PDF(Pubmed)
Abstract：
OBJECTIVE: We aim to describe the liver immune microenvironment by analyzing liver biopsies from patients with chronic HBV infection (CHB). Host immune cell signatures and their corresponding localization were characterized by analyzing the intrahepatic transcriptome in combination with a custom multiplex immunofluorescence panel.
METHODS: Matching FFPE and fresh frozen liver biopsies were collected from immune active patients within the open-label phase IV study GS-US-174-0149. RNA-Seq was conducted on 53 CHB liver biopsies from 46 patients. Twenty-eight of the 53 samples had matched FFPE biopsies and were stained with a 12-plex panel including cell segmentation, immune and viral biomarkers. Corresponding serum samples were screened using the MSD Human V-plex Screen Service to identify peripheral correlates for the immune microenvironment.
RESULTS: Using unsupervised clustering of the transcriptome, we reveal two unique liver immune signatures classified as immune high and immune low based on the quantification of the liver infiltrate gene signatures. Multiplex immunofluorescence analysis demonstrated large periportal lymphoid aggregates in immune high samples consisting of CD4 and CD8 T cells, B cells and macrophages. Differentiation of the high and low immune microenvironments was independent of HBeAg status and peripheral viral antigen levels. In addition, longitudinal analysis indicates that treatment and normalization of ALT correlates with a decrease in liver immune infiltrate and inflammation. Finally, we screened a panel of peripheral biomarkers and identified ICAM-1 and CXCL10 as biomarkers that strongly correlate with these unique immune microenvironments.
CONCLUSIONS: These data provide a description of immune phenotypes in patients with CHB and show that immune responses are downregulated in the liver following nucleotide analogue treatment. This may have important implications for both the safety and efficacy of immune modulator programs aimed at HBV cure.
BACKGROUND: Liver biopsies from patients with chronic hepatitis B were submitted to RNA-Seq and multiplex immunofluorescence and identified two different liver immune microenvironments: immune high and immune low. Immune high patients showed elevated immune pathways, including interferon signaling pathways, and increase presence of immune cells. Longitudinal analysis of biopsies from treatment experienced patients showed that treatment correlates with a marked decrease in inflammation and these findings may have important implications for both safety and efficacy of immune modulator programs for HBV cure.
METHODS: Matching FFPE and fresh frozen liver biopsies were collected from immune active patients within the open-label phase IV study GS-US-174-0149. RNA-Seq was conducted on 53 CHB liver biopsies from 46 patients. Twenty-eight of the 53 samples had matched FFPE biopsies and were stained with a 12-plex panel including cell segmentation, immune and viral biomarkers. Corresponding serum samples were screened using the MSD Human V-plex Screen Service to identify peripheral correlates for the immune microenvironment.
RESULTS: Using unsupervised clustering of the transcriptome, we reveal two unique liver immune signatures classified as immune high and immune low based on the quantification of the liver infiltrate gene signatures. Multiplex immunofluorescence analysis demonstrated large periportal lymphoid aggregates in immune high samples consisting of CD4 and CD8 T cells, B cells and macrophages. Differentiation of the high and low immune microenvironments was independent of HBeAg status and peripheral viral antigen levels. In addition, longitudinal analysis indicates that treatment and normalization of ALT correlates with a decrease in liver immune infiltrate and inflammation. Finally, we screened a panel of peripheral biomarkers and identified ICAM-1 and CXCL10 as biomarkers that strongly correlate with these unique immune microenvironments.
CONCLUSIONS: These data provide a description of immune phenotypes in patients with CHB and show that immune responses are downregulated in the liver following nucleotide analogue treatment. This may have important implications for both the safety and efficacy of immune modulator programs aimed at HBV cure.
BACKGROUND: Liver biopsies from patients with chronic hepatitis B were submitted to RNA-Seq and multiplex immunofluorescence and identified two different liver immune microenvironments: immune high and immune low. Immune high patients showed elevated immune pathways, including interferon signaling pathways, and increase presence of immune cells. Longitudinal analysis of biopsies from treatment experienced patients showed that treatment correlates with a marked decrease in inflammation and these findings may have important implications for both safety and efficacy of immune modulator programs for HBV cure.
