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.     

PD-1表达对急性乙型肝炎患者HBV特异性CD8＋T细胞功能的影响

目的 观察急性乙型肝炎发病过程中患者体内病毒特异性CD8^＋细胞（CTL）上免疫抑制性分子程序性细胞死亡受体1（PD-1）的表达及临床意义。方法合成4种识别HBV不同抗原表位的五聚体，同时采集16例HLA-A2阳性急性乙型肝炎患者外周血，用五聚体染色后，通过流式细胞仪分析病毒特异性CTL细胞的频率及其表面PD-1分子的表达特点。同时进行肝功能、乙型肝炎病毒（HBV）血清标志物和血清HBVDNA检测。结果所有急性乙型肝炎患者发病早期均表现出多特异性的CTL反应，且PD-1在这些细胞上表达明显上调，显著高于来自同一患者的巨细胞病毒（CMV）和Flu特异性CTL上PD-1分子的表达水平。体外阻断PD-1／PD-L1间的相互作用能够恢复HBV特异性CTL分泌细胞因子的能力。相关性分析显示PD-1分子的上调表达与转氨酶水平正相关。结论在HBV急性感染过程中，尤其是在HBV基本被清除后，HBV特异性CTL细胞PD-1分子表达明显升高，有效抑制病毒特异性CTL分泌IFN-γ的能力，后者可能有利于避免急性乙型肝炎患者肝脏遭受更为严重的免疫损伤。     

急性乙型肝炎患者体内巨细胞病毒特异性CD8 T细胞漂移的研究

目的通过观察急性乙型肝炎发病早期患者体内巨细胞病毒（CMV）CTL的数量和程序性细胞死亡受体1（PD-1）的表达水平,探讨该信号途径对CMVCTL频率和功能的影响。方法合成识别CMVV抗原表位的五聚体,同时采集12例HLA-A2阳性的急性乙型肝炎患者的外周血,用五聚体染色后,通过流式细胞仪分析CMVPP65表位特异性CTL的频率及其表面PD-1分子的表达特点,通过阻断PD-1/PD-L1信号途径的方法,探讨PD-1分子的表达对CMVCTL功能的影响。同时进行肝功能、HBsAg、抗-HBs和血清HBV DNA载量检测。结果急性乙型肝炎发病早期,患者体内CMVCTL的频率显著低于健康对照（P〈0.001）,同时PD-1分子的表达显著升高（P〈0.05）,该分子的上调表达与CMV特异性CD8T细胞的凋亡有关,而且阻断PD-1分子与其配体PD-L1的结合,能够有效恢复CMV特异性CD8T细胞分泌因子IFN-γ的能力。结论急性HBV感染造成异质性病毒CMVCTL的数量减少和功能降低,且免疫抑制性分子PD-1参与并调节了CMVCTL的数量减少和功能的降低,提示在人的病毒感染早期,免疫抑制性信号途径参与的删除前存在的记忆性CD8T细胞会为抵抗新病毒感染的抗原特异性T细胞提供更多的空间。     

慢性乙型肝炎患者抗病毒治疗早期T细胞表面PD-1的表达

背景：慢性乙型肝炎病毒（HBV）感染者T细胞表面程序性死亡受体1（PD-1）呈持续性高表达。然而关于抗病毒治疗前后慢性乙型肝炎（CHB）患者T细胞表面PD-1表达变化及其与病毒载量关系的报道较少。目的：动态观察CHB患者抗病毒治疗早期外周血CD4＋和CD8＋T细胞表面PD-1表达水平,探讨其表达与血清HBV DNA载量和丙氨酸氨基转移酶（ALT）水平之间的关系。方法：以流式细胞术分别检测31例CHB患者抗病毒治疗前或基线期（T1）、治疗后4～8周（他）和12．16周（T3）的外周血CD4^＋和CD8^＋T细胞表面PD-1表达水平,以实时荧光定量聚合酶链反应（PCR）检测血清HBV DNA载量,同时检测血清ALT水平。结果：抗病毒治疗早期,CHB患者外周血CD4^＋和CD8^＋T细胞表面PD．1表达水平逐渐下调（P〈0．05）,血清HBV DNA载量和ALT水平亦逐步降低（P〈0．01）;CD4^＋和CD8^＋T细胞表面PD-1表达与HBV DNA载量（P〈0．01）和ALT水平（P〈0．05）均呈正相关。结论：有效的抗病毒治疗可通过降低CHB患者的病毒载量使T细胞表面PD-1表达下调,T细胞表面PD-1表达水平与患者疾病状态密切相关。     

