中华肝脏病杂志
中華肝髒病雜誌
중화간장병잡지
CHINESE JOURNAL OF HEPATOLOGY
2013年
8期
584-589
,共6页
韩亚萍%李军%蒋龙凤%徐庆庆%刘波%董莉%陈念%孔练花%谢发仁
韓亞萍%李軍%蔣龍鳳%徐慶慶%劉波%董莉%陳唸%孔練花%謝髮仁
한아평%리군%장룡봉%서경경%류파%동리%진념%공련화%사발인
肝炎,乙型,慢性%肝炎e抗原,乙型%白细胞介素-10%干扰素γ%细胞程序性死亡受体1
肝炎,乙型,慢性%肝炎e抗原,乙型%白細胞介素-10%榦擾素γ%細胞程序性死亡受體1
간염,을형,만성%간염e항원,을형%백세포개소-10%간우소γ%세포정서성사망수체1
Hepatitis B,chronic%Hepatitis B e Agtigens%Interleukin-10%Interferon-gamma%Programmed cell death 1
目的 探讨HBeAg对慢性乙型肝炎(CHB)患者外周血单个核细胞(PBMC)功能的调节作用. 方法 以重组的HBeAg体外刺激CHB患者和健康志愿者的PBMC,用流式细胞术和酶联免疫吸附试验法检测其刺激前后Th1/Th2型细胞因子的变化情况,并观察HBeAg对CHB患者PBMC表面细胞程序性死亡受体(PD)1及其配体(PD-L)1表达的影响.两组间资料比较采用独立样本t检验; PD-1/PD-L1表达水平与HBV DNA拷贝数的相关性采用Spearman相关分析.结果 HBeAg刺激后可使HBeAg阴性CHB患者和健康志愿者CD3+CD4+T淋巴细胞内干扰素(IFN)γ表达水平(0.17％±0.08％与0.17％±0.04％)明显低于未刺激组(0.30％±0.16％与0.32％±0.12％),t值分别为-2.382和-4.190,P值均＜0.01;培养上清液中白细胞介素(IL)-6、IL-10和肿瘤坏死因子α含量明显高于未刺激组(HBeAg阴性CHB患者的t值分别为2.504,3.583和4.324,健康志愿者t值分别为3.542,6.246和5.273,P值均＜0.01).HBeAg刺激PBMC后,HBeAg阴性CHB患者和健康志愿者CD14+细胞表面PD-L1表达水平分别为13.02％±4.98％和3.10％±2.47％,明显高于未刺激组的5.89％±1.56％和0.97％±0.83％,t值分别为4.815和3.454,P值均＜0.05.基础状态下在HBeAg阳性CHB患者外周血中,CD3+CD4+T淋巴细胞内IFNγ表达水平为0.23％±0.09％,明显低于HBeAg阴性CHB患者和健康志愿者的0.34％±0.15％和0.35％±0.09％(t=-3.177,P＜0.01 ; t=-4.541,P＜0.01);而IL-4表达水平为0.39％±0.16％,明显高于HBeAg阴性CHB患者和健康志愿者的0.26％±0.12％和0.23％±0.12％,t值分别为3.382和4.393,P值均＜0.01.基础状态下在HBeAg阳性CHB患者外周血中,CDB+T淋巴细胞表面PD-1和PD-L1表达水平明显高于HBeAg阴性CHB患者及健康志愿者(P值均＜ 0.01),CD14+T淋巴细胞表面PD-L1表达水平显著高于HBeAg阴性患者和健康志愿者,t值分别为5.092和5.473,P值均＜0.01 ; HBeAg阴性CHB患者外周血中CD3+T淋巴细胞表面PD-L1表达水平明显高于健康志愿者(t=3.214,P＜0.01).结论 HBeAg可以明显抑制Th1型细胞因子IFN γ的产生,促进Th2型细胞因子IL-6和IL-10分泌,上调外周血PBMC表面PD-1/PD-L1的表达,从而有利于形成对HBV感染的免疫耐受.因此,HBeAg可能是造成慢性HBV感染者体内免疫耐受的重要因素之一.
