A Single Nasal Dose Vaccination with a Brucella abortus Mutant Potently Protects against Pulmonary Infection

The Brucella abortus double-mutant (ΔznuA ΔnorD Brucella abortus-lacZ [znBAZ]) was assessed for its protective efficacy after vaccination with a single nasal dose. Superior protection was achieved in znBAZ-vaccinated mice against pulmonary, wild-type B. abortus 2308 challenge when compared with conventional livestock Brucella abortus vaccines, the smooth S19 (smooth B. abortus strain 19 vaccine) and rough RB51 (rough mutant vaccine strain of B. abortus) strains. Nasal znBAZ vaccination reduced splenic and lung colonization by wild-type brucellae by >3-4 logs. In contrast, S19 reduced lung colonization by only 32-fold, and RB51 failed to reduce colonization. One profound attribute of znBAZ vaccination was the >3-fold increase in pulmonary CD8+ T cells when compared with other vaccinated groups. S19 vaccination increased only CD4+ T cells. All vaccines induced IFN-γ and TNF-α production by CD4+ T cells, but only znBAZ vaccination enhanced the recruitment of polyfunctional CD8+ T cells, by >100-fold. IL-17 by both CD4+ and CD8+ T cells was also induced by subsequent znBAZ vaccination. These results demonstrate that, in addition to achieving protective immunity by CD4+ T cells, CD8+ T cells, specifically resident memory T cells, also confer protection against brucellosis. The protection obtained by znBAZ vaccination was attributed to IFN-γ-producing CD8+ T cells, because depletion of CD8+ T cells throughout vaccination and challenge phases abrogated protection. The stimulation of only CD4+ T cells by RB51- and S19-vaccinated mice proved insufficient in protecting against pulmonary B. abortus 2308 challenge. Thus, nasal znBAZ vaccination offers an alternative means to elicit protection against brucellosis.