乙型肝炎患者外周血病毒特异性CD8阳性细胞毒性T淋巴细胞的数量和功能分析

目的 分析急性乙型肝炎(AHB)和慢性乙型肝炎(CHB)患者外周血中乙型肝炎病毒(HBV)特异性细胞毒性T淋巴细胞( CTL)的数量及功能.方法 36例HLA-A2阳性的乙型肝炎患者,其中AHB 12例,CHB 24例.分别用含HBV抗原C、S和P区的c18-27、s183-191和p575-583三个肽段的四聚体[Tetramer (Tc 18-27、Ts183-191、Tp 575-583)]检测患者外周血单核淋巴细胞(PBMCs)中HBV特异性CD8阳性CTL细胞的数量,同时用酶联免疫斑点法(EHSPOT)检测其分泌IFN-γ的功能.用SPSS 13.0进行统计学分析.结果 AHB和CHB患者外周血中Tc18-27特异性的CTL数量无明显不同;而Ts 183-191特异性CTL的平均值分别为0.24％±0.39％和0.03％±0.02％,阳性率分别为75％和33.3％;Tp575-583特异性CTL平均值分别为0.08％±0.09％和0.02％±0.01％,阳性率分别为50％和16.7％,AHB较CHB显著升高(P＜0.05).此外,AHB患者平均Tetramer阳性的个数为1.58个,而CHB患者平均为0.67个,AHB较CHB显著增多(P＜0.01).在9例AHB患者中,其外周血Ts183-191特异性的CTL细胞的数量为139～21 735个/106 PBMCs,用ELISPOT方法检测相对应的分泌IFN-γ的斑点形成细胞(SFC)为0～252个/106 PBMCs,AHB患者Tetramer细胞数和ELISPOT检测的IFN-γ斑点数有明显相关性(P＜0.01),而CHB患者则无此相关性.结论 AHB与CHB患者外周血中HBV特异性CTL的数量和分泌IFN-γ的差异提示CTL可能在清除病毒方面发挥着至关重要的作用,是今后慢性乙型肝炎免疫治疗的重要研究方向之一.     

乙型肝炎病毒前C/BCP区基因变异对特异性细胞毒性T淋巴细胞免疫应答的影响

目的 研究HBV前C/BCP区基因变异对慢性乙型肝炎(CHB)患者特异性细胞毒性T淋巴细胞(CTL)免疫应答的影响.方法 采用HBV核心抗原表位肽core18-27,流式细胞术胞内细胞因子(CFC)分析法检测CHB患者外周血单个核细胞(PBMC)中的特异性CTL,对扩增产物进行测序分析.结果 54例CHB患者中G1896A突变毒株21例,占38.9%;1762/1764位核苷酸联合突变26例,占48.1%;3位点同时突变毒株13例,占24.1%.3种变异株体外HBV核心抗原表位肽core18-27刺激后,特异性CTL水平[(0.41±0.09)%、(0.36±0.08)%、(0.48±0.08)%]显著高于野毒株[(0.11±0.06)%,P＜0.05].结论 G1896A变异及1762/1764位核苷酸联合突变能显著增强特异性CTL水平,在CHB患者病情活动过程中起重要作用.     

重型肝炎患者乙型肝炎病毒特异性细胞毒性T淋巴细胞应答功能研究

目的 用主要组织相容性复合物（MHC）抗原肽四聚体（Tetramer）流式细胞技术，分析乙型重型肝炎患者外周血中特异性细胞毒性T淋巴细胞（CTL）应答状况，并探讨其临床意义。方法 采用Tetramer流式细胞技术检测乙型重型肝炎患者外周血中受人类白细胞抗原Ⅰ类分子限制的三类特异性CD8^＋CTL细胞数量；采用酶联免疫吸附斑点试验技术，测定经特异性乙型肝炎病毒（HBV）肽段诱导培养的特异性CTL表达膜内细胞因子IFNγ、TNFα、IL-4和IL—10等的水平；采用Promega CytoTox96非放射性细胞毒试验技术，测定经特异性HBV肽段诱导培养的特异性CTL杀伤靶细胞能力。结果 急性乙型重型肝炎组外周血中针对HBVcore18-27表位的特异性CTL数量高于慢性乙型重型肝炎组（P〈0．05），而低于急性乙型肝炎组（P〈0．05）；急性乙型重型肝炎组表达干扰素γ和肿瘤坏死因子α较慢性乙型重型肝炎组高（P值均〈0．05）；急性乙型重型肝炎组HBVcore18—27特异性CTL裂解靶细胞能力高于慢性乙型重型肝炎组（P〈0．05）。结论 急性乙型重型肝炎患者特异性CTL应答作用增强，而慢性乙型重型肝炎患者特异性CTI。应答缺乏。急性乙型重型肝炎患者外周血特异性CTL持续存在，可能与促进病毒清除等相关。     

