慢性乙型肝炎病毒感染产妇的胎儿脐带血树突状细胞的表型及功能

目的探讨孕妇慢性乙型肝炎病毒(HBV)感染对其胎儿脐血的树突状细胞(DC)的表型和功能的影响,并与健康胎儿、健康成人及成人慢性HBV感染者进行对比研究。方法体外分离慢性HBV 感染产妇及健康产妇胎儿脐带血、健康成人及慢性乙型肝炎患者外周血单个核细胞进行树突状细胞转化,行流式细胞术对其表型进行分析,通过交叉混合淋巴细胞培养和DC培养上清液白细胞介素(IL)-12检测来分析DC的功能。结果慢性HBV感染产妇脐血来源DC的CD 80、CD 83的表达显著低于健康胎儿组、健康成人组、成人慢性HBV感染组,CD14的表达则显著高于此三组,P值均<0.0 5;慢性HBV感染产妇脐带血DC分泌的IL-12水平也明显低于健康产妇、健康成人组及慢性乙型肝炎患者(P<0.05);促异体T淋巴细胞增殖能力的强弱依次为健康成人DC-健康脐血T淋巴细胞>健康成人DC-健康成人异体T淋巴细胞>健康脐血DC-异体健康脐血T淋巴细胞>健康脐血DC-健康成人T淋巴细胞>孕母HBV感染脐血DC-健康脐血T淋巴细胞>孕母HBV感染脐血DC-健康异体成人T淋巴细胞。结论慢性HBV感染产妇胎儿脐血DC的成熟和功能显著低于健康胎儿脐血DC、健康成人甚至慢性乙型肝炎患者外周血DC。     

慢性乙型肝炎患者树突状细胞功能与单个核细胞乙型肝炎病毒共价闭合环状DNA的关系

目的 探讨慢性乙型肝炎(CHB)患者外周血单个核细胞(PBMC)及树突状细胞(DC)内HBV共价闭合环状DNA(HBV cccDNA)的存在状况,DC成熟度及功能状态与DC或PBMC中HBV cccDNA载量的关系.方法 分离29例CHB患者和10例健康对照者的PBMC,用重组人粒细胞-巨噬细胞集落刺激因子(GM-CS...     

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.     

树突状细胞亚群相对数与乙型肝炎病毒复制及肝损伤的关系

目的探讨外周血树突状细胞(DC)亚群相对数量与慢性乙型肝炎(简称慢乙肝)患者血清乙型肝炎病毒(HBV)DNA水平和肝脏病理炎症损伤程度问的关系。方法定量聚合酶链反应法检测患者血清HBV DNA,利用流式细胞仪检测外周血DC亚群。结果血清HBV DNA<106拷贝/ml慢乙肝患者外周血DC2相对数量显著高于HBV DNA≥106拷贝/ml患者和健康者(P<0.05),而后两组间DC2的差异无统计学意义;上述三组中DC1相对数量差异无统计学意义;外周血DC亚群相对数与患者临床型别和肝内炎症损伤程度无关。结论外周血DC2亚群的升高与慢乙肝患者体内HBV低水平复制相关,提示DC2 可能在抑制HBV复制中发挥作用;外周血DC亚群相对数与肝组织炎症程度不相关。     

乙型肝炎病毒感染者外周血树突状细胞的检测分析

目的检测急、慢性乙型肝炎、肝硬化患者和健康人外周血树突状细胞(DC)数量和功能的变化,了解DC与乙型肝炎病毒感染(HBV)的关系。方法采用流式细胞技术检测15例急性乙型肝炎,40例慢性乙型肝炎,20例肝硬化患者及25例健康人体内DC亚群(MDC和PDC)的变化;体外分离培养产生的DC1表型及刺激混合性淋巴细胞反应(MLR)的能力反映MDC的功能;外周血单个核细胞(PBMCs)产生的α-干扰素水平反映PDC的功能。结果慢性乙型肝炎患者的DC1表面CD80、CD86、CD83的表达水平低于健康人;健康人外周血MDC的比例为0.42%±0.09%,绝对数为(12.9±6.1)×106/L,两者在慢性肝炎、肝硬化患者中均下降;而健康人外周血PDC的比例为0.37%±0.14%,绝对数为(9.0±4.1)×106/L,两者在慢性肝炎、肝硬化患者均降低,慢性肝炎和肝硬化患者PDC分泌的α-干扰素量也低于健康人。结论慢性乙型肝炎、肝硬化患者体内两种DC均出现数量和功能异常,此种异常降低了DC的抗原递呈作用,导致HBV的慢性化及疾病进程。     

