慢性乙型肝炎病毒感染者外周血单个核细胞表达程序性死亡分子-1及其配体与血清乙型肝炎病毒DNA水平的相关性

Objective To study the relationship between programmed death-1 (PD-1)/programmed death-1 ligand (PD-L1) expressions and serum hepatitis B virus (HBV) DNA levels in chronic hepatitis B (CHB) patients. Methods A total of 137 CHB patients and 10 healthy controls were enrolled in the study. The peripheral blood mononuclear cells (PBMCs) were isolated from fresh blood samples. HBV-specific cytotoxic T lymphocyte (CTL) was expanded in vitro in 64 human leucocyte antigen (HLA)-A2 positive patients. Flow cytometry was used to detect HLA-A2 type,expressions of PD-1/PD-L1 on PBMCs and PD-1 on HBV specific CTL. Interferon gamma (IFN-γ)was measured by commercial enzyme-linked immunosorbent assay (ELISA) kits. PD-1/PD-L1expressions on PBMCs, HBV-specific CTL and IFN-γ level in PBMC culture medium were compared among patients with different baseline HBV DNA levels. Ten hepatitis B e antigen (HBeAg) positive patients were treated with telbivudine for 24 weeks. The above mentioned parameters were determined and compared before and after the antiviral treatment. Independent-samples t test were used to compare means between two groups and one-way A NOVA were used to compare means among multigroups. We used the pearson corretation test to assess corretation significance. Results The PD-1 and PD-L1 expressions on PBMCs in patients with baseline HBV DNA＜3 lg copy/mL, 3-6 lg copy/mL and ＞6 lg copy/mL were all significant higher than those in healthy control group, but no statistical differences were found. PD-1 expressions on HBV-specific CTL in the three CHB patient groups were (69.3±11.2)%, (76.5±9. 1)% and (78.0±11.7)%, respectively. However, PD-1 expression on HBV-specific CTL was higher, while the frequency of HBV-specific CTL cells was lower in HBV DNA ＞6 lg copy/mL group compared to HBV DNA＜3 lg copy/mL group. The above parameters, including expressions of PD-1 and PD-L1, the frequency of HBV-specific CTL and its PD-1 expression were not significantly different between HBeAg-positive group and HBeAg-negative group. Compared with baseline, PD-1 and PD-L1 expression decreased obviously accompanying with increase of HBV-specific CTL cells frequency and IFN-γ level after 12 weeks and 24 weeks of telbivudine treatment. Conclusions PD-1 expression on HBV-specific CTL correlates with serum HBV DNA level, but not HBeAg status in CHB patients. Suppression of HBV replication can reduce PD-1/PD-L1 expressions and partially restore HBV specific CTL function.     

慢性乙型肝炎病毒感染者外周血单个核细胞表达程序性死亡分子-1及其配体与血清乙型肝炎病毒DNA水平的相关性

Objective To study the relationship between programmed death-1 (PD-1)/programmed death-1 ligand (PD-L1) expressions and serum hepatitis B virus (HBV) DNA levels in chronic hepatitis B (CHB) patients. Methods A total of 137 CHB patients and 10 healthy controls were enrolled in the study. The peripheral blood mononuclear cells (PBMCs) were isolated from fresh blood samples. HBV-specific cytotoxic T lymphocyte (CTL) was expanded in vitro in 64 human leucocyte antigen (HLA)-A2 positive patients. Flow cytometry was used to detect HLA-A2 type,expressions of PD-1/PD-L1 on PBMCs and PD-1 on HBV specific CTL. Interferon gamma (IFN-γ)was measured by commercial enzyme-linked immunosorbent assay (ELISA) kits. PD-1/PD-L1expressions on PBMCs, HBV-specific CTL and IFN-γ level in PBMC culture medium were compared among patients with different baseline HBV DNA levels. Ten hepatitis B e antigen (HBeAg) positive patients were treated with telbivudine for 24 weeks. The above mentioned parameters were determined and compared before and after the antiviral treatment. Independent-samples t test were used to compare means between two groups and one-way A NOVA were used to compare means among multigroups. We used the pearson corretation test to assess corretation significance. Results The PD-1 and PD-L1 expressions on PBMCs in patients with baseline HBV DNA＜3 lg copy/mL, 3-6 lg copy/mL and ＞6 lg copy/mL were all significant higher than those in healthy control group, but no statistical differences were found. PD-1 expressions on HBV-specific CTL in the three CHB patient groups were (69.3±11.2)%, (76.5±9. 1)% and (78.0±11.7)%, respectively. However, PD-1 expression on HBV-specific CTL was higher, while the frequency of HBV-specific CTL cells was lower in HBV DNA ＞6 lg copy/mL group compared to HBV DNA＜3 lg copy/mL group. The above parameters, including expressions of PD-1 and PD-L1, the frequency of HBV-specific CTL and its PD-1 expression were not significantly different between HBeAg-positive group and HBeAg-negative group. Compared with baseline, PD-1 and PD-L1 expression decreased obviously accompanying with increase of HBV-specific CTL cells frequency and IFN-γ level after 12 weeks and 24 weeks of telbivudine treatment. Conclusions PD-1 expression on HBV-specific CTL correlates with serum HBV DNA level, but not HBeAg status in CHB patients. Suppression of HBV replication can reduce PD-1/PD-L1 expressions and partially restore HBV specific CTL function.     

