Figure 7. Cytokine production in splenocytes derived from mice inoculated with a cocktail of proteins BtuB, Hia, and FlgK, B. abortus S19 lysate, ISCOMTRIX adjuvant, or PBS.

Groups of 3 C57BL/6 mice were inoculated subcutaneously twice three weeks apart and humanely euthanized 3 weeks after the last inoculation. Spleens were harvested and processed into a single cell suspension. Splenocytes were plated and stimulated with individual Brucella outer membrane proteins, S19 lysate, ISCOMATRIX adjuvant or RPMI media. Supernatants were collected 24 hrs post-stimulation and analyzed for cytokines using the Th1/Th2 bioplex kit or indirect ELISA (IL-17A). Data represent mean +/− standard error of the mean from each group of 3 mice. Statistical significance between PBS control and treatment groups was determined with one-tailed Student’s t-test (*P<0.05, **P<0.01, ***P<0.001).