HBcAg特异性CTL在急性乙型肝炎与慢性乙型肝炎急性发作患者外周血中的数量差异

目的:研究急性乙型肝炎(AHB)与慢性乙型肝炎(CHB)急性发作患者外周血中HBcAg特异性细胞毒性T淋巴细胞(CTL)的数量差异.方法:选择HLA-A2阳性的AHB患者17例和CHB患者13例作为研究对象,应用Tetramer流式细胞技术检测其外周血单个核细胞(PBMC)中的HBcAg特异性CTL细胞频率.结果:30份PBMC标本中测得HBcAg特异性CTL为0.071-31.610,而阴性对照的健康者PBMC标本检测值仅为0.003.AHB患者的HBcAg特异性CTL显著高于CHB患者(9.601±10.425vs0.259±0.193,P=0.002).结论:外周血中HBcAg特异性CTL的数量在AHB与CHB急性发作患者之间存在显著性差异,有望成为早期鉴别上述两种疾病状态的实验方法.     

免疫清除期慢性乙型肝炎患者外周血T细胞程序性坏死因子-1表达及其意义

目的 明确程度性坏死因子（PD-1）在免疫清除期慢性乙型肝炎患者外周血T细胞表达状态及其对患者病毒载量水平和生化指标的影响.方法 45例ALT升高的慢性乙型肝炎患者被纳入本研究.应用流式细胞术对所有患者的外周血总CD8+T细胞和CD4+T细胞PD-1表达百分比和表达强度进行检测,其中18例患者接受肝组织活检,并应用免疫...     

慢性HBV感染者外周血CD8+T细胞中Mg~(2+)水平及其临床意义

目的检测慢性乙型肝炎病毒(HBV)感染者外周血CD8+T细胞中Mg~(2+)水平以及其表面活性抑制分子PD-1、CD95和活化分子CD38、NKG2D的表达情况,为HBV的临床治疗提供理论依据。方法选取2016年1月至2017年2月在我院肝病门诊就诊的未经抗病毒治疗的HBV感染患者62例及健康对照者30例为研究对象,根据外周血HBV DNA水平将HBV感染患者分为高病毒载量组和低病毒载量组。检测HBV感染患者及健康对照者外周血及CD8+T细胞内Mg~(2+)浓度,采用流式细胞术检测三组对象CD8+T细胞表面分子PD-1、CD95、CD38和NKG2D的表达水平。结果 HBV感染患者及健康对照者外周血Mg~(2+)浓度比较差异无统计学意义(P0.05);慢性HBV感染者CD8+T细胞中Mg~(2+)浓度显著低于健康对照者(P0.05),高病毒载量组患者CD8+T细胞中Mg~(2+)浓度显著低于低病毒载量组(P0.05)。HBV感染患者CD8+T细胞表面分子PD-1表达水平显著高于健康对照者,高病毒载量组患者PD-1的表达水平显著高于低病毒载量组(P0.05);高病毒载量组患者CD95表达水平显著高于低病毒载量组和健康对照组(P0.05),低病毒载量组与健康对照组CD95表达水平比较差异无统计学意义(P0.05);高病毒载量组患者CD38表达水平显著低于低病毒载量组和健康对照组(P0.05),低病毒载量组与健康对照组CD38表达水平比较差异无统计学意义(P0.05)。HBV感染患者NKG2D表达水平显著低于健康对照者(P0.05),高病毒载量组患者NKG2D表达水平显著低于低病毒载量组(P0.05)。结论 HBV感染者外周血CD8+T细胞内Mg~(2+)浓度下降可能与CD8+T细胞功能耗竭存在一定的关联,可能与Mg~(2+)转运体功能异常有关。     

活动性慢性乙型肝炎患者特异性 CD8＋T 细胞转录因子 T-bet 表达及临床意义

目的：分析转录因子 T-bet 在活动性慢性乙型肝炎患者外周血 HBV 特异性 CD8＋ T 细胞中的表达及其临床意义。方法用 HBV 肽五聚体结合流式细胞仪检测 HLA-A2阳性的活动性慢性乙型肝炎患者和急性乙型肝炎患者外周血 HBV 特异性 CD8＋ T 细胞中 T-bet 的表达。比较分析 HBV 特异性 CD8＋ T 细胞中 T-bet 表达在急性、慢性乙型肝炎之间的差异,并进一步分析慢性乙型肝炎患者 HBV 特异性 CD8＋ T 细胞中 T-bet 表达与 HBV DNA 水平、ALT 水平以及临床预后之间的相关性。结果急性乙型肝炎患者外周血 HBV 特异性 CD8＋ T 细胞中 T-bet 表达水平显著高于活动性慢性乙型肝炎患者。活动性慢性乙型肝炎患者及 CHB 患者外周血 HBV 特异性 CD8＋ T 细胞中 T-bet 表达水平与 HBV DNA 水平及 ALT 水平无明显相关性,HBV 特异性 CD8＋ T 细胞中 T-bet 表达水平高的活动性慢性乙型肝炎患者发生HBeAg 血清转换率显著升高。结论活动性慢性乙型肝炎患者特异性 CD8＋ T 细胞转录因子 T-bet 表达与 HBV 感染的免疫控制密切相关。     