儿童慢性乙型肝炎患者外周血树突状细胞亚群的变化和临床特点

目的观察慢性乙型肝炎患儿外周血树突状细胞(DC)亚群的变化,探讨DC亚群在乙型肝炎慢性化中的意义.方法慢性乙型肝炎患儿组26例,HBV携带儿童组16例,健康儿童25例,利用流式细胞技术对患者外周血髓样树突状细胞(mDC)和浆细胞样树突状细胞(pDC)的百分比和绝对值进行检测,并用CPG2216刺激外周血单个核细胞(PBMC),检测pDC的α干扰素产生能力.结果与健康对照组比较,慢性乙型肝炎患儿组血mDC和pDC的百分比和数量均显著下降(P＜0.05),PBMC经CpG2216刺激后产生干扰素的能力较健康对照组也有显著下降(P＜0.05),而HBV携带儿童仅pDC百分比与健康儿童比较有明显下降(P＜0.05).相关性分析表明,HBV携带儿童pDC绝对值与HBV DNA水平之间存在正相关(r=0.5464,P=0.0351),而慢型性乙型肝炎患儿mDC绝对值与HBV DNA水平存在负相关(r=-0.4554,P=0.0194).结论慢性乙型肝炎患儿外周血DC亚群下降的频率和数量导致其功能损伤,可能与HBV持续感染的致病机制有关.     

慢性乙型肝炎树突状细胞表型和功能的变化与免疫耐受

目的 观察慢性乙型肝炎病毒（HBV）感染者树突状细胞（DC）形态和功能的改变。方法 从13例慢性乙型肝炎患者和11例健康人外周血中分离和培养DC,观察DC的形态,用流式细胞仪检测DC表面标记HLA－DR、CD1a、CD80和CD86的表达,用^3H-TdR掺入法测定DC诱导混合性淋巴细胞反应（MLR）的能力。结果 正常DC较慢性乙型肝炎患者在形态上更为典型,不规则,HLA－DR、CD80和CD86分子的表达水平较高（P＜0.05）,诱导MLR的能力较强（P＜0.05）。结论 慢性乙型肝炎患者外周血DC处于不完全成熟状态,其免疫刺激能力较低。     

CD11c+髓样树突状细胞在慢性乙型肝炎中频数与细胞表型的变化

目的:探讨慢性乙型肝炎患者外周血中CD11c 髓样树突状细胞(mDC)相对数量和细胞表型的变化,以及与HBV持续感染之间的关系.方法:2007-03/2007-10瑞金医院感染科住院及门诊慢性乙型肝炎患者28例,另设健康对照组21例(均为本院职工).流式细胞分析技术检测受试者外周血mDC的百分比数.磁珠分选方法分离纯化mDC.流式细胞仪检测mDC表面共刺激分子CD80和CD86.结果:与健康对照组相比,慢性乙肝患者CD11c mDC占外周血单个核细胞的频数明显降低,差异有统计学意义(0.38%±0.61%vs0.77%±0.56%,P<0.05).慢性乙肝患者外周血mDc频数与血清ALT水平、病毒载量呈负相关(r=0.374,-0.435,均P<0.05),患者组不同肝脏炎症程度mDC频数存在差异.新鲜分离的mDc表面共刺激分子cD80和cD86表达较低,但患者组CD86的表达明显高于正常对照组,差异有统计学意义(45.26%±21.54%vs18.71%±10.93%,P<0.05).结论:慢性乙型肝炎患者外周血CDllc mDC亚群百分比降低,但mDc表面共刺激分子表达率并未严重受损,外周血中CD11c mDc数量减少可能与血清病毒载量及肝脏炎症程度相关.     

CpG寡核苷酸联合HBsAg对慢性乙型肝炎患者树突细胞功能及核因子活性的影响

目的 研究含CpG寡核苷酸(CpG-ODN)联合重组HBsAg对慢性乙型肝炎(CHB)患者外周血树突状细胞(DC)表型和功能及其对核因子-κB和激活蛋白-1活性的影响.方法 以重组细胞因子联合诱导扩增CHB患者外周血单个核细胞得到DC;以CpG-ODN和HBsAg及肿瘤坏死因子α刺激DC,评价其表型、功能和胞核核因子-κB、激活蛋白-1活性.结果 CpG-ODN联合HBsAg明显提高CHB患者DC表面分子人类白细胞抗原-DR表达、白细胞介素-12分泌以及刺激同种T淋巴细胞增殖的能力,尤其能提高CD1a的表达.上述免疫佐剂能增强DC细胞核因子κB活性,同时抑制激活蛋白-1活性.结论 CpG-ODN与hTNF α一样能够促进CHB患者外周血DC成熟;其作用机制可能是选择性激活或抑制DC胞内不同核因子活性.     