慢性乙型肝炎患者外周血CD8+T淋巴细胞表面程序性死亡受体-1表达上调的意义

程序性死亡受体-1(PD-1)主要表达于活化的淋巴细胞和单核细胞,尤其是体内活化的T淋巴细胞表面[1],负性调节其活化、增殖和细胞因子的产生[2-3],使人类免疫缺陷病毒(HIV)[4]、HCC[5]、HBV[6-7]等慢性感染过程中患者的病毒特异性CD8+细胞毒性T淋巴细胞(CTL)功能受到抑制,从而造成持续性感染状态.     

慢性乙型肝炎抗病毒疗效与外周血单个核细胞内乙型肝炎病毒DNA水平的关系

目的 探讨慢性乙型肝炎(CHB)抗病毒疗效与达到停药标准时外周血单个核细胞(PBMC)内HBV DNA水平的关系.方法 入选90例经抗病毒治疗达到停药标准的CHB患者,其中应用IFN 44例,应用核苷类药物46例.所有患者均于停药时检测PBMC内HBV DNA,比较阴性组和阳性组治疗前血清HBV DNA水平与达到停药标准时PBMC内HBV DNA的关系,观察停药时PBMC内HBV DNA水平与复发的关系.计量资料采用t检验,计数资料采用X2检验.结果 90例CHB患者停药时,PBMC内HBV DNA阴性组67例,阳性组23例.CHB患者血清HBV DNA阳转率在PBMC内HBV DNA阴性组为13.4%(9/67例),显著低于阳性组的73.9%(17/23例),差异有统计学意义(X2=30.4873,P<0.01).PBMC内HBV DNA阴性组与阳性组在肝病复发ALT升高幅度(t=0.8729,P=0.3913)、停药后复发时间(t=1.9222,P=0.0665)均差异无统计学意义,而在血清HBV DNA反弹幅度则差异有统计学意义(t=2.7493,P=0.0112).5例患者获得HBsAg血清学转换,且均未检测到PBMC内HBV DNA,随访6～12个月无一例复发.PBMC内HBV DNA阳性组治疗前血清HBV DNA水平为(7.2±1.1)lg拷贝/mL,显著高于阴性组的(5.2±2.1)lg拷贝/mL(t=4.3557,P<0.01).结论 经抗病毒治疗达到停药标准的CHB患者,其停药时的PBMC内HBV DNA水平可能是预测抗病毒疗效持久性的重要因素之一.

