Agonistic Activation of Cytosolic DNA Sensing Receptors in Woodchuck Hepatocyte Cultures and Liver for Inducing Antiviral Effects.
Agonistic Activation of Cytosolic DNA Sensing Receptors in Woodchuck Hepatocyte Cultures and Liver for Inducing Antiviral Effects.
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DOI：
10.3389/fimmu.2021.745802
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发表时间：
2021
影响因子：
7.3
通讯作者：
Menne S
中科院分区：
文献类型：
--
作者：
Suresh M;Li B;Huang X;Korolowicz KE;Murreddu MG;Gudima SO;Menne S
近年来，随着越来越多的针对不同宿主模式识别受体（PRR）的化合物的出现，用于治疗慢性乙型肝炎（CHB）的免疫调节越来越受到关注。这些激动分子激活受体信号通路并触发先天免疫反应，最终形成控制乙型肝炎病毒 (HBV) 慢性感染的适应性免疫。虽然病毒感染过程中 PRR 对 HBV 核酸的明确识别仍需阐明，但临床前和临床上已探索了几种病毒 RNA 传感受体，包括 Toll 样受体 7/8/9 和视黄酸诱导基因 I 样受体，作为可能的抗 HBV 靶点。病毒 DNA 传感受体的抗病毒潜力研究较少。在本研究中，用HSV-60或聚(dA:dT)激动剂处理CHB动物产生的原代土拨鼠肝细胞，导致干扰素γ诱导蛋白16 (IFI16)或Z-DNA结合蛋白1 (ZBP1/DAI)表达增加，而黑色素瘤2 (AIM2)受体及其各自的衔接分子和效应细胞因子不表达。细胞质 DNA 传感受体通路激活与这些细胞中土拨鼠肝炎病毒 (WHV) 复制和分泌的下降相关。 HSV-60 和聚 (dA:dT) 的联合治疗比任一激动剂的单一治疗取得了更好的抗病毒效果，并且与效应细胞因子表达增加相关。然而，通过外源提供额外的 I 型干扰素 (IFN) 无法进一步增强抗病毒作用，这表明这些受体在激动后产生的效应细胞因子达到饱和水平。在未感染 WHV 的土拨鼠中，通过肝脏靶向递送施用的单剂量聚（dA:dT）耐受性良好，并诱导 ZBP1/DAI 和 AIM2 受体及其效应细胞因子、IFN-β 以及白细胞介素 1β 和 18 的肝内表达。受体激动作用还导致外周血细胞的 IFN-γ 分泌增加。总而言之，体外激活 IFI16、ZBP1/DAI 和 AIM2 受体途径后对 WHV 复制和分泌的影响表明，靶向多个胞质 DNA 受体具有抗病毒益处。此外，肝脏中 ZBP1/DAI 和 AIM2 受体途径的体内激活表明，激动剂递送方法用于未来评估患有 CHB 的土拨鼠的 HBV 治疗效果的可行性。
Immune modulation for the treatment of chronic hepatitis B (CHB) has gained more traction in recent years, with an increasing number of compounds designed for targeting different host pattern recognition receptors (PRRs). These agonistic molecules activate the receptor signaling pathway and trigger an innate immune response that will eventually shape the adaptive immunity for control of chronic infection with hepatitis B virus (HBV). While definitive recognition of HBV nucleic acids by PRRs during viral infection still needs to be elucidated, several viral RNA sensing receptors, including toll-like receptors 7/8/9 and retinoic acid inducible gene-I-like receptors, are explored preclinically and clinically as possible anti-HBV targets. The antiviral potential of viral DNA sensing receptors is less investigated. In the present study, treatment of primary woodchuck hepatocytes generated from animals with CHB with HSV-60 or poly(dA:dT) agonists resulted in increased expression of interferon-gamma inducible protein 16 (IFI16) or Z-DNA-binding protein 1 (ZBP1/DAI) and absent in melanoma 2 (AIM2) receptors and their respective adaptor molecules and effector cytokines. Cytosolic DNA sensing receptor pathway activation correlated with a decline in woodchuck hepatitis virus (WHV) replication and secretion in these cells. Combination treatment with HSV-60 and poly(dA:dT) achieved a superior antiviral effect over monotreatment with either agonist that was associated with an increased expression of effector cytokines. The antiviral effect, however, could not be enhanced further by providing additional type-I interferons (IFNs) exogenously, indicating a saturated level of effector cytokines produced by these receptors following agonism. In WHV-uninfected woodchucks, a single poly(dA:dT) dose administered via liver-targeted delivery was well-tolerated and induced the intrahepatic expression of ZBP1/DAI and AIM2 receptors and their effector cytokines, IFN-β and interleukins 1β and 18. Receptor agonism also resulted in increased IFN-γ secretion of peripheral blood cells. Altogether, the effect on WHV replication and secretion following in vitro activation of IFI16, ZBP1/DAI, and AIM2 receptor pathways suggested an antiviral benefit of targeting more than one cytosolic DNA receptor. In addition, the in vivo activation of ZBP1/DAI and AIM2 receptor pathways in liver indicated the feasibility of the agonist delivery approach for future evaluation of therapeutic efficacy against HBV in woodchucks with CHB.