中药辨证干预对慢性乙型肝炎肾虚患者树突状细胞功能影响

目的研究中药辨证干预对慢性乙型肝炎（CHB）肾虚患者外周血树突状细胞（DC）功能的影响。方法 CHB肾虚证患者60例,健康对照组10例,分离各组外周血单个核细胞,体外诱导培养DC,流式细胞仪检测DC表面分子CD80、CD86、CD1a、HLA-DR表达率;酶联免疫吸附检测（ELISA）检测DC培养上清液中细胞因子IL-10表达水平,同时检测CHB患者血清HBV DNA定量并记录中医证候积分。予中药辨证干预治疗3个月,检测患者治疗后外周血DC表面分子CD80、CD86、CD1a、HLA-DR及培养上清中IL-10的表达水平及血清HBV DNA定量,并记录中医证候积分的变化。结果治疗前CHB肾虚证患者CD80、CD86、CD1a、HLA-DR表达率显著低于正常对照组（P〈0.05）,IL-10表达水平显著高于正常对照组（P〈0.05）;治疗后CHB肾虚证患者CD80、CD86、CD1a、HLA-DR表达率明显提高、IL-10表达水平明显降低（P〈0.05）,中医证候积分明显改善（P〈0.05）。结论 CHB肾虚证患者存在DC成熟障碍及功能异常,主要表现为表面共刺激分子表达率下降,抑制因子表达率升高。中药辨证干预不仅能改善患者中医证候积分,同时能提高DC表面共刺激分子表达率,降低抑制因子表达率,提示DC可能与中医虚实病机存在一定相关性。     

Occult Hepatitis B (OBH) in Clinical Settings

Context

Occult hepatitis B (OHB), or persistent HBV DNA in patients who are hepatitis B surface antigen (HBsAg) negative, is a recently recognized entity. In an attempt to summarize the issues, this review presents an overview of the current proposed hypothesis on the clinical relevance and also updates the knowledge on the classification of OHB in different clinical settings.

Evidence Acquisition

OHB could be found in different population and clinical backgrounds including: viral co-infections (with either human immunodeficiency or hepatitis C viruses), HBV chronic carriers, dialysis patients, transplantation settings and certain clinical situations (named in here: special clinical settings) with no apparent distinguishable clinical parameters.

Results

The exact magnitude, pathogenesis, and clinical relevance of OHB are unclear. Even the possible role exerted by this cryptic infection on liver disease outcome, and hepatocellular carcinoma development remains unknown.

Conclusions

Monitoring of Individuals with positive anti-HBc, mass immunization programs and improvement in diagnostic tools seem to be important to control the probability of transmission of HBV through cryptic HBV infection.     

Occult Hepatitis B Virus (HBV) Infections: Hepatitis B Surface Antigen (HBsAg) Negative and Primary Occult

Occult HBV infection (OBI) is defined as persistence of HBV genomes (with detectable or undetectable serum HBV DNA) in the liver of serum HBsAg negative individuals. It represents the HBsAg negative phase of the natural history of HBV infection in individuals with self-limited acute hepatitis B or in HBsAg carriers or chronic hepatitis B patients who lose HBsAg either naturally or after antiviral therapy and maintain lifelong anti-HBc in serum (with or without anti-HBs and\or anti-HBe). Rarely it may occur as primary “occult” infection when caused by minute viral amounts unable to induce humoral immune response. HBsAg negative infections stem from lifelong intrahepatic persistence of HBV-ccc-DNA under the host’s immune control and may lead to HBsAg positive reactivation after immunosuppressive therapies or epigenetic modifications. HBV reactivation can be avoided by pre-emptive antiviral therapy with nucleos(t)ide analogs. OBI in chronic liver disease of other etiologies may contribute to the development of hepatocellular-carcinoma.     

Seroconversion of Hepatitis B Envelope Antigen (HBeAg) by Entecavir in a Child with Chronic Hepatitis B

Hepatitis B virus (HBV) infection is a worldwide health problem. Consensus guidelines for the treatment of chronic HBV in children have not been established, and indications for antiviral therapy in adults with chronic HBV infection may not be applicable to children. The medications that are Food and Drug Administration approved for the treatment of children with HBV include interferon (IFN)-alpha and lamivudine. Nondetectable serum HBV deoxyribonucleic acid, Hepatitis B envelope antigen (HBeAg) loss, and HBeAg seroconversion following 1 year duration of entecavir treatment. A review of the literature of entecavir treatment of chronic hepatitis B in children is also provided.     

