Novel Reporter System Monitoring IL-18 Specific Signaling Can Be Applied to High-Throughput Screening.

Very recently, the immunotherapies against cancer, autoimmune diseases, and infection have been feasible and promising. Thus, we have examined the possibility whether or not human gamma delta T cells can be applied for the novel immunotherapies. We previously established the cells stably maintaining NFkB-driven human secreted embryonic alkaline phosphatase (SEAP) expression. The cells can be used to determine the transcription activity of NFkB with high-standard dynamic range and accuracy. Because IL-18 is a kind of cytokines that enhances cytotoxicity and activity of human gamma delta T cells through NFkB activation, we have focused on the activity and signaling of IL-18. In this study, we modified the previous reporter cell that can determine the transcription activity of NFkB to express two subunits consisted of human IL-18 receptor. The modified cells secreted SEAP in response to treatment with human recombinant IL-18 in a concentration-dependent manner. We also observed the concentration-dependently enhancement of NFkB activity in the cells treated with mouse recombinant IL-18 although the affinity was lower compared to human recombinant IL-18. We also previously established the cells stably expressing and secreting human recombinant IL-18 and then validated whether or not the conditioned medium from the cells activate NFkB transcription activity using this assay. Our university has kept collecting many extracts from over 18,000 marine bacteria in our local sea around Omura bay-fungi, plants for Chinese herbal medicine, and so on-and also have kept gathering synthetic compounds from many Japanese chemists as drug libraries. Finally, in order to identify drugs mimicking IL-18 biological activity or possessing inhibitory effects on IL-18-induced NFkB, we demonstrated drug screening using number of extracts derived from marine bacteria and synthetic compounds.

IL-1 receptor antagonist-deficient mice develop autoimmune arthritis due to intrinsic activation of IL-17-producing CCR2(+)Vγ6(+)γδ T cells.

Interleukin-17 (IL-17)-producing γδ T (γδ17) cells have been implicated in inflammatory diseases, but the underlying pathogenic mechanisms remain unclear. Here, we show that both CD4(+) and γδ17 cells are required for the development of autoimmune arthritis in IL-1 receptor antagonist (IL-1Ra)-deficient mice. Specifically, activated CD4(+) T cells direct γδ T-cell infiltration by inducing CCL2 expression in joints. Furthermore, IL-17 reporter mice reveal that the Vγ6(+) subset of CCR2(+) γδ T cells preferentially produces IL-17 in inflamed joints. Importantly, because IL-1Ra normally suppresses IL-1R expression on γδ T cells, IL-1Ra-deficient mice exhibit elevated IL-1R expression on Vγ6(+) cells, which play a critical role in inducing them to produce IL-17. Our findings demonstrate a pathogenic mechanism in which adaptive and innate immunity induce an autoimmune disease in a coordinated manner.

Improvement in the bioavailability of poorly absorbed glycyrrhizin via various non-vascular administration routes in rats.

The purpose of this study was to examine the improvement of the bioavailability of glycyrrhizin (GL) via extra-vascular, i.e. oral, rectal, and nasal routes with or without the aid of an absorption enhancer in place of the vascular intravenous route in rats. Pharmacokinetic behavior following administration via vascular routes, i.e. the intravenous and portal-venous routes was examined in rats. The area under the plasma concentration-time curve (AUC) after administration of GL via the portal vein was decreased slightly, suggesting that the first elimination of GL in the liver may be one of the factors contributing to the low bioavailability after administration via the oral route. When GL was administered orally as a solution (30 mg/kg), the plasma concentration of GL was extremely low. However, after rectal or nasal administration of GL solution (30 mg/kg) with or without sodium caprate, the mean AUC value was remarkably increased compared with oral administration. In particular, the absolute bioavailability of GL after nasal administration was estimated to be approximately 20%, which was approximately 80-fold greater compared with after oral administration despite of the absence of an enhancer. Furthermore, the fatty acids co-administered orally with GL produced an increase in GL absorption in the following order: sodium caprate>sodiumlaurate>sodiumcaprylate>sodium oleate. These results indicate that the rectum and nasal cavity are useful administration routes for systemic delivery of GL. It was also found that the fatty acids were enhancers for the absorption of GL.