Background The impact of nucleos(t)ide analogues on intrahepatic viral burden and immune microenvironment in patients with chronic hepatitis B (CHB) is not clear. Objective We aimed to characterise the effects of tenofovir disoproxil fumarate (TDF) on intrahepatic viral burden and the liver immune microenvironment in patients with CHB. Design Core liver biopsies were collected at baseline and year 3 from patients with CHB with minimally raised serum alanine aminotransferase in a double-blind placebo-controlled trial ([NCT01522625][1]). Paired biopsies were analysed by RNA-sequencing (n=119 pairs), a custom multiplex immunofluorescence assay (n=30 pairs), and HBV-targeted long-read DNA sequencing (n=49 pairs). Results Both non-integrated and integrated HBV DNA were present in all patients at baseline, with >65% having interchromosomal translocations. Treatment significantly reduced the frequency of HBV core+ hepatocytes and intrahepatic (integrated and non-integrated) HBV DNA, but had no effect on HBsAg+ hepatocytes. Clonally expanded integrations were enriched for HBsAg coding regions and showed dysregulation of nearby genes. At baseline, there was significant enrichment of intrahepatic CD8+ T cell proximity to HBV core+ hepatocytes, but not to HBsAg+ cells. The densities of T cells and B cells were significantly reduced by TDF. Transcriptomic analyses found TDF induced widespread downregulation of immune-related genes including inhibitory and regulatory genes. Conclusion TDF significantly reduced intrahepatic integrated and non-integrated HBV DNA, exerting disparate effects on HBV core+ and HBsAg+ cells and on different immune cell subsets. Our data suggest there may be differential cytotoxic T cell-mediated killing of HBV core+ versus HBsAg+ hepatocytes, providing insights for HBV cure strategies. Data are available upon reasonable request. Data collected for this study, including deidentified individual participant data and data dictionaries defining fields in the datasets, will be made available on request to with qualified external researchers based on submitted curriculum vitae and reflecting non conflict of interest as well as laws and regulations regarding patient privacy. Approval of such requests is dependent on the nature of the request, the merit of the research proposed, the availability of the data and the intended use of the data. Data requests should be sent to the corresponding author at holdenhsu@gmail.com. [1]: /lookup/external-ref?link_type=CLINTRIALGOV&access_num=NCT01522625&atom=%2Fgutjnl%2Fearly%2F2024%2F10%2F08%2Fgutjnl-2024-332526.atom

中文翻译：

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富马酸替诺福韦二吡呋酯对慢性乙型肝炎患者肝内病毒载量和肝脏免疫微环境的影响


背景 核苷（酸）类似物对慢性乙型肝炎 （CHB） 患者肝内病毒载量和免疫微环境的影响尚不清楚。目的 我们旨在表征富马酸替诺福韦二吡呋酯 （TDF） 对 CHB 患者肝内病毒负荷和肝脏免疫微环境的影响。设计 在一项双盲安慰剂对照试验中，在基线和第 3 年收集血清丙氨酸氨基转移酶轻度升高的 CHB 患者的核心肝活检 （[NCT01522625][1]）。通过 RNA 测序 （n=119 对） 、定制多重免疫荧光测定 （n=30 对） 和 HBV 靶向长读长 DNA 测序 （n=49 对） 分析配对活检。结果 基线时所有患者均存在非整合和整合的 HBV DNA，其中 >65% 存在染色体间易位。治疗显著降低了 HBV 核心 + 肝细胞和肝内 （整合和非整合） HBV DNA 的频率，但对 HBsAg + 肝细胞没有影响。克隆扩增的整合富集了 HBsAg 编码区，并显示附近基因的失调。基线时，肝内 CD8 + T 细胞显著富集靠近 HBV 核心 + 肝细胞，但不富集 HBsAg + 细胞。TDF 显著降低了 T 细胞和 B 细胞的密度。转录组学分析发现 TDF 诱导免疫相关基因（包括抑制基因和调节基因）的广泛下调。结论 TDF 显著降低肝内整合和非整合 HBV DNA，对 HBV 核心 + 和 HBsAg+ 细胞以及不同的免疫细胞亚群产生不同的影响。 我们的数据表明，细胞毒性 T 细胞介导的 HBV 核心 + 杀伤与 HBsAg+ 肝细胞可能存在差异，为 HBV 治愈策略提供了见解。数据可根据合理要求提供。为本研究收集的数据，包括去识别化的个人参与者数据和定义数据集中字段的数据字典，将根据提交的简历并反映非利益冲突以及有关患者隐私的法律法规，应要求提供给合格的外部研究人员。此类请求的批准取决于请求的性质、拟议研究的优点、数据的可用性以及数据的预期用途。数据请求应通过 holdenhsu@gmail.com 发送给通讯作者。[1]： /lookup/external-ref？link_type=CLINTRIALGOV&access_num=NCT01522625&atom=%2Fgutjnl%2Fearly%2F2024%2F10%2F08%2Fgutjnl-2024-332526.原子