With respect to the other cytokines analysed [6], in NALT and NP it was observed that the frequency of IL-2, INF-γ and TNF-α-producing T cells was very low, compared to those producing IL-4, IL-5 and IL-10. They also were increased in immunized mice in relation to control mice (excepting TNF-α-producing cells in NP which did not change). Therefore, although the percentage of T cells that produce IL-2 and IFN-γ represent the lowest values in both groups and in both tissues, these data suggest Torin 1 at least a small number of T cells in NALT and NP produce Th1

cytokines, and their frequency is slightly increased by the effects of Cry1Ac. In previous studies, we have reported that Cry1Ac is highly immunogenic and confers mucosal and systemic adjuvant Neratinib cell line effects when is administered to mice by systemic or mucosal routes [9–12]. In addition, we have observed that Cry1Ac increases protective immunity against experimental N. fowleri meningoencephalitis in mice [14]. Considering that the immune response elicited, following intranasal immunization with Cry1Ac protoxin, had not yet been analysed in the nasal tract, in this work we evaluated specific antibody cell responses, as well as the activation

and cytokine production, in the lymphocyte populations residing at the nasal compartments of the NALT and NP. On the basis of our previous results, and considering the additional advantages that Cry1Ac has over other mucosal adjuvants [10, 17], in that it is non-toxic to vertebrates, and its production costs are low, we had suggested that this protein may be an attractive candidate to improve the efficacy of vaccination against pathogens invading the nasal mucosa. While the outcomes

of present study contribute to explaining the potent immunogenicity of Cry1Ac via i.n. route. Because in both NALT this website and NP lymphocytes from immunized mice we found that: (1) significant specific IgA and IgG cell responses were induced, especially in NP, (2) the proportion of activated lymphocytes was increased and (3) the proportion of T cells spontaneously producing cytokines, especially a Th2 profile of cytokines, was increased. In mice immunized with Cry1Ac, the response found in nasal lymphocytes was as good, or even higher to that attained with CT, which was used as a reference of the most potent mucosal immunogen known. Nevertheless, the immunization scheme used may be not the optimal to achieve the maximal anti-CT responses. Besides, given the different conditions used for each protein, the immune responses achieved are not suitable to compare, because a higher dose of Cry1Ac was used, and because higher doses than 10 μg of CT can not be assayed in mice because of its toxicity.

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