目的 探討HBeAg對慢性乙型肝炎(CHB)患者外週血單箇覈細胞(PBMC)功能的調節作用. 方法 以重組的HBeAg體外刺激CHB患者和健康誌願者的PBMC,用流式細胞術和酶聯免疫吸附試驗法檢測其刺激前後Th1/Th2型細胞因子的變化情況,併觀察HBeAg對CHB患者PBMC錶麵細胞程序性死亡受體(PD)1及其配體(PD-L)1錶達的影響.兩組間資料比較採用獨立樣本t檢驗; PD-1/PD-L1錶達水平與HBV DNA拷貝數的相關性採用Spearman相關分析.結果 HBeAg刺激後可使HBeAg陰性CHB患者和健康誌願者CD3+CD4+T淋巴細胞內榦擾素(IFN)γ錶達水平(0.17％±0.08％與0.17％±0.04％)明顯低于未刺激組(0.30％±0.16％與0.32％±0.12％),t值分彆為-2.382和-4.190,P值均＜0.01;培養上清液中白細胞介素(IL)-6、IL-10和腫瘤壞死因子α含量明顯高于未刺激組(HBeAg陰性CHB患者的t值分彆為2.504,3.583和4.324,健康誌願者t值分彆為3.542,6.246和5.273,P值均＜0.01).HBeAg刺激PBMC後,HBeAg陰性CHB患者和健康誌願者CD14+細胞錶麵PD-L1錶達水平分彆為13.02％±4.98％和3.10％±2.47％,明顯高于未刺激組的5.89％±1.56％和0.97％±0.83％,t值分彆為4.815和3.454,P值均＜0.05.基礎狀態下在HBeAg暘性CHB患者外週血中,CD3+CD4+T淋巴細胞內IFNγ錶達水平為0.23％±0.09％,明顯低于HBeAg陰性CHB患者和健康誌願者的0.34％±0.15％和0.35％±0.09％(t=-3.177,P＜0.01 ; t=-4.541,P＜0.01);而IL-4錶達水平為0.39％±0.16％,明顯高于HBeAg陰性CHB患者和健康誌願者的0.26％±0.12％和0.23％±0.12％,t值分彆為3.382和4.393,P值均＜0.01.基礎狀態下在HBeAg暘性CHB患者外週血中,CDB+T淋巴細胞錶麵PD-1和PD-L1錶達水平明顯高于HBeAg陰性CHB患者及健康誌願者(P值均＜ 0.01),CD14+T淋巴細胞錶麵PD-L1錶達水平顯著高于HBeAg陰性患者和健康誌願者,t值分彆為5.092和5.473,P值均＜0.01 ; HBeAg陰性CHB患者外週血中CD3+T淋巴細胞錶麵PD-L1錶達水平明顯高于健康誌願者(t=3.214,P＜0.01).結論 HBeAg可以明顯抑製Th1型細胞因子IFN γ的產生,促進Th2型細胞因子IL-6和IL-10分泌,上調外週血PBMC錶麵PD-1/PD-L1的錶達,從而有利于形成對HBV感染的免疫耐受.因此,HBeAg可能是造成慢性HBV感染者體內免疫耐受的重要因素之一.