慢性乙型肝炎患者特异性细胞毒性T淋巴细胞的变化

目的探讨慢性乙型肝炎急性发作及病情严重程度与特异性细胞毒性T淋巴细胞(CTL)水平的关系。方法从29例人白细胞抗原(HLA)-A2阳性慢性乙型肝炎急性发作及免疫耐受慢性乙型肝炎患者外周血中分离外周血单个核细胞,用HLA-A2*HBcAg抗原表位肽-五聚体复合体及CD8单克隆抗体染色后,用流式细胞仪检测乙型肝炎病毒(HBV)特异性的CTL细胞。并对6例急性发作患者进行动态检测HBV特异性的CTL、肝功能和病毒载量。结果19例急性发作慢性乙型肝炎患者五聚体阳性细胞占CD8阳性细胞的比例平均为1．4%±0．8%,而10例免疫耐受患者为0．6%±0．4%,两组间比较,t=2．180,P＜0．05,差异有统计学意义。急性发作导致重型肝炎的患者中,乙型肝炎核心抗原特异性CTL细胞阳性率平均为1．3%±1．0%,而非重型肝炎患者为1．4%±0．8%,两组间比较,差异无统计学意义。在12周的随访期内,患者丙氨酸氨基转移酶和病毒载量逐渐下降,但乙型肝炎核心抗原特异性CTL维持在较高的水平(＞0．7%)。结论慢性乙型肝炎患者急性发作与HBV特异性CTL有关,但主要的肝细胞损害可能并非直接由HBV特异性CTL引起的。     

急性乙型肝炎患者特异性核心抗原细胞毒性T淋巴细胞频率的变化及其意义

目的 动态观察急性乙型肝炎(AHB)患者外周血HBcAg18-27表位特异性细胞毒性T淋巴细胞(CTL)、血清ALT、HBV DNA、HBsAg和淋巴细胞亚群的变化,探讨HBV特异性CTL频率的消长在病毒清除以及肝脏损伤中的作用.方法 分别选取AHB、慢性乙型肝炎(CHB)患者外周血,根据人类白细胞抗原(HLA)-A0201结果分为两组:HLA-A0201阳性患者作为HBV特异性CTL检测组、HLA-A0201阴性患者作为特异性抗原表位对照组.用HLA-A0201限制性表位HBcAg18-27五聚体复合物通过流式细胞技术,动态定量检测外周血中HBV特异性CTL频率和T、B淋巴细胞与自然杀伤细胞(NK)和NKT淋巴细胞;以速率法检测血清ALT水平;荧光定量PCR检测HBV DNA水平; Abbott微粒子化学发光技术检测HBV血清学标志物.计量资料采用均数±标准差((x)±s)表示或中位数(P25-P75)描述,组间比较采用方差分析或非参数检验(KruskalWallis检验和Mann-Whitney U检验);两种计量指标的关系采用Pearson相关分析.结果 AHB患者入院第1、2、3周外周血HBcAg表位特异性CTL频率分别为2.11%(0.20%～3.64%)、3.56%(1.05%～5.91%)、2.03%(0.33%～3.58%),高于入院第4、5、6周的0.99%(0.12%～2.16%)、0.29%(0.05%～0.76%)、0.39%(0.05%～0.46%),也显著高于CHB的0.11%(0.06%～0.29%),z值分别为-3.258,-4.041,-3.259,P值均＜0.01.AHB患者HBcAg表位特异性CTL峰值延迟于血清HBV DNA、HBsAg和ALT等指标的峰值;在AHB患者中,HBcAg表位特异性CTL高频率患者的血清HBsAg消失时间早于CTL频率较低的患者[(1.75±1.04)周与(4.33±3.51)周,t=-2.018,P＜0.05].CD3+CD8+T淋巴细胞频率的峰值出现在入院后第2周,并与HBcAg表位特异性CTL峰值相重叠,两者动态变化规律呈相关性(r=0.420,P＜0.01).AHB患者早期外周血NK、NKT淋巴细胞数量显著低于正常对照组和CHB患者,但随着病情好转而逐渐恢复,AHB患者外周血NK细胞数量变化与HBcAg特异性CTL动态变化呈负相关(r=-0.435,P＜0.01).结论 急性HBV感染者高频率的HBcAg表位特异性CTL与HBsAg的更早消失有密切关系,动态监测外周血中HBcAg特异性CTL频率变化,可以作为预测HBV感染后临床转归的参考指标;AHB患者外周血CD8+T淋巴细胞数量的变化,可以间接反应AHB患者体内HBcAg特异性CTL频率的改变.