摘要：
目的：我们旨在通过分析慢性HBV感染（CHB）患者的肝活检来描述肝脏免疫微环境。通过结合定制的多重免疫荧光面板分析肝内转录组来表征宿主免疫细胞特征及其相应的定位。
方法：在开放标签IV期研究GS-US-174-0149中，从免疫活跃患者中收集匹配的FFPE和新鲜冷冻肝活检。RNA-Seq在46例患者的53例CHB肝活检中进行。53个样本中有28个与FFPE活检相匹配，并用包括细胞分割在内的12组染色，免疫和病毒生物标志物。使用MSD人V-plex筛选服务筛选相应的血清样品,以鉴定免疫微环境的外周相关性。
结果：使用转录组的无监督聚类，我们揭示了两种独特的肝脏免疫特征，根据肝脏浸润基因特征的量化，分为免疫高和免疫低。多重免疫荧光分析显示，由CD4和CD8T细胞组成的高免疫样本中的大型门静脉周围淋巴聚集物，B细胞和巨噬细胞。高低免疫微环境的分化独立于HBeAg状态和外周病毒抗原水平。此外,纵向分析表明，ALT的治疗和正常化与肝脏免疫浸润和炎症的减少相关。最后,我们筛选了一组外周生物标志物,并将ICAM-1和CXCL10鉴定为与这些独特免疫微环境密切相关的生物标志物.
结论：这些数据提供了CHB患者免疫表型的描述，并显示核苷酸类似物治疗后肝脏中的免疫反应下调。这可能对针对HBV治愈的免疫调节剂程序的安全性和有效性都具有重要意义。
背景：来自慢性乙型肝炎患者的肝活检提交RNA-Seq和多重免疫荧光，并确定了两种不同的肝脏免疫微环境：免疫高和免疫低。免疫力高的患者表现出升高的免疫途径，包括干扰素信号通路,并增加免疫细胞的存在。从治疗经验丰富的患者活检的纵向分析表明，治疗与炎症的显着减少相关，这些发现可能对HBV治愈的免疫调节剂程序的安全性和有效性具有重要意义。
方法：在开放标签IV期研究GS-US-174-0149中，从免疫活跃患者中收集匹配的FFPE和新鲜冷冻肝活检。RNA-Seq在46例患者的53例CHB肝活检中进行。53个样本中有28个与FFPE活检相匹配，并用包括细胞分割在内的12组染色，免疫和病毒生物标志物。使用MSD人V-plex筛选服务筛选相应的血清样品,以鉴定免疫微环境的外周相关性。
结果：使用转录组的无监督聚类，我们揭示了两种独特的肝脏免疫特征，根据肝脏浸润基因特征的量化，分为免疫高和免疫低。多重免疫荧光分析显示，由CD4和CD8T细胞组成的高免疫样本中的大型门静脉周围淋巴聚集物，B细胞和巨噬细胞。高低免疫微环境的分化独立于HBeAg状态和外周病毒抗原水平。此外,纵向分析表明，ALT的治疗和正常化与肝脏免疫浸润和炎症的减少相关。最后,我们筛选了一组外周生物标志物,并将ICAM-1和CXCL10鉴定为与这些独特免疫微环境密切相关的生物标志物.
结论：这些数据提供了CHB患者免疫表型的描述，并显示核苷酸类似物治疗后肝脏中的免疫反应下调。这可能对针对HBV治愈的免疫调节剂程序的安全性和有效性都具有重要意义。
背景：来自慢性乙型肝炎患者的肝活检提交RNA-Seq和多重免疫荧光，并确定了两种不同的肝脏免疫微环境：免疫高和免疫低。免疫力高的患者表现出升高的免疫途径，包括干扰素信号通路,并增加免疫细胞的存在。从治疗经验丰富的患者活检的纵向分析表明，治疗与炎症的显着减少相关，这些发现可能对HBV治愈的免疫调节剂程序的安全性和有效性具有重要意义。