急性乙型肝炎患者自然杀伤细胞的变化与疾病进展的关系

目的 探讨急性乙型肝炎(acute hepatitisB,AHB)患者外周血中自然杀伤细胞(natural killer cells,NK细胞)频率、功能变化及与疾病进展的关系.方法 入组15例AHB患者(AHB组)和14名健康者(健康对照组),采集外周血,用流式细胞仪分析NK细胞频率和活化指标,并与临床病毒学和生化指标进行相关性分析,同时观察NK细胞对K562细胞刺激的应答.结果 AHB组NK细胞频率低于健康对照组(P＜0.05).与健康对照组相比,AHB组NK细胞呈现高活化状态,且与转氨酶呈正相关.进一步研究发现,AHB组NK细胞分泌细胞因子IFNγ及脱颗粒活性(CD107a)的能力显著增强.结论 AHB组NK细胞高活化、分泌细胞因子能力和细胞毒能力显著增强,与临床指标有相关性,提示NK细胞在加速病毒清除的同时可引起肝损伤.     

乙型肝炎患者外周血CD4~+T细胞Runx3的表达及意义

目的探讨乙型肝炎患者外周血CD4+T细胞Runx3 mRNA的表达及意义。方法应用实时定量PCR检测37例慢性乙型肝炎(chronic hepatitis B,CHB)患者(CHB组),11例急性乙型肝炎(acute hepatitis B,AHB)患者(AHB组)和19名健康对照者(健康对照组)外周血CD4+T细胞Runx3 mRNA的表达水平。结果 CHB组Runx3 mRNA的表达水平明显低于AHB组和健康对照组(P均0.01)。AHB组Runx3 mRNA的表达水平明显高于健康对照组(P0.05)。CHB重度组Runx3 mRNA的表达水平明显高于CHB轻度组(P0.05)。结论 Runx3在CHB的发病过程中可能发挥了重要的作用,CHB患者Runx3表达水平降低可能与其Th1/Th2失衡有关。     

Longitudinal fluctuations in PD1 and PD-L1 expression in association with changes in anti-viral immune response in chronic hepatitis B

ABSTRACT: BACKGROUND: Controversy exists regarding the role of PD1 and its ligand PD-L1 in chronic hepatitis B infection. In some studies, persistent HBV infection has been attributed to high levels of PD-1 and PD-L1 expression on HBV-specific T-cells and antigen-presenting cells (APCs) respectively. Other studies revealed that the up-regulation of PD-1 and PD-L1 during an acute inflammation phase is required to offset increasing positive co-stimulatory signals to avoid severe damage by an over-vigorous immune response. METHODS: Fifteen chronic hepatitis B patients, with inflammatory flare episode, were recruited prospectively. Based on serum HBV-DNA, HBsAg load, and ALT values, inflammatory flare episode were divided into initial, climax, decline and regression phase. Blood sample and liver biopsy tissues from each individual were taken in these 4 phases respectively. Circulating and intra-hepatic PD1 and PD-L1 expression levels were monitored throughout the inflammatory flare episode by flow cytometry and immunostaining and these expression levels were related to the HBV-specific T-cell changes, expression of pro-inflammatory cytokines, HBV-DNA replication and HBV antigen load. RESULTS: ]The levels of PD-1 and PD-L1 expressions were significantly up-regulated in the inflammation ascending phase, initial and climax period and in parallel with HBV-specific colon expansion. It showed increasing the level of serum ALT and decreasing the HBV-DNA loads. As the level of inflammation reduced, the circulating and intra-hepatic PD1 and circulating PD-L1 decreased progressively in concordance with serum ALT, HBV-DNA and HBsAg loads decreased except intra-hepatic PD-1 expression. Intra-hepatic PD-L1 expression did not decrease significantly during the regression phase of inflammation compared to that in prior period. The intra-hepatic PD-L1 expression remained relatively on higher level when serum HBV-DNA load and ALT decreased to approximately normal range. CONCLUSION: The relatively high level of intra-hepatic PD-L1 expression during the inflammatory regression period may contribute to constitute an immunosuppressive microenvironment, which facilitate persistent HBV infection via the inhibition of HBV-specific T cell clonal expansion.     