Abstract：

Objective To explore the relationship between the antiviral effect and peripheral blood mononuclear cell (PBMC) hepatitis B virus (HBV) DNA when the patients reach the standard of withdrawal of antiviral therapy in chronic hepatitis B (CHB).Methods Ninety CHB patients treated with interferon(n=44) or nucleot (s) ide(n=46) who reached the standard of withdrawal of antiviral therapy were recruited.HBV DNA levels in PBMCs were tested at the end of treatment,and its relationship with serum HBV DNA level before treatment in PBMC HBV DNA positive group and negative group were compared.The correlation between HBV DNA in PBMCs at the end of treatment and relapse were explored.Measurement data were analyzed by student t test and enumeration data were analyzed by X2 test.Results Among 90 patients,67(74.4%) were PBMC HBV DNA negative at the end of treatment,and 23(25.6%) were positive.The serum HBV DNA positive conversion rate in PBMC HBV DNA negative patients was 13.4%,which were significantly lower than that in positive group (73.9%) (X2=30. 4873, P<0.01 ). There were no significant differences of alanine aminotransferase (ALT) levels when hepatitis flare (t=0. 8729, P=0. 3913) and relapse time (t=1. 9222, P=0. 0665) between PBMC HBV DNA negative group and positive group after withdrawal of therapy, while the serum HBV DNA rebound was greater in positive group than that in negative group (t=2. 7493, P=0. 0112). There were five patients who achieved hepatitis B surface antigen (HBsAg) seroconversion, whose PBMC HBV DNA were all undetectable, and none relapsed during follow-up for 6-12 months. The pretreatment HBV DNA as level in PBMC HBV DNA positive was (7.2±1.1) lg copy/mL, which was much higher than that in negative group[(5.2±2.1) lg copy/mL] (t=4. 3557, P<0.01). Conclusions In patients who reach the standard of drug withdrawal,PBMC HBV DNA at the end of treatment is an important predictor for durability of antiviral therapy in CHB.     

芪灵合剂对慢性乙型肝炎患者T细胞表面程序性死亡1和程序性死亡配体1表达的影响

目的：观察中药芪灵合剂对慢性乙型肝炎（CHB）患者外周血CD4＋T淋巴细胞表面程序性死亡1（Programmed death-1,PD-1）及程序性死亡配体1（Programmed death-1 ligand1,PD-L1）表达水平的影响。方法：收集56例CHB患者随机分为芪灵合剂联合拉米夫定片（LAM）治疗组29例和单用LAM对照组27例,于抗病毒治疗的第0、24、48周用流式细胞术检测外周血CD4＋T淋巴细胞及其表面的PD-1/PD-L1的表达情况。结果：治疗48周后,治疗组患者HBV DNA转阴率、ALT复常率均高于对照组,与对照组比较,HBV DNA转阴率具有统计学意义（P〈0.05）。治疗组患者外周血CD4＋T细胞表面PD-1 24周、48周表达水平与治疗前比较逐渐下降;48周时与本组治疗前比较差异有统计学意义（P〈0.01）,与对照组比较差异有统计学意义（P〈0.05）。治疗组患者CD4＋T细胞PD-L1 48周后表达水平与治疗前比较差异有统计学意义（P〈0.01）,与对照组比较差异有统计学意义（P〈0.05）。结论：中药芪灵合剂联合LAM能够提高CHB患者HBV DNA转阴率,ALT复常率,其作用机制可能与抑制CD4＋T淋巴细胞表面PD-1/PD-L1的表达有关。     

慢性乙型肝炎患者外周血单核细胞亚群表面程序性死亡配体1表达

目的 探讨程序性死亡配体1(PD-L1)在慢性乙型肝炎患者外周血单核细胞亚群表面的表达情况.方法 利用荧光抗体标记结合多色流式技术,检测慢性乙型肝炎患者外周血单核细胞各亚群表面PD-L1分子的表达.结果 慢性乙型肝炎患者外周血单核细胞PD-L1的表达率与血清AST水平呈正相关(r=0.358,P=0.035),且免疫活...     

慢性乙型肝炎病毒感染者程序性死亡因子1基因拷贝数和mRNA的表达水平

目的 研究慢性HBV感染者程序性死亡因子1(PD-1)基因的拷贝数和mRNA表达水平,以及在不同临床表现患者间的差异。 方法 实时荧光定量PCR法检测健康对照者27例（对照组）、慢性HBV携带者31例（携带组）、慢性重型肝炎患者19例（重肝组）、乙型肝炎相关性原发性肝癌患者29例（肝癌组）的外周血单个核细胞PD-1基因...     