肝康栓的体外抗乙型肝炎病毒作用的研究

观察肝康栓对抗乙型肝炎病毒(HBV)体外药效评价,进一步探讨其抗病毒机制.方法:利用乙型肝炎病毒基因转染的2.2.15细胞系,用不同浓度的肝康栓作用于此细胞系,留取培养上清液.用ELISA、PCR技术检测上清液中的乙肝病毒表面抗原(HBsAg)、乙肝病毒e抗原(HBeAg)及乙肝病毒DNA(HBV DNA)的含量作为药物抗HBV效果的观察指标.结果:肝康栓能有效抑制上清液中的HBeAg和DNA浓度,但对HBsAg作用不明显.结论:肝康栓是一种有效的抗乙肝病毒的药物.     

Hepatitis B virus DNA (HBV-DNA) in anti-HBe positive sera

HBV-DNA measured by the spot hybridization technique, was found in the sera of 28 of 106 (26.4%) anti-HBe positive carriers of HBsAg. Dane particle-associated HBeAg, HBcAg and HBV-specific DNA-polymerase activity were found in the sera of nine (8.5%), five (4.7%) and two (1.9%) of these patients, respectively. All carriers with serum HBV-DNA had chronic liver disease and 18 had intrahepatic delta-Ag and serum anti-delta at titers higher than 1/5000. Intrahepatic HBcAg was detected in the nuclei of 90% of delta negative individuals; 50% of them also had cytoplasmic fluorescence. Only two of the 18 patients with intrahepatic delta-Ag (11%) had HBcAg in the liver. Viral nucleic acid was not found in the sera of 15 other patients with chronic hepatitis, seven of whom had intrahepatic delta-Ag. Serum HBV-DNA was also negative in the remaining 63 symptomless carriers of HBsAg lacking markers of delta infection. Interestingly, although DNA-polymerase negative, some sera gave autoradiographic spots of high optical density. HBV-DNA was detected in them at concentrations typical of sera which are usually both DNA-polymerase and HBeAg positive. Detection of HBV-DNA in serum represents the most direct and sensitive in vitro assay for assessing HBV infectivity and characterizes HBsAg carriers with HBV-related liver damage and ongoing HBV replication independently from the state of HBeAg/anti-HBe system. In the Mediterranean area, the majority of anti-HBe positive carriers with serum HBV-DNA have chronic liver disease and delta infection.     

Hepatitis B surface antigen (HBsAg) – fibrinogen interaction

ABSTRACT— Deparaffinized sections of hepatitis B surface antigen (HBsAg) positive liver biopsies, when incubated in a human fibrinogen solution, reveal a strong labelling of ground-glass hepatocytes with fibrinogen. This implies an interaction between HBsAg and fibrinogen. The binding of fibrinogen to HBsAg is thought to be mediated by the protein moiety of HBsAg, since its binding capacity is destroyed by trypsin digestion, and to occur through formation of disulphide bridges because the binding can be disrupted by the reducing effect of 2-mercapto-ethanol. Not all ground-glass hepatocytes exhibit fibrinogen binding. The reason for this is at present unclear. No relationship between serum HBeAg positivity and binding of HBsAg to fibrinogen was observed. Preincubation in a fibrinogen solution did not alter the immunoreactivity of HBsAg. We conclude that there is a striking analogy between the interaction of HBsAg with polymerized human serum albumin and its interaction with fibrinogen.     

扯根菜及其系统提取物抗乙型肝炎病毒体外实验研究

目的:通过体外实验,探讨扯根菜及其系统提取物抗乙肝病毒的作用。方法:将中药扯根菜及其系统提取物作用于HBVDNA转染细胞系2215细胞,通过检测细胞培养液中HBsAg、HBeAg的变化来评价扯根菜及其系统提取物抗乙肝病毒的效果及其可能的药理活性部位。结果:当水提取物浓度为264μg/ml、直接水提取物浓度为57μg/ml时,对2215细胞分泌HBeAg的抑制率分别为54.79％及54.09％,治疗指数(TI)>2。结论:扯根菜水提取物和直接水提取物达到一定浓度时,在体外有一定的抗乙肝病毒作用。