While earlier treatments, such as cytoreductive chemo and interferon therapies

jejuni wild type strain, This finding suggests that the invasion phenotype of the ciaD mutant is due to a lack in the initiation of cellular signaling events specific to invasion, and not from the failure to induce the secretion of IL 8 from host cells. We also confirmed that Caco 2 cells are purchaseABT-888 responsive to IL 8, using immunoblot analysis to quantify phospho Akt; Akt is a downstream target of the CXCR1 2 receptors and is activated by IL 8, Together, these experiments revealed that C. jejuni must activate components of the MAP kinase signaling pathway for both cellular invasion and the secretion of IL 8, and that CiaD contributes to this activation. CiaD activates of the MAP kinase signaling pathway Based on the presence of the Mitogen activated protein kinase docking motif in CiaD, experiments were performed to determine if C.

jejuni activated the MAP kinase signaling pathway in a CiaD dependent manner. Prior to performing these experiments, we determined that the C. jejuni wild type strain induces significantly more Afatinib HER2 阻害剤 IL 8 than the ciaD mutant at 4 hr post inoculation of INT 407 cells, Based on the kinetics of IL 8 secretion, we performed experiments to identify the cellular signaling pathways that are activated by C. jejuni at 3 hr post inoculation. The use of a MAP kinase phosphor array screen revealed that the C. jejuni ciaD mutant did not activate Erk and p38 to the same extent as inoculation of the INT 407 cells with the wild type strain, These results were confirmed by immunoblot analysis, The partial activation of Erk by the ciaD mutant is consistent with the fact that Erk is also partially activated in response to host cell binding mediated by FlpA, In contrast to the C.

jejuni ciaD and flgBC mutants, the C. jejuni wild type strain and purchase AG-1478 ciaD mutant harboring a wild type copy of the ciaD gene resulted in the activation of Erk and p38 as judged by an increase in band intensity of the phos phorylated form of the protein. Even though the C. jejuni wild type strain, ciaD mutant, and complemented ciaD isolate resulted in greater activation of NF κB compared to the flgBC mutant, significant differences were not observed in the amount of NF κB activation between the isolates, This finding suggested that a protein or other bacterial component other than CiaD is re sponsible for the activation of NF κB.

Taken together, our results indicate that CiaD participates in the activation of the MAP kinase signaling molecules Erk and p38. The MAP Kinase docking motif of CiaD is required for IL 8 secretion and host cell invasion Mutational analysis was used to determine whether the putative eukaryotic domains of CiaD are functional. Two C. jejuni ciaD mutants were generated; the MAP kinase docking motif was deleted in one mutant and the proline directed phosphorylation motif was changed to an alanine P) in the other mutant, Immunoblot analysis revealed that both CiaD recom binant proteins were synthesized in the ciaD mutant, Importantly, all of the isolates were motile, Ex periments were then performed to evaluate the ability of the CiaD MKD mutant and CiaD P mutant to induce IL 8 secretion and to invade human INT 407 epithelial cells.