Abstract：

Objective This report aims to investigate the dynamical changes of HBcAg18-27 epitope specific cytotoxic T lymphocytes(CTL), alanine aminotransferase (ALT), HBV DNA and HBsAg in peripheral blood of acute hepatitis B patients, and to explore the roles of HBcAg18-27-specific CTLs in virus clearance and liver injury. Methods Acute hepatitis B (AHB) and chronic hepatitis B (CHB) patients were divided into two groups according to results of HLA-A0201. Patients with positive HLA-A0201 were classified into HBcAg-specific CTL group and those with negative HLA-A0201 were referred as control group.The specific CTLs were stained with HLA-A0201 limited HBcAg18-27 epitope MHC-Pentamer and the frequencies of CTLs, T, B, NK and NKT cells were detected by flow cytometry (FCM). The serum ALT, HBV DNA and HBsAg were examined using speed analysis, quantitative PCR and abbott chemiluminescent technology. Results The frequencies of HBcAg18-27-specific CTLs in AHB patients were higher in the early three weeks as compared to the late three weeks. The apex time of HBV-specific CTL frequencies lagged behind those of HBV DNA, HBsAg and ALT. The loss of HBsAg in patients with high frequencies of HBVspecific CTL was earlier than that in patients with low frequencies (t = 2.018, P ＜ 0.05). In the second week the peak frequencies of CD3+CD8+ cells overlapped with that of HBcAg18-27-specific CTLs and with a positive correlation between (r = 0.420, P ＜ 0.05). During the early stages of AHB, the frequencies of NK and NKT cells were found significantly lower than that of control group and CHB group and the levels were back to normal after recovery. Moreover, a negative correlation existed between the frequencies of NK cells and the dynamic changes of HBcAg18-27-specific CTLs (r = -0.435, P ＜ 0.01) in AHB group. The frequencies of HBcAg18-27-specific CTLs were significantly higher as compared to CHB group in the first three weeks (z = -3.258, -4.04, and -3.259, P ＜ 0.01). Conclusion The early loss of HBsAg was closely related to the high frequencies of HBcAg18-27 specific CTLs in AHB patients. HBcAg-specific CTL frequencies in peripheral blood could be used to predict clinical outcome after HBV infection. The frequencie of CD8+ T cells can reflect the changes of frequencies of HBcAg-specific CTL. during acute HBV infection.     

慢性乙型肝炎患者外周血及肝组织中CD4+CD25+调节性T细胞和乙型肝炎病毒特异性细胞毒性T淋巴细胞的表达及意义

目的 研究CD4+CD25+调节性T细胞和HBV特异性CTL在慢性乙型肝炎患者外周血和肝组织中的表达和临床意义.方法 流式细胞分析技术和流式细胞术细胞因子测定法(CFC)检测157例HBV感染者(包括急性乙型肝炎20例、慢性乙型肝炎115例、乙型肝炎肝硬化22例)和20例健康对照组外周血和部分肝组织中CD4+CD25+调节性T细胞和HBV特异性CTL的表达.组间分析采用t检验.结果急性乙型肝炎,慢性乙型肝炎轻、中、重度患者外周血中CD4+CD25+调节性T细胞分别为(2.87±0.94)%、(3.53±1.56)%、(4.59±2.98)%和(3.65±1.73)%,明显高于对照组的(2.36±0.60)%(t值分别为2.04、5.97、3.30和3.17,P<0.01);慢性乙型肝炎轻、中、重度和乙型肝炎肝硬化患者外周血HBV特异性CTL为(0.189土0.152)%、(0.103±0.110)%、(0.118±0.120)%和(0.098±0.101)%,明显低于急性乙型肝炎患者的(0.815±0.360)%(t值分别为10.09、11.87、9.17和8.96,P<0.01).肝组织中CD4+CD25+调节性T细胞和HBV特异性CTL的表达高于外周血.结论 CD4+CD25+调节性T细胞可能通过抑制CD8+T淋巴细胞在机体抗病毒过程中发挥重要作用.     

