Fang Zhong, Wang Cong, Huang Chen-Lu, Tan Dan, Peng Xiu-Hua, Bai Jin-Jin, Yuan Zheng-Hong, Yu Xiao-Yu, Ren Guang-Xu
Liver Cancer Institute of Zhongshan Hospital and Key Laboratory of Carcinogenesis and Cancer Invasion (Ministry of Education), Fudan University, Shanghai, China.
Key Laboratory of Medical Molecular Virology (MOE/NHC/CAMS), Research Unit of Cure of Chronic Hepatitis B Virus Infection (CAMS), Shanghai Frontiers Science Center of Pathogenic Microbes and Infection, School of Basic Medical Sciences, Shanghai Medical College, Fudan University, Shanghai, China.
J Med Virol. 2025 Apr;97(4):e70303. doi: 10.1002/jmv.70303.
Hepatitis B virus (HBV) infection is a worldwide health problem. Both CD4+ and CD8 + T cells play crucial roles in HBV clearance from acute patients. Nevertheless, an extrathymic CD4 and CD8 double positive T (DPT) cell subset have been reported earlier, the function of these cells in HBV infection is still poorly understood. Herein, peripheral blood mononuclear cells were collected from hepatitis B patients. HBV model mice were established via hydrodynamic injection (HDI) of pAAV-HBV1.2 plasmid. T cells subsets were analyzed with flow cytometry. We found that in acute HBV infection extrathymic DPT cells were significantly increased in acute patients and HDI-based HBV model mice. Unlike thymic DPT cells, these extrathymic DPT cells activated with a CD44 + CD62L+ central memory phenotype. Furthermore, in vitro cultured DPT cells showed the capability to rapidly proliferate and produce multi cytokines after stimulation with HBV peptides. The performance of adoptive transfer depicted that DPT cells were able to migrate into the liver. Immunohistochemistry data from liver biopsies of patients showed that DPT cells were more prone to detection in acute tissue. Purified DPT cells could efficiently kill HBV peptide-loaded hepatocytes in a cytotoxicity assay, and the frequency of DPT cells were reversely correlated with HBV clearance in model mice. Importantly, the transferred DPT cells accelerated the clearance of HBV in mice. Collectively, our study revealed that extrathymic DPT cells are an important immune subset, contributing to viral clearance during HBV infection, which may benefit cure of chronic hepatitis B.
乙型肝炎病毒(HBV)感染是一个全球性的健康问题。CD4⁺和CD8⁺T细胞在急性患者清除HBV的过程中都发挥着关键作用。然而,此前已有胸腺外CD4和CD8双阳性T(DPT)细胞亚群的报道,这些细胞在HBV感染中的功能仍知之甚少。在此,我们从乙型肝炎患者中收集外周血单个核细胞。通过水动力注射(HDI)pAAV - HBV1.2质粒建立HBV模型小鼠。用流式细胞术分析T细胞亚群。我们发现,在急性HBV感染中,急性患者和基于HDI的HBV模型小鼠的胸腺外DPT细胞显著增加。与胸腺DPT细胞不同,这些胸腺外DPT细胞以CD44⁺CD62L⁺中央记忆表型被激活。此外,体外培养的DPT细胞在受到HBV肽刺激后显示出快速增殖和产生多种细胞因子的能力。过继转移实验表明DPT细胞能够迁移到肝脏。患者肝脏活检的免疫组织化学数据显示,在急性组织中更容易检测到DPT细胞。在细胞毒性试验中,纯化的DPT细胞能够有效杀伤负载HBV肽的肝细胞,并且DPT细胞的频率与模型小鼠中HBV的清除呈负相关。重要的是,转移的DPT细胞加速了小鼠体内HBV的清除。总的来说,我们的研究表明胸腺外DPT细胞是一个重要的免疫亚群,有助于在HBV感染期间清除病毒,这可能有益于慢性乙型肝炎的治疗。