Gel filtration chromatography was utilized for further purification

n addition, JNK1 2/2 MTEC expressed significantly decreased levels of the antimicrobial peptides S100a8 and Defb4 compared to IL-17A stimulated WT MTEC. Taken together, the data suggest that IL17A signals through JNK1 to induce inflammation and enhance host defense. Since JNK1 was shown to play a role in IL-17A signaling in vitro in epithelial cells, the impact of JNK1 deletion on IL-17A signaling in vivo was investigated. WT and JNK1 2/2 mice were challenged with adenovirus expressing IL-17A for three days. Adenoviral IL-17A induced similar levels of IL-17A protein in the lung; 4088.161069.5 pg/ml in WT mice and 4009.46459.0 pg/ ml in JNK1 2/2 mice. The total numbers of inflammatory cells in the BAL were similar in WT and JNK1 2/2 mice, however, JNK1 2/2 mice had significantly increased macrophage and decreased neutrophil recruitment. In addition to altered cellular infiltrate profiles, JNK1 2/2 mice produced significantly decreased MCP-1 and IFNc compared to WT mice. The adenoviral expression approach utilized introduces the potential caveat of a differential viral response in the WT and JNK1 2/2 mice. To further examine IL-17A signaling in vivo, WT and JNK1 2/2 mice were instilled with recombinant mouse IL-17A for one day. IL-17A induced significantly decreased MCP-1 and G-CSF production, as well as a trend towards lower IP-10 and IFNc, in JNK1 2/2 mice versus WT mice. Furthermore, JNK1 2/2 mice stimulated with IL-17A demonstrated a trend towards decreased antimicrobial peptides S100a8 and S100a9 compared to WT mice. These data show that IL-17A requires JNK1 for inflammatory signaling in vivo. Discussion The results of this study indicate that JNK1 plays a context dependent role in host defense and inflammation. In general, JNK1 was associated with macrophage recruitment in response to each of the three pathogens tested. Furthermore, JNK1 was necessary for induction of MCP-1 and IFNc, two important factors for macrophage function. JNK1 was also implicated in the production of antimicrobial peptides by epithelial cells and in the lung. These data suggest two potential mechanisms by which JNK1 may regulate host defense. In viral pneumonia, JNK1 had a somewhat paradoxical role, as JNK1 2/2 mice had lower viral burden but worsened morbidity and lung histopathology. The mechanism for this did not appear to involve altered mucin gene induction based on the lack of impact on Clca3 mRNA, T cell recruitment, or type I interferon induction. Finally, JNK1 impacted IL-17A signaling in a similar manner to its effects on gram-negative bacterial pneumonia; decreased chemokine and antimicrobial peptide production. These data suggest that IL-17A requires JNK1 signaling which would suggest that JNK1 is required in a number of disease pathologies. The impact of JNK1 in host defense against bacterial pathogens is largely unclear. Little is known about the impact of JNK1 deletion or inhibition in vivo. Pseudomonas aeruginosa induces JNK1 dependent apoptosis of cells via its LY-411575 supplier exotoxin S, E. coli mediated induction of cytokines in HeLa cells was shown to be decreased by JNK1 and Host Defense a JNK inhibitor, and LPS mediated increases of IL-23 was JNK1 dependent. These data support the findings that JNK1 may be important in host defense against gram-negative bacteria. Our data indicate that JNK1 deletion has similar effects on E. coli and IL-17A induced cytokine production. Specifically, IFNc and MCP-1 levels were reduced in JNK1 2/2 mice challenged with bo