乙型肝炎病毒感染临床转归与宿主细胞免疫的相关性研究

目的 比较慢性乙型肝炎患者、HBV携带者、HBV既往无症状感染者之间T细胞亚群和HBV特异性CD4+ T细胞应答强度的差异,分析宿主的细胞免疫状态对HBV感染后临床转归的影响,探讨乙型肝炎的发病机制,为慢性乙型肝炎的治疗提供新的线索.方法 选取2004年2-10月在北京协和医院肝炎门诊就诊的慢性乙型肝炎患者30例、HBV携带者22例、HBV既往无症状感染者9例以及正常对照11例,使用流式细胞仪检测其T细胞亚群,使用酶联免疫斑点法检测病毒特异性CD4+T细胞应答强度,分析其差异及临床意义.结果 慢性乙型肝炎组CD4+T细胞计数显著低于HBV既往无症状感染组和正常对照组;慢性乙型肝炎组、HBV携带者组、HBV既往无症状感染组病毒特异性CD4+ T细胞应答强度分别为(156±105)、(56±68)、(229±114)SFC/106PBMC(每106个外周血单个核细胞中斑点形成细胞的个数);慢性乙型肝炎组明显高于HBV携带者组(P＜0.01),而低于HBV既往无症状感染组(P＜0.05).结论 慢性乙型肝炎患者、HBV携带者、HBV既往无症状感染者病毒特异性T细胞应答强度存在差异,这种差异可能是造成HBV感染后不同临床转归的主要因素之一.     

慢性乙型肝炎患者HBV核心区与聚合酶区特异性CTL表位变异分析

目的比较急性乙型肝炎、慢性乙型肝炎与慢性重型乙型肝炎患者HBV C蛋白和Pol蛋白特异性CTL表位变异的差异,以探讨乙型肝炎重症化和慢性化的可能机理。方法对516例乙型肝炎患者的血清进行HLA-A2和A11分型;用巢式PCR扩增血清HBV C基因与Pol基因并对PCR产物进行序列测定;根据HBV S基因序列,用VirusBlast软件鉴定患者感染的HBV基因型;用Vector NTI软件对目前已知的HLA-A2限制性的4个C蛋白和5个Pol蛋白特异性CTL表位与HLA-A11限制性的1个C蛋白表位进行序列分析。结果 247例（47.86%）患者HLA-A2阳性,其中AHB 67例,CHB 109例,CSHB 71例;220例（42.64%）患者HLA-A11阳性,其中AHB 67例,CHB 107例,CSHB 46例;CTL表位变异分析结果如下：①在3组HLA-A2阳性患者,表位变异发生率无显著性差异（P〉0.05）;②在三组HLA-A2阳性HBV B基因型患者,P455-463和P816-824表位变异有极显著性差异（P〈0.01）;③在三组HLA-A2阳性HBV C基因型患者,各表位变异无显著性差异;④在三组HLA-A11阳性患者,C88-96表位变异发生率有显著性差异（P〈0.05）,三组HBV C基因型患者,表位变异发生率有极显著性差异（P〈0.01）,而在HBV B基因型患者,各表位变异无显著性差异（P〉0.05）。结论某些HBV C蛋白和Pol蛋白特异性CTL表位在AHB、CHB和CSHB患者间变异有明显差异,并且受感染病毒基因型的影响。CTL表位变异可能与乙型肝炎的重症化和慢性化机制相关。     

慢性乙型肝炎患者树突状细胞的分离培养

在我国慢性乙型肝炎是引起肝硬变和肝细胞癌最常见的病因。慢性乙型肝炎形成的机制可能是机体免疫功能低下,不能产生有效的针对HBV特异性的CTL反应,不能彻底清除HBV,使HBV病毒得以不断增殖,导致病变进行性发展。因此,治疗慢性乙型肝炎最根本的途径就是清除病毒。有研究表明,细胞免疫应答,包括辅助T细胞(Th)和细胞毒T细胞(CTL)应答,在机体清除乙型肝炎病毒(HBV)的过程中发挥重要作用,树突状细胞(dendritic cells,BC)是一种最具潜能的抗原提呈细胞,并且在增强或调节细胞介导的免疫反应中起     

