不同类型慢性乙肝病毒感染者外周血树突状前体细胞亚群的变化特点

目的观察不同类型慢性乙肝病毒感染者外周血树突状前体细胞（pDC）亚群的变化,探讨pDC亚群在慢性乙肝病毒感染自然史中的意义。方法采集健康对照组15例,免疫耐受组19例,慢性乙型肝炎组26例,非活动性组33例的外周血,用流式细胞技术检测髓样树突状前体细胞（pDC1）和浆细胞样树突状前体细胞（pDC2）。结果与健康对照组比较,慢性乙型肝炎组pDC2显著下降（P〈0.05）,慢性乙型肝炎组pDC1显著高于免疫耐受组（P〈0.05）,非活动性组pDC2显著高于免疫耐受组和慢性乙型肝炎组（P〈0.05）。相关性分析表明,HBVDNA水平与pDC2之间存在正相关（r=0.303,P=0.043）。结论慢性乙肝病毒感染自然史不同阶段的外周血树突状前体细胞（pDC）亚群的频率和数量的变化,可能与HBV持续感染的机制和肝炎的发病机制有关。     

细胞因子诱导的自体杀伤细胞回输治疗慢性乙型肝炎患者树突状细胞功能的研究

目的了解细胞因子诱导的自体杀伤（CIK）细胞回输治疗是否能恢复慢性乙型肝炎（CHB）患者树突状细胞（DC）的功能。方法12例CHB患者应用CIK细胞回输治疗,入组前6个月至CIK细胞治疗随访期间,患者未接受任何抗病毒及免疫调节治疗。观察24周后患者髓样树突状细胞（mDC）、浆细胞样树突状细胞（pDC）比例和功能变化,并对其变化结果进行分析。结果应用CIK细胞回输治疗24周后,病毒载量下降≥2个log或转阴的患者外周血mDC、pDC功能明显提高。病毒学应答患者（n=6）病毒载量下降水平与外周血DC功能的恢复密切相关。结论CHB患者接受CIK细胞回输治疗可以提高外周血mDC和pDC功能。     

慢性乙肝病毒感染者外周血树突状细胞亚群与血清HBVDNA的关系研究

目的 探讨慢性乙肝病毒感染者外周血树突状细胞(DCs)亚群[髓系DC(MDC)和浆细胞DC(PDC)]的变化与血清HBVDNA的关系.方法 采集健康人和慢性乙肝病毒感染者外周静脉抗凝全血,利用荧光抗体标记和流式细胞仪检测外周血树突状细胞亚群(MDC的特异性标记为Lineage-HLA-DR+CD11c+,PDC的特异性标记为Lineage-HLA-DR+CD123+);固相放免法定量检测血清乙型肝炎病毒标志.结果 慢性乙肝病毒感染者外周血PDC相对平均百分率低于健康对照者,但缺少统计学意义,MDC相对数量在两者间没有差别;血清HBeAg阳性慢性乙型肝炎患者外周血PDC相对数量显著高(P<0.01)于HBeAg阴性患者和健康人(P<0.05),而在上述3组中MDC相对数量无差异(P=0.194).结论 HBV感染可导致患者的免疫功能改变,使机体产生免疫耐受或DC的凋亡;慢性乙型肝炎患者外周血PDC增加与HBeAg血清转换有关,提示PDC可能有抑制病毒的作用.     

Safety and immunogenicity of hepatitis B surface antigen‐pulsed dendritic cells in patients with chronic hepatitis B

Summary. The immune modulator capacity of antigen‐pulsed dendritic cells (DC) has been documented in patients with cancers and in animal models of chronic viral infections. Cancer antigen‐pulsed DC are now used for treating patients with cancer. But viral antigen‐pulsed DC are not used in chronic viral‐infected patients because safety of antigen‐pulsed DC has not been evaluated in these patients. DC were isolated from human peripheral blood mononuclear cells by culturing with human‐grade granulocyte‐macrophage colony stimulating factor and interleukin‐4. Human blood DC were cultured with hepatitis B surface antigen (HBsAg) for 8 h to prepare HBsAg‐pulsed DC. After immunogenicity assessment of HBsAg‐pulsed DC in vitro, five million HBsAg‐pulsed DC were administered intradermally to five patients with chronic hepatitis B (CHB) 1–3 times. HBsAg‐pulsed DC were immunogenic in nature because they produced significantly higher levels of interleukin‐12 and interferon‐γ compared to unpulsed DC (P < 0.05). Also, HBsAg‐pulsed DC induced proliferation of HBsAg‐specific T lymphocytes in vitro. CHB patients injected with HBsAg‐pulsed DC did not exhibit generalized inflammation, exacerbation of liver damage, abnormal kidney function, or features of autoimmunity. Administration of HBsAg‐pulsed DC induced anti‐HBs in two patients and HBsAg‐specific cellular immunity in 1 patient. This is the first study about preparation of antigen‐pulsed DC using human consumable materials for treating patients with CHB. Because HBsAg‐pulsed DC were safe for all patients with CHB and had immune modulation capacity in some patients, phase I and phase II clinical trials with antigen‐pulsed DC in CHB and other chronic infections are warranted.     

