TY - DATA T1 - Supplementary Material for: Intranasal Administration of CpG Induces a Rapid and Transient Cytokine Response Followed by Dendritic and Natural Killer Cell Activation and Recruitment in the Mouse Lung PY - 2017/03/23 AU - Pesce I. AU - Monaci E. AU - Muzzi A. AU - Tritto E. AU - Tavarini S. AU - Nuti S. AU - De Gregorio E. AU - Wack A. UR - https://karger.figshare.com/articles/dataset/Supplementary_Material_for_Intranasal_Administration_of_CpG_Induces_a_Rapid_and_Transient_Cytokine_Response_Followed_by_Dendritic_and_Natural_Killer_Cell_Activation_and_Recruitment_in_the_Mouse_Lung/4781095 DO - 10.6084/m9.figshare.4781095.v1 L4 - https://ndownloader.figshare.com/files/7852807 L4 - https://ndownloader.figshare.com/files/7852816 KW - Adjuvants KW - Lung KW - Dendritic cells KW - Cytokines KW - Mucosal vaccines KW - Natural killer cells N2 - CpG-containing oligodeoxynucleotides are potent mucosal adjuvants and effective as stand-alone treatment of respiratory infections in mice. Although CpG is also used as a type 1 helper immunomodulator in the treatment of asthma and allergic disease, immune modulation following intranasal application has not been fully characterized yet. Using a B-type CpG, we monitored RNA expression profiles, cytokine production and cellular activation in lung tissue and bronchoalveolar lavages ex vivo and cytokine production of purified cell populations in vitro. CpG triggered the upregulation of many transcripts, including interferon response genes and proinflammatory cytokine genes, between 3 h and 4 days. Overlapping subsets of these cytokine proteins were induced in vitro in purified CD11c+ cells, B cells and alveolar macrophages from the lung, thus identifying these cells as direct targets of CpG. While lung B cells strongly respond to CpG in vitro, less activation is found ex vivo, suggesting efficient CpG sequestering or rapid B cell migration after activation. In contrast, a type II alveolar epithelial cell line did not respond to CpG in vitro. We noted selective recruitment of plasmacytoid dendritic cells (DCs) into the lung tissue, and of conventional DCs and natural killer (NK) cells into the lung tissue and bronchoalveolar space. Furthermore, CpG induced activation of intrapulmonary DCs, NK and T cells. We hypothesize that CpG-linked adjuvanticity and clearance of respiratory pathogens are mediated by two major mechanisms: transient induction of the interferon pathway limiting microbial survival and selective recruitment of DCs and NK cells, which allows for better adaptive responses. ER -