Homo sapiens Raw sequence reads. Homo sapiens

SummaryA population of CD8+ T cells expressing the C–X–C chemokine receptor type 5 (CXCR5) display more potent efficacy on viral replication than the CXCR5− counterpart in mice with chronic lymphocytic choriomeningitis virus (LCMV) infection, which suggest that this subset may exist in a reduced state of 'exhaustion' and maintain suboptimal but critical function. However, it remains unknown about the transcriptomes of this subset in patients with chronic hepatitis B virus (HBV) infection. Herein, we determined the transcriptomes of splenic CXCR5+CD8+ T cells and CXCR5−CD8+ T cells in patients with chronic HBV infection.Overall designSplenic lymphocytes were obtained from 3 patients who underwent splenectomy due to HBV-related liver cirrhosis induced hypersplenism, then the CXCR5+CD8+ T cells and CXCR5-CD8+ T cells were sorted. Total RNA was acquired and libraries for RNA-seq were prepared using the BGISEQ-500 platform. KEGG pathway analysis showed that, although the two subsets shared a mass of molecular characteristics, we found over one thousand transcripts that were significantly downregulated or upregulated (two fold or more) in CXCR5+CD8+ T cells relative to their expression in the CXCR5− subset. It is worth noting the CXCR5+ subset showed lower expression of transcripts encoding effector molecules involved in cytotoxicity, including granzyme and perforin, while had higher expression of transcripts encoding TNF, MX1, and ISG15). Conclusions: CXCR5+CD8+ T cells displayed as a distinct subset that differed transcriptionally from the CXCR5−CD8+ T cells in chronic HBV infection.

研究概要：在慢性淋巴细胞性脉络丛脑膜炎病毒（Lymphocytic choriomeningitis virus, LCMV）感染小鼠模型中，相较于CXCR5阴性（CXCR5−）的对应细胞群，表达C-X-C趋化因子受体5型（C-X-C chemokine receptor type 5, CXCR5）的CD8阳性T细胞群对病毒复制展现出更强的抑制效能，这提示该细胞亚群可能处于“耗竭”程度较轻的状态，并维持着虽非最优但至关重要的功能。然而，慢性乙型肝炎病毒（Hepatitis B virus, HBV）感染患者体内该亚群的转录组特征仍未明确。为此，本研究解析了慢性HBV感染患者脾脏中CXCR5阳性CD8阳性T细胞与CXCR5阴性CD8阳性T细胞的转录组。
实验设计：本研究从3例因HBV相关性肝硬化引发脾功能亢进而接受脾切除术的患者体内获取脾脏淋巴细胞，随后分选出CXCR5阳性CD8阳性T细胞与CXCR5阴性CD8阳性T细胞。提取总RNA并构建RNA测序文库，采用BGISEQ-500平台完成测序。京都基因与基因组百科全书（Kyoto Encyclopedia of Genes and Genomes, KEGG）通路分析结果显示，尽管两个细胞亚群共享大量分子特征，但相较于CXCR5阴性亚群，CXCR5阳性亚群中存在超过1000个差异表达显著（上调或下调幅度≥2倍）的转录本。值得注意的是，CXCR5阳性亚群中参与细胞毒性效应的分子编码转录本（如颗粒酶（granzyme）与穿孔素（perforin））的表达水平更低，而肿瘤坏死因子（Tumor Necrosis Factor, TNF）、MX1以及ISG15的编码转录本表达水平更高。
结论：在慢性HBV感染患者体内，CXCR5阳性CD8阳性T细胞是一类转录组特征与CXCR5阴性CD8阳性T细胞显著不同的独特细胞亚群。