临床肝胆病杂志2025，Vol.41Issue(4)：628-636,9.DOI:10.12449/JCH250406
聚乙二醇干扰素-α-2b对HBeAg阴性慢性HBV感染者HBV特异性CD8+T淋巴细胞杀伤功能的影响
Effect of pegylated interferon-α-2b therapy on cytotoxicity of virus-specific CD8+T cells in HBeAg-negative patients with chronic hepatitis B virus infection
摘要
Abstract
Objective To investigate the change in the activity of hepatitis B virus(HBV)-specific CD8+T cells after pegylated interferon-α-2b(PEG-IFN-α-2b)therapy in HBeAg-negative patients with chronic HBV infection.Methods A total of 53 HBeAg-negative patients with chronic HBV infection who attended The First Affiliated Hospital of Xinxiang Medical University and Tangdu Hospital of Air Force Mdical University from April 2020 to June 2022 were enrolled and treated with PEG-IFN-α-2b(180 μg/week,subcutaneous injection)antiviral therapy.The study endpoint was HBsAg clearance(course of treatment<48 weeks)or 48 weeks(course of treatment≥48 weeks).Peripheral blood mononuclear cells were isolated at baseline and study endpoint,and peripheral blood T cell counts were measured.Enzyme-linked immunospot assay was used to measure the frequency of HBV-specific CD8+T cells secreting perforin,granzyme B,and interferon-γ.A total of 17 HLA-A*02-restricted patients were selected,and CD8+T cells were purified to establish direct-and indirect-contact co-culture systems for HBV-specific CD8+T cells and HepG2.2.15 cells.The level of lactate dehydrogenase in supernatant was measured to calculate the mortality rate of HepG2.2.15 cells,and the levels of HBV DNA,cytotoxic molecules,and cytokines in supernatant were also measured.Flow cytometry was used to measure the expression of apoptosis ligands,and the cytotoxicity of HBV-specific CD8+T cells was evaluated.The independent samples t-test or the paired t-test was used for comparison of normally distributed continuous data between two groups,and the Mann-Whitney U test or the Wilcoxon rank-sum test was used for comparison of non-normally distributed continuous data between two groups.Results The HBsAg clearance rate at study endpoint was 30.19%(16/53).There were no significant differences in peripheral blood T cell counts(CD3+,CD4+,and CD8+T cells)between baseline and study endpoint(P>0.05).At study endpoint,there was a significant increase in the frequency of HBV-specific CD8+T cells secreting perforin,granzyme B,and interferon-γ(U=177.50,t=11.90,U=186.50,all P<0.001),and the patients with HBsAg clearance had a significantly higher frequency of such HBV-specific CD8+T cells than those without HBsAg clearance(U=120.50,t=2.73,U=121.50,all P<0.01).In the direct-and indirect-contact co-culture systems at study endpoint,HBV-specific CD8+T cells induced a significant reduction in HBV DNA in the supernatant of HepG2.2.15 cells(all P<0.001)and significant increases in the secretion of interferon-γ and tumor necrosis factor-α(all P<0.05);in the direct-contact co-culture system,HBV-specific CD8+T cells induced significant increases in the mortality rate of HepG2.2.15 cells(13.62%±3.27%vs 11.39%±2.40%,t=2.27,P=0.030)and the secretion of perforin and granzyme B(t=72.50,U=52.50,both P<0.05).In the direct-and indirect-contact co-culture systems,compared with HBV-specific CD8+T cells from the patients without HBsAg clearance,the HBV-specific CD8+T cells from patients with HBsAg clearance had a significantly greater reduction in HBV DNA(P<0.05)and significant increases in the secretion of interferon-γ and tumor necrosis factor-α(P<0.05).Conclusion PEG-IFN-α-2b therapy can help to achieve a relatively high HBsAg clearance rate in HBeAg-negative patients with chronic HBV infection,and the activity of HBV-specific CD8+T cells is significantly enhanced,which is closely associated with HBsAg clearance.关键词
聚乙二醇干扰素/乙型肝炎e抗原/乙型肝炎病毒/CD8阳性T淋巴细胞Key words
Pegylated Interferon/Hepatitis B e Antigens/Hepatitis B Virus/CD8-Positive T-Lymphocytes引用本文复制引用
秦蕾,李广鹏,申培君,张兰芳,杨晓飞,彭梅娟,张野..聚乙二醇干扰素-α-2b对HBeAg阴性慢性HBV感染者HBV特异性CD8+T淋巴细胞杀伤功能的影响[J].临床肝胆病杂志,2025,41(4):628-636,9.基金项目
空军军医大学科技发展基金(2022XB031)Science and Technology Development Funding of Air Force Military Medical University(2022XB031) （2022XB031）