목적 탐토HBeAg대만성을형간염(CHB)환자외주혈단개핵세포(PBMC)공능적조절작용. 방법 이중조적HBeAg체외자격CHB환자화건강지원자적PBMC,용류식세포술화매련면역흡부시험법검측기자격전후Th1/Th2형세포인자적변화정황,병관찰HBeAg대CHB환자PBMC표면세포정서성사망수체(PD)1급기배체(PD-L)1표체적영향.량조간자료비교채용독립양본t검험; PD-1/PD-L1표체수평여HBV DNA고패수적상관성채용Spearman상관분석.결과 HBeAg자격후가사HBeAg음성CHB환자화건강지원자CD3+CD4+T림파세포내간우소(IFN)γ표체수평(0.17％±0.08％여0.17％±0.04％)명현저우미자격조(0.30％±0.16％여0.32％±0.12％),t치분별위-2.382화-4.190,P치균＜0.01;배양상청액중백세포개소(IL)-6、IL-10화종류배사인자α함량명현고우미자격조(HBeAg음성CHB환자적t치분별위2.504,3.583화4.324,건강지원자t치분별위3.542,6.246화5.273,P치균＜0.01).HBeAg자격PBMC후,HBeAg음성CHB환자화건강지원자CD14+세포표면PD-L1표체수평분별위13.02％±4.98％화3.10％±2.47％,명현고우미자격조적5.89％±1.56％화0.97％±0.83％,t치분별위4.815화3.454,P치균＜0.05.기출상태하재HBeAg양성CHB환자외주혈중,CD3+CD4+T림파세포내IFNγ표체수평위0.23％±0.09％,명현저우HBeAg음성CHB환자화건강지원자적0.34％±0.15％화0.35％±0.09％(t=-3.177,P＜0.01 ; t=-4.541,P＜0.01);이IL-4표체수평위0.39％±0.16％,명현고우HBeAg음성CHB환자화건강지원자적0.26％±0.12％화0.23％±0.12％,t치분별위3.382화4.393,P치균＜0.01.기출상태하재HBeAg양성CHB환자외주혈중,CDB+T림파세포표면PD-1화PD-L1표체수평명현고우HBeAg음성CHB환자급건강지원자(P치균＜ 0.01),CD14+T림파세포표면PD-L1표체수평현저고우HBeAg음성환자화건강지원자,t치분별위5.092화5.473,P치균＜0.01 ; HBeAg음성CHB환자외주혈중CD3+T림파세포표면PD-L1표체수평명현고우건강지원자(t=3.214,P＜0.01).결론 HBeAg가이명현억제Th1형세포인자IFN γ적산생,촉진Th2형세포인자IL-6화IL-10분비,상조외주혈PBMC표면PD-1/PD-L1적표체,종이유리우형성대HBV감염적면역내수.인차,HBeAg가능시조성만성HBV감염자체내면역내수적중요인소지일.
Objective To study the immunoregulatory effect of hepatitis B virus (HBV) e antigen (HBeAg) on peripheral blood monocytes (PBMCs).Methods PBMCs were isolated from patients with chronic hepatitis B (CHB; both HBeAg-and HBeAg+) and healthy controls,and cultured with recombinant HBeAg.The HBeAg-induced changes in expression of PD-1/PD-L1 were measured by flow cytometry of the cells and in secreted cytokines were measured by enzyme-linked immunosorbent assay of the supernatants.Comparisons between two groups were made by the independent-samples t-test;the relationship between PD-1/B7-H1 level and HBV DNA copy number was evaluated by Spcarman's correlation analysis.Results Exposure to HBeAg led to a significant decrease in CD3+CD4+ T lymphocyte-specific expression of IFNα for both the CHB patients' and healthy controls' samples (t =2.382 and-4.190 respectively,P ＜ 0.01).For the HBeAg-CHB patients' and healthy controls' samples,the HBeAg exposure led to increased levels of secreted cytokines IL-6,IL-10 and TNFα (t =2.504,3.583 and 4.324,P ＜ 0.01 and t =3.542,6.246 and 5.273,P ＜ 0.01 respectively) and of CD14+ PBMC-specific expression of PD-L1 (t =4.815 and 3.454,P ＜ 0.05 respectively).Compared to the HBeAg-negative CHB patients' and healthy controls' samples,the HBeAg+ CHB patients' samples had significantly lower CD3+CD4+ T cell-specific expression of IFNα (t =-3.177 and-4.541,P ＜ 0.01 respectively),but significantly higher levels of secreted IL-4 (t =3.382 and 4.393,P ＜ 0.01 respectively),ofCD3+ T cells-specific expression of PD-1/PD-L1 (t =4.755,2.942 and 4.518,4.595,P ＜ 0.01 respectively),and of CD14+ T cells-specific expression of PD-L1 (t =5.092 and 5.473,P ＜ 0.01 respectively).The CD3+ T cells-specific expression of PD-L1 was significantly higher in the samples from HBeAg-CHB patients than from the healthy controls (t =3.214,P ＜ 0.01).Conclusion HBeAg was able to down-regulate the production ofTh1-type eytokines (IFNγ),and up-regulate the secretion of Th2-type cytokines (IL-6,IL-10) and the expression of PD-1/PD-L1on monocytes.These changes are conducive to the formation of immune tolerance to HBV.Therefore,HBeAg may play an important role in immune tolerance to chronic HBV infection.