DC-SIGN在慢性乙型肝炎病毒感染时树突状细胞成熟活化中的作用

目的研究慢性HBV感染时,DC-SIGN在树突状细胞（DC）成熟和活化中介导的作用。方法将α-甘露糖苷酶抑制剂-基夫碱作用于HepG2.2.15细胞,收获上清中的高甘露糖型HBV颗粒,并于第5 d加入到外周血单核细胞衍生的DC培养基中,培养至第7 d,采用流式细胞仪检测DC表面CDla、CD83、CD80、CD86、HLA-DR分子的表达,MTT法检测DC刺激淋巴细胞增殖的能力,ELISA法检测DC分泌IL-12的水平。结果高甘露糖型HBV组,与天然的HBV组相比,DC表面CDla、CD83、CD80、CD86、HLA-DR分子的表达增加,分泌IL-12的水平升高,刺激同种异体淋巴细胞增殖的能力亦明显增强,且上述效应均可被DC-SIGN特异性抗体所阻断。结论 DC-SIGN识别高甘露糖型HBV后可以促进DC的成熟和活化,天然的HBV可能利用α-甘露糖苷酶参与的去甘露聚糖修饰来逃避DC-SIGN的识别,从而诱导DC功能的缺陷。     

Viral suppression correlates with dendritic cell restoration in chronic hepatitis B patients with autologous cytokine‐induced killer cell transfusion

Background: Myeloid and plasmacytoid dendritic cells (mDCs, pDCs) are functionally impaired in patients with chronic hepatitis B (CHB). Adoptive immunotherapy can suppress hepatitis B virus (HBV) replication in CHB patients, but whether it can restore the functionality of mDCs and pDCs remains unknown. Methods: Autologous cytokine‐induced killer (CIK) cells obtained from 14 CHB patients were transfused back to patients, case by case, to observe the effect of CIK‐cell treatment on the frequency and functionality of mDCs and pDCs in CHB patients during a 24‐week follow‐up investigation. Results: Seven virological responders exhibited a sustained decrease in HBV load after CIK‐cell transfusion; another seven non‐virological responders showed only sustained high levels of HBV load during the 24‐week period following CIK‐cell transfusion. The rate of hepatitis B e antigen loss or seroconversion was also higher in virological responders than in non‐virological responders. Importantly, we found that the frequency and cytokine‐producing capacity of mDCs and pDCs increased significantly in virological responders, but not in non‐virological responders. In addition, these patients exhibited a close correlation between restoration DC subsets and a decrease in HBV DNA load, rather than a change in the alanine aminotransferase level. Conclusion: Cytokine‐induced killer‐cell treatment reduced HBV DNA load in some CHB patients; the efficiency at least partially correlates with the restoration of frequency and functionality of mDCs and pDCs.     

