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AdjuvantImmunization with Antigen and Mucosal Immune Responses after Mucosal NK Cells Influence Both Innate and Adaptive

NK cells were found to be recruited in a temporally controlled manner to the nasal-associated lymphoid tissue and the cervical lymph nodes of mice after intranasal immunization with Ag85B-early secreted antigenic target 6kDa from Mycobacterium tuberculosis mixed with Escherichia coli heat-labile toxin as adjuvant. These NK cells were activated and secreted a diverse range of cytokines and other immunomodulators. Using Ab depletion targeting anti-asialo GM1, we found evidence for altered trafﬁcking, impaired activation, and cytokine secretion of dendritic cells, macrophages, and neutrophils in immunized NK cell-depleted mice compared with control animals. Analysis of Ag-speciﬁc immune responses revealed an attenuated Ab and cytokine response in immunized NK cell-depleted animals. Systemic administration of rIL-6 but not rIFN- g signiﬁcantly restored immune responses in mice depleted of NK cells. In conclusion, cytokine production, particularly IL-6, via NK cells and NK cell-activated immune populations plays an important role in the establishment of local innate immune responses and the consequent development of adaptive immunity after mucosal immunization. The Journal of Immunology , 2010, 184: 4327–4337. The level of HRP associated with each well was then determined using Sigma Fast o -phenylenediamine dihydrochloride (50 m l per well; Sigma-Aldrich) colorimetric substrate. The reaction was stopped after 15 min by the addition of 2.5 M H 2 SO 4 . The optical density (OD) 490 was determined, and the titer was expressed as the reciprocal of the dilution giving an OD of 0.2.