments.
Specific Aim 2. To performuniform and consistent challenge experiments using Y. pestis strain CO92 in mice. A critical aspect of thework in Projects 1-3 will be to test the efficacy of various immunization protocols using flagellin as anadjuvant or SV5 or VSV as vaccine vectors for the F1/V fusion protein. The Core will establish areproducible challenge model in BALB/c mice. The mice will be immunized with the F1/V fusion proteinaccording to the protocols established in Projects 1-3 and then will subject the mice to challenge with Y.pestis CO92 using s.c. injection or a nose-only inhalation exposure system (CH Technologies, Inc.).Measurements will include survival and bacterial burden in the lungs and spleen of animals.
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