抗病毒治疗对慢性乙型肝炎患者Ⅱ型树突状细胞及淋巴细胞亚群的影响

目的分析慢性乙型肝炎(慢性乙肝)患者抗病毒治疗前后外周血Ⅱ型树突状细胞(pDC2)的数量和功能变化以及淋巴细胞亚群的数量变化。方法采用流式细胞分析技术对33例慢性乙肝患者进行抗病毒治疗前后的动态观察,拉米夫定治疗组14例,干扰素α治疗组19例。检测患者外周血pDC2的比例、数量和淋巴细胞亚群数量;用体外灭活的1型单纯疱疹病毒(HSV1)刺激外周血单个核细胞(PBMCs)并与PBMCs进行24h共培养,检测培养上清液中干扰素α的产量,以评估pDC2产生干扰素α的能力;统计分析pDC2数量、功能变化及其临床意义。结果拉米夫定组和干扰素α组患者外周血pDC2的比例、数量和产生干扰素α的功能在治疗前均低于对照组(P<0.05)。抗病毒治疗后,拉米夫定组pDC2的比例和数量有提高,伴随自然杀伤(NK)细胞及CD8+T细胞数量的提高;干扰素α组pDC2的比例无明显变化,但pDC2细胞数有提高,同时伴有CD4+T细胞、NK细胞及CD8+T细胞数量的上升。结论拉米夫定和干扰素α作用机制不同,但均可提高慢性乙肝患者的pDC2、NK细胞及CD8+细胞数量。     

清热解毒法与疏肝健脾法对慢性乙型肝炎患者外周血树突状细胞功能的影响

目的:比较清热解毒法与疏肝健脾法对慢性乙型肝炎(CHB)患者外周血树突状细胞(DCs)成熟与功能的影响。方法:采集20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(PBMCs),体外培养使DCs增殖、成熟,大鼠清热解毒和疏肝健脾药物血浆干预;检测DCs表面HLA-DR、CD-1α、CD80及CD86的表达,DCs培养上清液IL-12的水平及DCs刺激同种异体T细胞增殖能力;比较干预前后DCs功能的变化。结果:清热解毒药物血浆、疏肝健脾药物血浆干预后CD80表达率升高,且疏肝健脾组大鼠CD80表达率的提高要超过清热解毒组,而HLA-DR、CD-1α、CD86的表达水平在干预前后差异无显著性意义。两组含药血浆干预后DCs刺激同种异体淋巴细胞增殖的能力提高,但两组药物血浆之间差异无显著性意义。两组含药血浆干预前后DCs分泌IL-12水平无明显变化。结论:清热解毒法和疏肝健脾法都能促进CHB患者DCs功能的恢复,疏肝健脾法对CHB患者DCs功能的影响更为显著。     

乙型肝炎相关慢加急性肝哀竭患者外周血树突状细胞水平及临床意义

目的：探讨乙型肝炎相关的慢加急性肝衰竭（acute －on －Chronic hepatitis B liver failure,ACHBLF）患者外周血树突状细胞（DCs）水平与临床转归的关系。方法收集2010年6月－2014年9月北京佑安医院收治的 ACHBLF 患者70例,以3个月后的临床预后结局分为存活组和死亡组。多因素 Logistic 回归分析影响患者预后的临床指标,同时采用流式细胞术检测外周 DCs 亚群绝对数（107）及百分比（％）;两独立样本 t 检验或秩和检验比较两组 DCs 水平差异并通过线性相关及多因素回归分析确定其是否可作为慢加急性肝衰竭临床转归的独立预测因素。结果TBil 是影响慢加急性肝衰竭3个月预后的临床指标[比值比（OR）＝1．008,95％可信区间（95％CI）：1．003～1．014,P ＝0．003）。存活组外周血髓样 DCs（mDCs）绝对数[（7．3±6．2）×107]明显高于死亡组[（4．1±4．0）×107]（t ＝2．336,P ＜0．05）;存活组患者外周血浆系 DCs（pDCs）绝对数[（0．8±0．6）×107]也明显高于死亡组[（0．4±0．5）×107]（t ＝2．307,P ＜0．05）;mDCs 百分比（Z ＝－0．080,P ＞0．05）及 pDCs 百分比（t ＝－0．248,P ＞0．05）在两组间差异无统计学意义。mDCs 和 pDCs 绝对数与 TBil 无线性相关关系（R2值分别为0．009和0．059）,mDCs 与 pDCs 绝对数可纳入多因素回归方程,结果显示 mDCs（OR ＝0．789,95％CI：0．665～0．935,P ＝0．006）和 TBil（OR ＝1．013,95％CI：1．006～1．020,P＜0．001）是慢加急性肝衰竭预后的独立预测因素。结论ACHBLF 患者外周血 mDCs 及 pDCs 绝对数存活组高于死亡组,mDCs 绝对数是慢加急性肝衰竭预后的独立预测因素。DCs 数量的差异可能是影响肝衰竭患者临床转归的原因之一。     

