| FICZ Exposure and Viral Infection in Mice
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Author:
Date:
2017-01-05
[Abstract] The aryl hydrocarbon receptor (AHR) is known as a sensor for dioxins that mediates their toxicity, and also has important biophysiological roles such as circadian rhythms, cell differentiation and immune responses. 6-formylindolo(3,2-b)carbazole (FICZ), which is derived through the metabolism of L-tryptophan by ultraviolet B irradiation, is one of putative physiological ligands for AHR (Smirnova et al., 2016). It has recently been shown that endogenously-activated AHR signaling modulates innate immune response during viral infection (Yamada et al., 2016). This section describes how to treat mice with FICZ and to infect them with virus.
[摘要] 芳烃受体（AHR）被称为二恶英介导其毒性的传感器，也具有重要的生物生理学作用，如昼夜节律，细胞分化和免疫应答。通过紫外线B照射通过L-色氨酸的代谢衍生的6-甲酰基吲哚（3,2-b）咔唑（FICZ）是AHR的推定生理配体之一（Smirnova等人）。 ，2016）。最近已经显示内源性激活的AHR信号调节病毒感染期间的先天免疫应答（Yamada等人，2016）。本节介绍如何用FICZ治疗小鼠并用病毒感染。
背景 迄今为止，AHR的作用主要是在二恶英治疗实验的基础上进行了调查。另一方面，已经显示AHR介导的信号传导是由内源色氨酸代谢物（FICZ，Kynurenine，等等），血红素代谢物（胆红素等）激活的。 ，和类二十烷酸（Lipoxin A等）。特别地，已经证明FICZ是AHR的生理高亲和力配体，许多积累的报道显示FICZ参与各种基本生物学过程，包括对紫外线的适应性反应，免疫应答，基因组不稳定性和干细胞的体内平衡。最近，Yamada等人。 （2016）证明其在先天免疫应答中的作用：体内FICZ治疗抑制响应于病毒感染的I型干扰素（IFN）产生并促进小鼠血清中病毒滴度的水平。
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| In vitro Treatment of Mouse and Human Cells with Endogenous Ligands for Activation of the Aryl Hydrocarbon Receptor
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Author:
Date:
2017-01-05
[Abstract] Activation of the aryl hydrocarbon receptor (AHR) by endogenous ligands has been implicated in a variety of physiological processes such as cell cycle regulation, cell differentiation and immune responses. It is reported that tryptophan metabolites, such as kynurenine (Kyn) and 6-formylindolo(3,2-b)carbazole (FICZ), are endogenous ligands for AHR (Stockinger et al., 2014). This protocol is designed for treatment with Kyn or FICZ in mouse embryonic fibroblasts (MEFs) or primary peripheral monocytes.
[摘要] 通过内源性配体活化芳基烃受体（AHR）已涉及多种生理过程，如细胞周期调控，细胞分化和免疫应答。据报道，色氨酸代谢物，如犬尿胆碱（Kyn）和6-甲基吲哚（3,2-b）咔唑（FICZ）是AHR的内源性配体（Stockinger等人，2014）。该方案设计用于在小鼠胚胎成纤维细胞（MEF）或初级周边单核细胞中用Kyn或FICZ进行治疗。
背景 色氨酸代谢物如Kyn和FICZ是生理条件下AHR的内源性配体。 Kyn由色氨酸-2,3-双加氧酶（TDO）和/或吲哚胺-2,3-双加氧酶1和2（IDO1 / 2）产生，并有助于抑制抗肿瘤反应和恶性进展（Stockinger等人，2014）。 ...
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| HBV Infection in Human Hepatocytes and Quantification of Encapsidated HBV DNA
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Author:
Date:
2016-01-20
[Abstract] Human hepatic cancer cell lines such as HepG2, Huh7, and HLE cannot get infected with Hepatitis B virus (HBV) due to lack of an HBV receptor(s). Transfection with HBV genome has so far been referred as a tool to mimic HBV infection. However, since sodium taurocholate cotransporting polypeptide (NTCP) was identified as a functional receptor for HBV (Yan et al., 2012), hepatocyte cell lines that were stably transfected with a plasmid for NTCP expression have been used for HBV infection. This protocol is designed for infection with HBV in human hepatocyte cell line HepG2 expressing NTCP (HepG2-hNTCP-C4 cells; Iwamoto et al., 2014) or primary human hepatocytes (PHHs). In this section, we also describe one of the methods for the assessment of HBV infection: Quantification of ...
[摘要] 人肝癌细胞系如HepG2，Huh7和HLE由于缺乏HBV受体而不能感染乙型肝炎病毒（HBV）。 HBV基因组的转染迄今为止被称为模拟HBV感染的工具。 然而，由于牛磺胆酸钠共转运多肽（NTCP）被鉴定为HBV的功能性受体（Yan等人，2012），已经使用用用于NTCP表达的质粒稳定转染的肝细胞细胞系 为HBV感染。 该方案设计用于在表达人类肝细胞细胞系HepG2的表达NTCP（HepG2-hNTCP-C4细胞; Iwamoto等人，2014）或原代人肝细胞（PHH）的HBV感染。 在本节中，我们还描述了用于评估HBV感染的方法之一：细胞内衣壳化的HBV DNA的定量。
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