抗病毒治疗慢性丙型肝炎患者T细胞表面程序性死亡1和程序性死亡配体1的表达

目的 观察慢性丙型肝炎(CHC)患者抗病毒治疗24周时外周血CD4+和CD8+T淋巴细胞(T细胞)表面表面程序性死亡1 (PD-1)和程序性死亡配体1(PD-L1)表达水平,分析其与抗病毒治疗临床转归的关系.方法 24例CHC患者,均采用聚乙二醇干扰素α-2a (Peg-IFN α-2a)每周皮下注射一次,联合利巴韦林800 ～ 1200 mg/d,治疗24 ～ 48周.采用流式细胞术和实时荧光定量检测患者治疗前、治疗4、12、24周外周血CD4+和CD8+T细胞表面PD-1、PD-L1表达水平和外周血HCV RNA,全自动生化分析仪检测ALT.采用SPSS16.0软件.两样本计量结果分析采用t检验,治疗前后的计量结果采用重复测量的单因素或两因素方差分析,所有检验为双侧检验. 结果 CHC患者治疗后4周HCV RNA阴性者19例,CD4+和CD8+T细胞表面PD-1的表达率在治疗前分别为18.6％±6.1％和16.6％±13.8％,治疗24周时分别为10.3％±7.7％和9.4％±4.6％,治疗前后比较,PD-1的表达明显下降,F值为12.406和4.955,P值为0.002和0.039,差异有统计学意义.CD8+T细胞表面PD-L1的表达率在治疗前为17.5％±13.7％,治疗4、12、24周时分别为25.9％±11.1％、29.6％±15.1％、32.0％±15.7％,治疗后明显升高,F值分别为9.063、8.365、9.736,P值均＜0.01.治疗4周时,HCV RNA阳性者5例,仅发现CD8+T细胞表面PD-L1的表达治疗24周(39.2％±15.6％)与治疗前(17.4％±16.7％)比较明显升高,F=10.292,P=0.033.持续病毒学应答者20例:CD4+T细胞表面PD-1的表达在治疗4、12、24周分别为14.4％±7.5％、14.0％±6.9％、10.7％±7.6％,治疗前为20.2％±7.5％,与治疗前比较明显下降,F值分别为6.133、5.541、14.780,P＜0.05或P＜0.01.CD8+T细胞表面PD-1的表达在治疗12、24周分别为10.2％±4.6％和10.1％±4.9％,治疗前为16.8％±13.4％,治疗前后比较,PD-1的表达在治疗后明显下降,F值为4.964和4.613,P值均＜0.05.CD8+T细胞表面PD-L1的表达在治疗12、24周分别为30.8％±16.6％和35.2％±16.5％,治疗前为19.0％±14.5％,治疗后明显升高,F=6.442,P=0.020和F=12.349,P=0.002.复发组4例,各治疗时间点PD-1和PD-L1与治疗前比较,差异无统计学意义.结论 快速有效的抗病毒治疗可以下调CHC患者外周血CD4+和CD8+T细胞表面PD-1的表达,上调CD8+T细胞表面PD-L1的表达.CHC患者外周血CD4+和CD8+T细胞表面PD-1和PD-L1表达水平的变化可能与患者抗病毒治疗临床转归存在关系.     

慢性乙型肝炎患者树突状细胞诱导的HBV特异性细胞毒性T淋巴细胞PD-1的表达

目的 探讨慢性乙型肝炎(CHB)患者树突状细胞(DC)诱导的HBV特异性细胞毒性T细胞(CTL)表面程序性死亡受体1(PD-1)的表达情况及其与HBV DNA的关系.方法 采集30例CHB患者和10例健康人的抗凝外周静脉血,分离外周血单个核细胞(PBMC),在白细胞介素(IL)-4和粒-巨噬细胞集落刺激因子(GM-CSF)的作用下培养使DC增殖、成熟,培养第4d加入纯化的HBsAg进行冲击.采同一患者外周血,分离出自体T淋巴细胞,用含重组人白细胞介素(rhIL)-2的培养基维持T细胞的生长,培养第5d与HBsAg冲击的DC共培养.以流式细胞技术检测CTL的PD-1表达,并分析PD-1表达水平与HBV DNA的关系.结果 与健康对照组比较,CHB组DC诱导的HBV特异CTL的PD-1的表达明显升高(P=0.000).且HBeAg阳性组PD-1的表达率较HBeAg阴性组明显升高(P=0.000).CHB患者DC诱导的HBV特异性CTL的PD-1表达率与血清HBV DNA拷贝数的对数值呈正相关(r=0.53,P=0.008).结论 CHB患者DC诱导的HBV特异性CTL高表达PD-1分子,为HBV慢性感染过程中CTL功能低下,病毒难以清除提供了另一条重要线索.     

