Education and Research Center for Pharmaceutical Sciences, Osaka University of Pharmaceutical Sciences, 4-20-1 Nasahara, Takatsuki, Osaka 569-1094, Japan.
Division of Immune Regulation, Institute for Genome Research, Tokushima University, Tokushima-shi, Tokushima 770-8505, Japan.
Mar Drugs. 2020 Jan 17;18(1):60. doi: 10.3390/md18010060.
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.
最近，癌症、自身免疫性疾病和感染的免疫疗法已经可行并具有广阔前景。因此，我们研究了人类 γδ T 细胞是否可用于新型免疫疗法。我们先前建立了稳定表达 NFkB 驱动的人分泌型碱性磷酸酶（SEAP）表达的细胞。这些细胞可用于以高精度和高动态范围来确定 NFkB 的转录活性。因为 IL-18 是一种细胞因子，可通过 NFkB 激活来增强人 γδ T 细胞的细胞毒性和活性，所以我们专注于 IL-18 的活性和信号转导。在这项研究中，我们修改了先前可确定 NFkB 转录活性的报告细胞，使其表达由人 IL-18 受体的两个亚基组成。修饰后的细胞可响应人重组 IL-18 的处理而分泌 SEAP，呈浓度依赖性。我们还观察到，在用鼠重组 IL-18 处理的细胞中，NFkB 活性呈浓度依赖性增强，尽管与重组人 IL-18 相比亲和力较低。我们还先前建立了稳定表达和分泌人重组 IL-18 的细胞，然后使用该测定法验证细胞条件培养基是否激活 NFkB 转录活性。我们大学一直在收集来自我们位于大村湾周围的本地海域的 18000 多种海洋细菌的许多提取物——真菌、中药用植物等——并从许多日本化学家那里收集合成化合物作为药物库。最后，为了鉴定模拟 IL-18 生物学活性或对 IL-18 诱导的 NFkB 具有抑制作用的药物，我们使用源自海洋细菌的提取物和合成化合物进行了药物筛选。