热休克蛋白70-HBcAg(18～27)蛋白复合物对乙型肝炎病毒转基因小鼠的免疫研究

目的研究结核杆菌热休克蛋白(HSP)70-HBcAg(18～27))蛋白复合物对HBV转基因小鼠免疫应答的影响。方法体外构建HSP70-HBcAg(18～27)蛋白复合物,以HBV转基因小鼠为实验对象,进行体内免疫学功能研究。结果HSP70-HBcAg(18～27)蛋白复合物使HBV转基因小鼠外周血及脾中CD4~ T淋巴细胞和CD8~ T淋巴细胞大量增殖,诱导较强的抗原特异性CTL应答,并活化自然杀伤细胞、树突状细胞参与应答反应。结论HSP70-HBcAg(18～27)蛋白复合物具有免疫原性,能够增强转基因鼠细胞免疫应答能力,有望作为蛋白疫苗用于慢性乙型肝炎的免疫治疗。     

慢性乙型肝炎患者外周血树突状细胞诱导特异性T淋巴细胞应答

目的 探讨慢性乙型肝炎患者外周血树突状细胞(Dc)是否诱导特异性T细胞应答。方法(1)将研究对象分为慢性乙型肝炎患者组、急性乙型肝炎痊愈组、健康志愿者组,分离各组研究对象的外周血单个核细胞(PBMC),细胞内细胞因子染色方法检测其对细胞毒性T淋巴细胞(CTL)特异表位多肽乙型肝炎病毒核心抗原(HBcAg)18-27的记忆性免疫应答;(2)培养慢性乙型肝炎患者DC,将负载有乙型肝炎抗原表位多肽的DC诱导特异的T细胞应答。采用细胞内细胞因子染色方法检测诱导的T细胞分泌的细胞因子,乳酸脱氢酶释放法测定诱导的T细胞杀伤活性。结果(1)急性乙型肝炎患者PBMC对HBcAg 18-27 CTL特异表位多肽存在记忆的免疫应答,其分泌干扰素-γ的CD8+T细胞占CD8+T细胞总数的(4.3±2.5)％,分泌白细胞介素-2的占总细胞数的(4.8±2.2)％,分泌肿瘤坏死因子-α占总细胞数的(4.6±2.3)％。而慢性乙型肝炎患者和健康志愿者对其记忆应答很低,与急性乙型肝炎患者比较差异有显著性,t值为2.508-3.305,P<0.05。(2)用多肽共孵育过的慢性乙型肝炎患者DC多次诱导的T细胞慢性乙型肝炎患者组,加肽孵育的靶细胞比例为30:1、10:1、3:1时,其杀伤率分别为(57.0±20.3)％、(49.5±20.2)％、(21.8±12.9)％,均高于对照组,表明慢性乙型肝炎患者DC可以诱导特异的T     

A pilot study of the CY-1899 T-cell vaccine in subjects chronically infected with hepatitis B virus. The CY1899 T Cell Vaccine Study Group.

Clinical observations suggest that eradication of the hepatitis B virus (HBV) is immune-mediated. Vigorous cytotoxic T lymphocyte (CTL) activity directed at HLA class I-bound viral epitopes are detected during acute hepatitis B, but not in chronic hepatitis B carriers. A CTL epitope derived from the hepatitis B core protein amino acids 18-27 has been incorporated into a vaccine also comprised of a T-helper cell epitope and 2 palmitic acid residues (CY-1899). The aim of this study was to determine whether repeated doses of CY-1899 given to patients with chronic hepatitis B could initiate in vivo CTL activity and viral clearance. Patients with chronic hepatitis B received up to 4 doses (ranging from 0.05 mg to 15 mg) 6 weeks apart. Following vaccination, patients were monitored for hepatitis B surface antigen and "e" status, HBV-DNA levels, liver biochemistry, CTL activity, and any adverse events. Ninety patients with chronic hepatitis B infection received CY-1899. Mean CTL responses were all low but were maximal following vaccination with 5 mg CY-1899. Peak CTL responses never exceeded 10 lytic units (LU) regardless of vaccine dose, this value being well below that seen following resolution of acute hepatitis B. No significant changes in liver biochemistry or viral serology were observed during follow-up. No serious adverse events were noted. Administration of the single-epitope vaccine, CY-1899, initiated CTL activity, but of a magnitude lower than that observed during spontaneous HBV clearance. This low-level CTL activity was not associated with viral clearance.     