急性乙型肝炎患者程序性细胞死亡受体1表达与记忆性T淋巴细胞形成的关系

目的 观察急性自限性乙型肝炎发病过程中患者体内病毒抗原特异性细胞毒性T淋巴细胞(CTL)上程序性细胞死亡受体1(PD-1)表达的动态变化特点及其与记忆性抗原特异性CD8+T淋巴细胞形成的关系. 方法 针对不同表位合成4种五聚体,长期随访收集11例人类白细胞抗原(HLA)-A2阳性的急性乙型肝炎患者的外周血,流式细胞仪检测病毒特异性CTL上免疫抑制性分子PD-1、记忆性分子(CCR7、CD45RA、CD127)和活化标志物CD38的表达情况,并分析其相关性.同时进行肝功能,HBsAg、抗-HBs和血清HBV DNA载量检测.结果所有急性自限性乙型肝炎患者发病早期均表现出高频度、病毒抗原多表位的特异性CTL反应,而晚期各表位CTL频率均明显下降.CTL上PD-1表达在早期明显上调;与早期比较,晚期PD-1分子的表达明显降低(t=4.314,P＜0.01).同时CTL高表达记忆性分子CCR7,CD45RA和CD127,而低表达活化标志物CD38.提示病毒清除后记忆性CD8+T淋巴细胞形成. 结论 在急性自限性乙型肝炎发病过程中,HBV特异性CTL上PD-1分子表达的动态变化与记忆性T淋巴细胞的形成密切相关.     

B7-H1及其受体PD-1在慢性乙型肝炎患者淋巴细胞上的表达及意义

目的研究B7-H1及其受体PD-1在慢性乙型肝炎患者T淋巴细胞及髓样树突细胞（mDCs）上的表达及它们的表达水平与患者疾病状态的关系。方法流式细胞技术检测正常人和慢性乙型肝炎患者CD4^＋、CD8^＋T淋巴细胞及mDCs上B7-H1和PD-1的表达水平。实时定量PCR检测患者的HBV DNA。结果慢性乙型肝炎患者B7-H1及其受体PD-1的表达水平明显升高,健康对照mDCs、CD4^＋及CD8^＋T淋巴细胞B7-H1的阳性表达率分别为0.35%±0.10%、3.63%±0.70%和1.20%±0.19%,慢性乙型肝炎患者分别为7.88%±1.40%、24.28%±2.86%和10.78%±1.62%,慢性乙型肝炎患者B7-H1在mDCs和T淋巴细胞上的表达水平明显高于健康对照（P值均〈0.05）健康对照CD4^＋及CD8^＋T淋巴细胞PD-1的阳性表达率分别为5.92%±1.75%和5.98%±0.88%,慢性乙型肝炎患者分别为17.76%±2.47%和11.92%±2.21%,慢性乙型肝炎患者PD-1在T淋巴细胞上的表达水平也明显高于健康对照（P值均〈0.05）。且它们的表达与患者的ALT水平及HBV DNA载量呈明显的正相关（P值均〈0.05）。结论慢性乙型肝炎患者淋巴细胞上B7-H1和PD-1的表达水平与患者疾病状态密切相关。     

乙型肝炎病毒总DNA、共价闭合环状DNA和HBsAg在各种慢性乙型肝炎病毒感染者中的定量检测

目的 定量检测慢性乙型肝炎(CHB)、乙型肝炎肝硬化(LC)、肝细胞癌(HCC)患者HBV总DNA(tDNA)、HBV共价闭合环状DNA (cccDNA)和HBsAg,并探讨其特点.方法 荧光定量PCR检测21例CHB、23例LC和25例HCC患者外周血和肝组织标本HBV tDNA、HBVcccDNA,化学发光法定量检测外周血HBsAg.正态数据采用ANOVA分析和t检验,相关性分析采用Pearson检验,非正态数据采用秩和检验.结果 在CHB、LC和HCC患者中,外周血HBVtDNA分别为(5.38±2.08)、(4.96±1.65)和(4.18±0.91)lg拷贝/mL,肝组织HBV tDNA分别为(7.18±1.91)、(6.51±1.87)和(5.87±1.47)lg拷贝/μg,肝组织HBV cccDNA分别为(3.53±2.03)、(2.63±2.13)和(0.58±1.40)lg拷贝/μg,外周血H BsAg分别为(3.30±0.65)、(3.12±0.52)和(2.60±1.03)lg IU/mL,CHB与HCC患者比较,差异均有统计学意义(t=2.446,P=0.013;t=2.562,P=0.014;t=5.799,P＜0.01;t=2.709,P=0.003),LC与HCC患者肝组织HBVcccDNA及HBsAg定量比较,差异有统计学意义(t=-3.894,P＜0.01;t=-2.237,P=0.023).外周血均未检出HBV cccDNA.HBsAg定量与外周血HBV tDNA(r=-0.290,P=0.016)、肝组织HBV tDNA(r=0.372,P=0.002)及肝组织HBV cccDNA(r=0.378,P=0.001)均有关.结论 HBV tDNA、HBV cccDNA、HBsAg在CHB、LC、HCC患者呈逐渐降低趋势,HBsAg定量与外周血HBV tDNA、肝组织HBV tDNA及肝组织HBV cccDNA有关.     