Programmed death-1 expression is associated with the disease status in hepatitis B virus infection

AIM： TO define the potential role of programmed death-i/programmed death-ligand （PD-1/PD-L） pathway in different hepatitis B virus （HBV） infection disease status; we examined the expression of PD-1 on antigen specific CD8＋T cells in peripheral blood of patients with chronic hepatitis B （CriB） and acute exacerbation of hepatitis B （AEHB） infection.
METHODS： The PD-1 level on CD8＋ T lymphocytes and the number of HBV specific CD8＋ T lymphocytes in patients and healthy controls （HCs） were analyzed by staining with pentameric peptide-human leukocyte antigen2 （HLA2） complexes combined with flow cytometry. Real-time quantitative polymerase chain reaction （PCR） was used to measure the serum HBV- DNA levels.
RESULTS： The level of PD-1 expression on total CD8＋ T cells in CHB patients （13.86% ± 3.38%） was significantly higher than that in AEHB patients （6.80%± 2.19%, P 〈 0.01） and healthy individuals （4.63% ± 1.23%, P 〈 0.01）. Compared to AEHB patients （0.81% ± 0.73%）, lower frequency of HBV-specific CD8＋ T cells was detected in chronic hepatitis B patients （0.37% ± 0.43%, P 〈 0.05）. There was an inverse correlation between the strength of HBV-specific CD8＋ T-cell response and the level of PD-1 expression. Besides, there was a significant positive correlation between HBV viral load and the percentage of PD-1 expression on CD8＋ T cells in CriB and AEHB subjects （R = 0.541, P 〈 0.01）. However, PD-1 expression was not associated with disease flare-ups as indicated by alanine aminotransferase （ALT） levels （R = 0.066, P 〉 0.05）.
CONCLUSION： Our results confirm previous reports that HBV specific CD8＋T-cell response in the peripheral blood is more intense in patients with AEHB than in chronic hepatitis B wlth persistent viral infection. Moreover, there is a negative correlation between the level of PD-1 and the intensity of virus specific CD8＋ T cell response.     

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

目的 研究程序性死亡分子-1(PD-1)及其配体(PD-L1)表达水平与慢性HBV感染者HBV DNA水平的相关性及抗病毒治疗对其表达的影响.方法 检测137例慢性HBV感染者的外周血单个核细胞(PBMC)表面PD-1和PD-L1,并检测其中64例人类白细胞抗原(HLA)-A2阳性者HBV特异性CTL数量.ELlSA法检测PBMC体外培养上清液中IFN-γ浓度.比较10例HBeAg阳性慢性乙型肝炎(CHB)患者予替比夫定抗病毒治疗24周前后上述指标的变化.两组间均数比较采用两独立样本的t检验,多组间的差异采用单因素方差分析,相关分析采用Pearson相关分析.结果 HBV DNA＜3 lg、3～6 lg和＞6 lg拷贝/mL组问PBMC表面PD-1和PD-L1表达均明显高于健康对照组,但差异无统计学意义;3组HBV特异性CTL表面PD-1表达分别为(69.3±11.2)%、(76.5±9.1)%和(78.0±11.7)%,HBV DNA＞6 lg拷贝/mL 组PD-1表达明显高于＜3 lg拷贝/mL组,而HBV特异性CTL数量明显低于＜3 lg拷贝/mL组;3组PBMC体外培养上清液中IFN-γ水平差异无统计学意义.HBeAg阳性组和阴性组间上述指标差异无统计学意义.替比夫定抗病毒治疗12周和24周时,PD-1、PD-L1表达较治疗前明显下降,伴有HBV特异性CTL数量逐渐增加和IFN-γ水平升高.结论 慢性HBV感染者PBMC表面PD-1的表达较健康者明显上调,且HBV特异性CTL表面表达PD-1水平与血清HBV DNA水平相关,但与HBeAg状态无关.抑制HBV复制能降低PD-1、PD-L1表达,并增加HBV特异性CTL的数量和功能.

Abstract：

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.     

The Expression of Programmed Death-1 in Circulating CD4+ and CD8+ T Cells during Hepatitis B Virus Infection Progression and Its Correlation with Clinical Baseline Characteristics

Background/Aims

Programmed death-1 (PD-1) expression was investigated in CD4⁺ and CD8⁺ T cells from hepatitis B virus (HBV)-infected patients at the chronic hepatitis B (CHB) infection, liver cirrhosis (LC), and hepatocellular carcinoma (HCC) stages.