乙型肝炎患者外周血CD4^＋T细胞Notch1蛋白的表达

目的探讨CD4^＋T细胞Notch1蛋白在乙型肝炎病人发病机制中的作用。方法采用流式细胞仪检测23例慢性乙型肝炎患者、8例急性乙型肝炎患者和10例健康对照者的外周血CD4^＋T细胞Notch1蛋白表达水平,并应用荧光定量PCR方法测定上述研究对象血中HBV-DNA滴度。结果慢性乙型肝炎组外周血CD4^＋T细胞Notch1蛋白的平均百分率为（3.17±0.65）%,与急性乙型肝炎组（1.85±0.46）%和健康对照组（2.18±0.57）%比较都有极其显著差异（P〈0.001）;急性乙型肝炎组外周血CD4^＋T细胞Notch1蛋白的百分率与健康对照组比较差异无统计学意义（P〉0.05）;慢性乙型肝炎患者外周血CD4^＋T细胞Notch1蛋白的百分率与HBV-DNA滴度呈正相关。结论Notch1蛋白在慢性乙型肝炎患者外周血中表达增高,说明其可能参与乙型肝炎病情的发展及病毒的清除。     

Quantitative and functional analysis of core-specific T-helper cell and CTL activities in acute and chronic hepatitis B

Abstract: Aims/Background: CD4+ T-helper cell (Th) responses to hepatitis B virus (HBV) core antigen (HBc) are increased during exacerbations in acute and chronic hepatitis B (AHB, CHB) and might influence the induction of CD8+ cytotoxic T lymphocytes (CTL) that are important for viral clearance. Methods: HBc-specific proliferative responses and cytokine release of blood mononuclear cells (PBMC) were studied in patients with AHB or CHB, as well as responders and non-responders to interferon-α treatment (IFN-R, IFN-NR), by [³H]-thymidine-uptake, enzyme-linked immunosorbent assay (ELISA) and Elispot assay and were compared to peptide HBc18–27-specific CTL precursor frequencies among CD8+ T cells derived from HLA-A2+ patients. Results: HBc-specific proliferative PBMC responses and Th frequencies were significantly increased in AHB patients compared with untreated CHB patients. PBMC derived from IFN-R showed stronger cellular responses than IFN-NR. Stimulated PBMC from all patient groups secreted significantly more IFN-γ than IL-4 indicating Th1/Th0 cell responses. Furthermore, in AHB and IFN-R patients, high peptide HBc18–27-specific CTL precursor frequencies closely correlated with strong HBc-specific Th responses, whereas in untreated CHB and IFN-NR patients lower CTL frequencies were observed without correlation to Th activities. Conclusions: HBV core-specific Th-cell responses appeared to support efficient CTL induction in patients with viral clearance, whereas in chronic HBV carriers quantitatively insufficient Th and CTL responses were observed. This observation could be important for future therapeutic strategies.     

Dendritic cells from chronic hepatitis B patients can induce HBV antigen-specific T cell responses

AIM: To determine whether dendritic cells (DCs) from chronic hepatitis B patients could induce HBV antigen-specific T cell responses or not. METHODS: DCs were generated from peripheral blood mononuclear cells of patients with chronic hepatitis B (CHB) infection and healthy donors. We compared the phenotypes of these DCs and their ability to secrete cytokines and to participate in mixed lymphocyte reactions. In addition, autologous lymphocytes were cultured with DCs loaded with HBV core region peptide HBcAg8-27, an epitope recognized by cytotoxic T lymphocytes (CTL), and bearing human leucocyte antigen (HLA)-A2 for 10 d. Cytokine secretion and lytic activity against peptide-pulsed target cells were assessed. RESULTS: DCs with typical morphology were generated successfully by culturing peripheral blood mononuclear cells (PBMCs) from CHB patients with AIM-V containing GM-CSF and IL-4. Compared with DCs from normal donors, the level of CD80 expressed in DCs from CHB patients was lower, and DCs from patients had lower capacity of stimulate T cell proliferation. When PBMCs isolated from patients with chronic or acute hepatitis B infection and from normal donors were cocultured with HBcAg18-27 peptide, the antigen-specific memory response of PBMCs from acute hepatitis B patients was stronger than that of PBMCs from chronic hepatitis B patients or normal donors. PBMCs cocultured with DCs treated with HBcAg18-27 CTL epitope peptide induced an antigen-specific T cell reaction, in which the level of secreted cytokines and lytic activity were higher than those produced by memory T cells. CONCLUSION: DCs from patients with CHB can induce HBV antigen-specific T cell reactions, including secretion of cytokines essential for HBV clearance and for killing cells infected with HBV.