pneumoniae is the use of LAB as carriers of different pneumococcal antigens. In previous studies we have demonstrated that immunization with PppA, expressed learn more as a wall-anchored protein on the surface of L. lactis, was able to induce cross-protective immunity against different pneumococcal serotypes, afforded protection against both systemic and respiratory pneumoccocal challenges, and induced

protective immunity in adult and infant mice [16]. Additionally, on the basis of previous studies, we have demonstrated that the nasal route is the best alternative for protection against a pneumococcal infection using L. lactis as adjuvant [14,15] and as antigen delivery vehicle [16,31]. This agrees with the findings of other researchers HDAC inhibitor who demonstrated the convenience of the nasal route for the immunization of mucosae against respiratory pathogens [32,33]. In this work we have assessed new immunization strategies using an inactivated recombinant bacterium by itself and in association with a probiotic strain. Analysis of the immunostimulatory properties of non-viable LAB strains showed that they depend upon the strain used, although

there is evidence indicating that viable bacteria are more effective for mucosal immunostimulation. In most cases, heat-killed strains were assessed in which differences in immunostimulation might be associated with heat-induced alteration of epitopes [34]. In order to conserve the structure of the PppA expressed in the surface of L. lactis, death was carried out by chemical inactivation. The inactivated strain proved to be effective for the induction of high levels of specific IgA and IgG antibodies in BAL and of IgG in the serum of the vaccinated young mice, which

were higher than those obtained with the live vaccine. The association of the live and dead vaccines with the probiotic increased specific anti-PppA antibodies, reaching maximum values in the D-LL + Lc (N) group. The increase in IgA and IgG anti-PppA is of fundamental importance at the lung level, because while IgA prevents pathogen attachment to epithelial cells, during thus reducing colonization, IgG would exert protection at the alveolar level, promoting phagocytosis and preventing local dissemination of the pneumococcus and its passage into blood [35]. We demonstrated that the vaccine-induced humoral immune response was increased in all assessed groups at both the lung and systemic compartments, although the highest levels of specific antibodies were obtained when the vaccine, dead or live, was associated with the probiotic. This was coincident with the increase in IL-4 in the lung compartment, indicating activation of the Th2 cell population, which enhanced the humoral immune response. Recent reports have shown that certain lactobacilli improved the specific antibody response after vaccination against some viral and bacterial pathogens [21,36]. In addition, L.