Methods

PD-1 expression in circulating CD4⁺ and CD8⁺ T cells was detected by flow cytometry. The correlations between PD-1 expression and HBV viral load, alanine aminotransaminase (ALT) levels and aspartate aminotransferase (AST) levels were analyzed using GraphPad Prism 5.0.

Results

PD-1 expression in CD4⁺ and CD8⁺ T cells was significantly increased in both the CHB group and advanced-stage group (LC plus HCC). In the CHB group, PD-1 expression in both CD4⁺ and CD8⁺ T cells was positively correlated with the HBV viral load, ALT, and AST levels. However, in the LC plus HCC group, significant correlations between PD-1 expression and the clinical parameters were nearly absent.

Conclusions

PD-1 expression in peripheral CD4⁺ and CD8⁺ T cells is dynamic, changes with HBV infection progression, and is related to HBV viral load and liver function, especially in CHB. PD-1 expression could be utilized as a potential clinical indicator to determine the extent of virus replication and liver injury.     

Foxp3+ regulatory T cells protect the liver from immune damage and compromise virus control during acute experimental hepatitis B virus infection in mice

The strength of antiviral T cell responses correlates with clearance of hepatitis B virus (HBV) infection, but the immunological mechanisms mitigating or suppressing HBV-specific T cells are still poorly understood. In this study, we examined the role of CD4(+) Foxp3(+) regulatory T cells (Tregs) in a mouse model of acute HBV infection. We initiated HBV infection via an adenoviral vector transferring a 1.3-fold overlength HBV genome (AdHBV) into transgenic DEREG mice, where Tregs can be transiently but selectively depleted by injection of diphtheria toxin. The effect of Treg depletion on the outcome of HBV infection was characterized by detailed virological, immunological, and histopathological analysis. Numbers of Tregs increase in the liver rapidly after initiation of HBV replication. Initial depletion of Tregs revealed their complex regulatory function during acute infection. Tregs mitigated immunomediated liver damage by down-regulating the antiviral activity of effector T cells by limiting cytokine production and cytotoxicity, but did not influence development of HBV-specific CD8 T cells or development of memory T cells. Furthermore, Tregs controlled the recruitment of innate immune cells such as macrophages and dendritic cells to the infected liver. As a consequence, Tregs significantly delayed clearance of HBV from blood and infected hepatocytes. Conclusion: Tregs limit immunomediated liver damage early after an acute infection of the liver, thereby contributing to conservation of tissue integrity and organ function at the cost of prolonging virus clearance. (HEPATOLOGY 2012;56:873-883).     

Kinetics of peripheral hepatitis B virus-specific CD8+ T cells in patients with onset of viral reactivation

Background

Patients with resolved hepatitis B virus (HBV) infection undergoing chemotherapy or immunosuppressive therapy are potentially at risk of HBV reactivation. However, it remains unclear how liver disease develops after HBV reactivation. To compare the host immune response against HBV, we performed immunological analyses of six HBV reactivation patients.

Methods

The numbers of peripheral HBV-specific CD8⁺ T cells were investigated longitudinally in six HLA-A2- and/or A24-positive patients with HBV reactivation. In addition, 34 patients with resolved HBV, 17 patients with inactive chronic hepatitis B (ICHB), 17 patients with chronic hepatitis B (CHB) and 12 healthy controls were analyzed. The number and function of HBV-specific CD8⁺ T cells were assessed by flow cytometry using tetramer staining and intracellular IFN-γ production. Furthermore, the numbers of CD4⁺CD25⁺ or CD4⁺Foxp3⁺ T cells and serum inflammatory cytokine levels were analyzed.

Results

The frequency of HBV-specific CD8⁺ T cells was significantly increased in HBV reactivation patients compared with ICHB and CHB patients. In addition, the number of HBV-specific CD8⁺ T cells was increased in resolved HBV patients compared with ICHB patients. PD-1 expression was decreased in HBV reactivation patients compared with ICHB and CHB patients. The numbers of HBV-specific CD8⁺ T cells and CD4⁺CD25⁺ or CD4⁺Foxp3⁺ T cells were negatively correlated following onset of HBV reactivation.

Conclusions

During HBV reactivation, the frequency of HBV-specific CD8⁺ T cells increased even though the administration of immunosuppressive drugs and interactions with CD4⁺ regulatory T cells may be important for the onset of liver disease.