Effects of tenofovir disoproxil fumarate on intrahepatic viral burden and liver immune microenvironment in patients with chronic hepatitis B.

We aimed to characterise the effects of tenofovir disoproxil fumarate (TDF) on intrahepatic viral burden and the liver immune microenvironment in patients with CHB.

我们旨在描述替诺福韦二丙酸酯(TDF)对CHB患者肝内病毒负荷和肝脏免疫微环境的影响。

Methods

Core liver biopsies were collected at baseline and year 3 from patients with CHB with minimally raised serum alanine aminotransferase in a double-blindplacebo-controlledtrial (NCT01522625).

在一项双盲安慰剂对照试验（NCT01522625）中，从基线和第3年收集了血清丙氨酸氨基转移酶仅轻微升高的慢性乙型肝炎患者的肝核心活检样本。

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).

通过RNA测序（n=119对）、定制的多重免疫荧光检测（n=30对）和针对HBV的长读取DNA测序（n=49对）分析了配对的活检样本。

Results

Both non-integrated and integrated HBV DNA were present in all patients at baseline, with >65% having interchromosomal translocations.

在所有患者中，基线时均存在非整合和整合的乙型肝炎病毒（HBV）DNA，超过65%的患者有染色体间易位。

Treatment significantly reduced the frequency of HBV core+ hepatocytes and intrahepatic (integrated and non-integrated) HBV DNA, but had no effect on HBsAg+ hepatocytes.

治疗显著降低了乙型肝炎病毒核心蛋白阳性肝细胞和肝内（整合和非整合）乙型肝炎病毒DNA的频率，但对乙型肝炎表面抗原阳性肝细胞无影响。

Clonally expanded integrations were enriched for HBsAg coding regions and showed dysregulation of nearby genes.

克隆性扩增的整合位点在乙型肝炎表面抗原（HBsAg）编码区富集，并显示出附近基因的失调。

At baseline, there was significant enrichment of intrahepatic CD8+ T cell proximity to HBV core+ hepatocytes, but not to HBsAg+ cells.

在基线时，肝内CD8+ T细胞与乙型肝炎病毒核心抗原（HBV core）阳性肝细胞的接近性显著增加，但与乙型肝炎表面抗原（HBsAg）阳性细胞无此现象。

The densities of T cells and B cells were significantly reduced by TDF.

TDF显著降低了T细胞和B细胞的密度。

Transcriptomic analyses found TDF induced widespread downregulation of immune-related genes including inhibitory and regulatory genes.

转录组分析发现，TDF诱导了包括抑制性和调节性基因在内的免疫相关基因的广泛下调。

Conclusions

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.

TDF显著降低了肝内整合和非整合HBV DNA，对HBV核心+和HBsAg+细胞以及不同免疫细胞亚群产生了不同的影响。

Our data suggest there may be differential cytotoxic T cell-mediated killing of HBV core+ versus HBsAg+ hepatocytes, providing insights for HBV cure strategies.

我们的数据表明，HBV核心+与HBsAg+肝细胞之间可能存在不同的细胞毒性T细胞介导的杀伤作用，为HBV治愈策略提供了见解。