"免疫耐受期"慢性乙型肝炎病毒感染者临床与肝脏病理学特点

目的 了解免疫耐受期慢性HBV感染者临床及肝脏病理学特征.方法 分析98例"免疫耐受期"慢性HBV患者年龄、性别、血清HBV DAN水平、肝脏炎症活动度及纤维化程度、肝脏HBsAg和HBcAg表达情况.并对不同血清ALT水平的肝脏炎症活动度及纤维化程度进行比较.采用X2检验.结果 98例患者中,＜30岁者83例,占84.7%,≥30岁者15例,占15.3%.有48.0%感染者的母亲HBsAg阳性.所有感染者血清HBVDNA为高水平复制,＞1×107拷贝/mL者占78.5%.仅5例感染者肝脏炎症活动度为Go,占5.1%;其中G1 64例,占65.3%;G2 29例,占29.6%.56例肝脏无明显纤维化(SO),占57.1%;S1 23例,占23.5%;S2 14例,占14.3%;S3 5例,占5.1%;未发现肝硬化.肝组织HBsAg阳性79例,占80.6%;HBeAg阳性80例,占81.6%.血清ALT在正常范围高水平的感染者肝脏纤维化程度明显高于ALT低水平者(X2=8.112 3,P=0.043 7).结论绝大部分"免疫耐受期"慢性HBV感染者肝脏存在轻度炎性反应,部分已出现纤维化;对"免疫耐受期"HBV感染者进行肝脏病理学检查,有利于正确判断病情和确定治疗方案.     

T淋巴细胞亚群及HBV特异性细胞毒性T淋巴细胞的变化在慢性乙型肝炎重症化中的意义

我们观察了209例慢性乙型肝炎(CHB)住院患者进展为慢性重型乙型肝炎前、后的HBV特异性细胞毒性T淋巴细胞(CTL)、非特异性CTL、T淋巴细胞(简称T细胞)亚群的变化,为预防或阻止CHB患者进展为重型肝炎提供依据.     

HBeAg状态及乙型肝炎病毒载量对慢性重型乙型肝炎预后的影响

目的 探讨HBeAg状态及HBV DNA载量对慢性重型乙型肝炎预后的影响.方法 回顾分析2002年1月至2007年12月在南方医科大学南方医院住院的慢性重型乙型肝炎患者406例,研究HBeAg状态、HBV DNA载量对疾病预后的影响.计量资料采用t检验,率的比较采用X2检验.结果 406例重型肝炎患者中,HBeAg阳性208例,占51.2%,HBeAg阴性198例,占48.8%.HBeAg阳性组与HBeAg阴性组比较,两组间男女构成比、TBil峰值及平均凝血酶原活动度谷值差异均无统计学意义;HBeAg阴性组平均年龄(46.7±12.8)岁,显著高于HBeAg阳性组(38.3±13.5)岁(t=6.43,P＜0.01);HBeAg阴性组肝硬化患者占67.7%,亦显著高于HBeAg阳性组的45.7%(X2=19.97,P＜0.01);HBeAg阴性组好转率为32.3%,显著低于HBeAg阳性组的44.7%(X2=6.56,P＜0.05).在208例HBeAg阳性与198例HBeAg阴性患者中,均显示随着HBV DNA载量的升高,其好转率下降,呈显著负相关(X2=22.98,X2=26.04;均P＜0.01).结论 HBeAg阴性重型乙型肝炎较HBeAg阳性者预后差;无论HBeAg状态如何,HBV DNA载量越高,其预后越差.