HBcAg特异性Th9细胞在乙型肝炎病毒感染者中的免疫调控作用  

HBcAg-specific Th9 cells in patients with HBV infection

作　　者：段树鹏[1] 朱利红[2] 侯丽娟[1] 王宏伟[1] 宋新文[1] 郝洁[3] 孙淑卫 申保生[1] Duan Shupeng;Zhu Lihong;Hou Lijuan;Wang Hongwei;Song Xinwen;Hao Jie;Sun Shuwei;Shen Baosheng(Department of Infectious Diseases,the First Affiliated Hospital of Xinxiang Medical University,Xinxiang 453100,China;Department of Obstetrics and Gynecology,the First Affiliated Hospital of Xinxiang Medical University,Xinxiang 453100,China;Medical Administration Division,the First Affiliated Hospital of Xinxiang Medical University,Xinxiang 453100,China)

机构地区：[1]新乡医学院第一附属医院感染科,453100 [2]新乡医学院第一附属医院妇产科,453100 [3]新乡医学院第一附属医院医务科,453100

出　　处：《中华微生物学和免疫学杂志》2021年第8期608-615,共8页Chinese Journal of Microbiology and Immunology

基　　金：河南省医学科技攻关计划项目(201303106)。

摘　　要：目的观察乙型肝炎病毒(hepatitis B virus,HBV)感染者中非特异性和乙型肝炎核心抗原(HBcAg)特异性Th9细胞、白细胞介素-9(IL-9)的变化及其对CD8+T细胞功能的影响。方法纳入2018年1月至2019年1月在新乡医学院第一附属医院就诊的急性乙型肝炎(acute hepatitis B,AHB)患者12例、慢性乙型肝炎(chronic hepatitis B,CHB)患者58例、健康对照者20例,分离外周血单个核细胞(peripheral blood mononuclear cells,PBMCs)和血浆,流式细胞术检测非特异性Th9细胞(CD3+CD4+IL-9+)和HBcAg特异性Th9细胞,ELISA检测血浆IL-9水平。CHB患者接受富马酸替诺福韦二吡呋酯片(tenofovir disoproxil fumarate,TDF)抗病毒治疗,观察抗病毒治疗48周时非特异性Th9细胞、HBcAg特异性Th9细胞和血浆IL-9的变化。分选12例HLA-A2限制性CHB患者的CD4^(+)CCR4-CCR6-CXCR3-细胞(即Th9细胞)和CD8^(+)T细胞,与HepG2.2.15细胞共培养,同时在培养液中加入抗IL-9中和抗体,检测HepG2.2.15细胞死亡比例和IFN-γ、TNF-α水平。组间比较采用t检验、单因素方差分析或SNK-q检验。结果非特异性Th9细胞和血浆IL-9水平在AHB患者、CHB患者和对照者之间的差异无统计学意义(P>0.05),CHB患者中HBcAg特异性Th9细胞低于AHB患者[(2.49±0.61)%vs(3.19±0.62)%,P<0.001],CHB患者中HBcAg特异性Th9细胞与HBV DNA呈显著负相关(r=-0.385,P=0.003),但与丙氨酸转氨酶(ALT)无显著相关性(P>0.05)。TDF抗病毒治疗48周可显著升高CHB患者HBcAg特异性Th9细胞比例[(2.94±0.48)%,P<0.001],但对非特异性Th9细胞和血浆IL-9水平无显著影响(P>0.05)。CHB患者HBcAg特异性Th9细胞的杀伤活性低,但可诱导CD8^(+)T细胞对HepG2.2.15细胞杀伤作用增强,主要表现为HepG2.2.15细胞死亡比例增加、IFN-γ和TNF-α水平升高,抗IL-9中和抗体可降低HBcAg特异性Th9细胞对CD8+T细胞杀伤功能的增强作用(P<0.001)。结论慢性HBV感染可能抑制HBcAg特异性Th9细胞水平和功能,诱导感染慢性化。Objective To investigate the changes of non-specific and HBV core antigen(HBcAg)-specific Th9 cells,and intereleukin-9(IL-9)in HBV-infected patients,and to assess the influence of Th9 cells on CD8+T cell function.Methods Twelve patients with acute hepatitis B(AHB)and 58 with chronic hepatitis B(CHB),who were hospitalized in the First Affiliated Hospital of Xinxiang Medical University between January 2018 and January 2019,were enrolled in this study.Twenty healthy subjects negative for HBsAg were selected as controls.Peripheral blood mononuclear cells(PBMCs)and plasma samples were isolated.Non-specific Th9 cells(CD3^(+)CD4^(+)IL-9^(+))and HBcAg-specific Th9 cells were analyzed by flow cytometry.Plasma IL-9 level was measured by enzyme linked immunosorbent assay.CHB patients received tenofovir disoproxil fumarate(TDF)antiviral therapy.The changes of non-specific Th9 cells,HBcAg-specific Th9 cells and plasma IL-9 level were assessed 48 weeks after TDF therapy.CD4^(+)CCR4-CCR6-CXCR3-(Th9)cells and CD8^(+)T cells were isolated from 12 HLA-A2 restricted CHB patients and co-cultured with HepG2.2.15 cells with the presence of anti-IL-9 neutralizing antibody.The percentage of dead HepG2.2.15 cells and the levels of IFN-γand TNF-αwere detected.Student′s t test,one-way analysis of variance or SNK-q test was used for statistical comparison between groups.Results There were no significant differences in non-specific Th9 cells or plasma IL-9 level among AHB patients,CHB patients and healthy controls(P>0.05).HBcAg-specific Th9 cells was down-regulated in CHB patients when compared with AHB patients[(2.49±0.61)%vs(3.19±0.62)%,P<0.001].The percentage of HBcAg-specific Th9 cells was negatively correlated with HBV DNA(r=-0.385,P=0.003),but not correlated with ALT(P>0.05)in CHB patients.TDF therapy for 48 weeks remarkably elevated the HBcAg-specific Th9 cells[(2.94±0.48)%,P<0.001],however,did not affect non-specific Th9 cells or plasma IL-9 level(P>0.05)in CHB patients.The cytotoxicity of HBcAg-specific Th9 cells was low in C

关 键 词：乙型肝炎 T淋巴细胞 免疫应答 

分 类 号：R512.62[医药卫生—内科学]