替比夫定对HBeAg+慢性乙型肝炎患者外周血树突状细胞及淋巴细胞亚群的影响

目的:探讨替比夫定抗病毒治疗对HBeAg阳性慢性乙型肝炎患者细胞免疫功能的影响与HBeAg阴转的相关性.方法:治疗组,30例HBeAg阳性慢性乙型肝炎患者应用替比夫定抗病毒治疗48wk,30例健康体检者为对照组.在治疗前、治疗后12、24、48wk,对外周血DCs进行体外培养,流式细胞仪检测分析DC表面分子及淋巴细胞亚群水平.结果:30例患者抗病毒治疗48wk,26例HBVDNA阴转(86.7%HBVDNA<500copies/mL),有8例发生HBeAg阴转(26.7%).与治疗前比较,治疗12wk时DCs表面分子CD1a、CD40、CD83、CD86、MHC-D的表达无明显变化;治疗24、48wk时上调,与治疗前相比较有显著性差异(P<0.05),但均低于健康对照组.淋巴细胞亚群12wk时无明显变化;治疗24、48wk时,CD4+及CD4+/CD8+比值明显提高,与治疗前比较有显著性差异(P<0.05);CD3+T、CD8+T和CD19+B细胞未见明显改变.48wk时,HBeAg阴转患者与HBeAg未阴转患者比较,DCs表面分子CD1a、CD40、CD86,MHC-DR的表达,CD4+水平及CD4+/CD...     

Selective functional deficit in dendritic cell--T cell interaction is a crucial mechanism in chronic hepatitis B virus infection

Summary. A defect in specific T cell immunity has long been assumed to be the central mechanism of persistent Hepatitis B virus (HBV) infection. Recent studies on HBV transgenic mice have suggested, however, that functional deficit of dendritic cells (DC) was an underlying cause for the T cell dysfunction. The functions of monocyte‐derived DC were determined by studying 75 subjects that included chronic hepatitis B patients with low or high HBV load; antibody to hepatitis B surface antigen (anti‐HBs) positive individuals who had recovered completely from previous acute HBV infection; healthy donors who had received hepatitis B vaccination and were anti‐HBs positive; and immunologically naïve to HBV or the vaccine individual. Impaired interactions between monocyte‐derived DC and T cells were shown in chronic HBV infection patients, especially in those with active virus replication. The dysfunctions included: (i) failure of DC to increase human leukocyte antigen (HLA‐II), B7 expression and interleukin‐12 secretion in responses to hepatitis B surface antigen (HBsAg), (ii) defective induction of T cell proliferative response to HBsAg, (iii) failure to activate T cells to produce cytokines and (iv) deficit in the induction of antigen specific cytotoxic T lymphocytes (CTLs). In vitro treatment of DC with tumour necrosis factor‐α improved HLA‐II and B7 expression, as well as Th cell and CTL responses. It is concluded that defective DC‐T cell interactions may account for the specific T cell immune defects in chronic HBV infection. Immunotherapy that aims at restoring DC functions could offer a new opportunity for effectively managing persistent HBV infections.     

慢性乙型肝炎患者外周血中淋巴细胞亚群与肝组织炎症活动度的相关性研究

目的 探讨慢性乙型肝炎患者外周血中淋巴细胞亚群与肝组织炎症活动度的关系.方法 对36例慢性乙型肝炎患者行肝脏穿刺术,对炎症程度进行病理分级,同时应用流式细胞技术测定其外周血单个核细胞膜上相关CD分子的表达情况,并与22例健康对照组进行比较分析.结果 15例G0～G2期患者外周血中CD4 CD45RO CD45RA-及CD8 CD45RA CD62L 淋巴细胞与健康对照组相比具有统计学差异(P<0.05);14例G3期患者外周血中CD3 、CD3 CD8 、CD8 CD28 、CD8 HLA-DR CD19 及CD3-CD19 淋巴细胞与健康对照组相比具有统计学差异(P<0.05);7例G4期患者外周血CD4 CD45RA CD62L 及CD19 CD5 淋巴细胞与健康组相比具有统计学差异(P<0.05);三组间淋巴细胞数均无统计学差异.不同组别的部分淋巴细胞之间具有一定的相关关系.结论 慢性乙型肝炎患者外周血中淋巴细胞亚群数量的改变与病情的轻重有一定的相关性;不同活动期的慢性乙型肝炎患者外周血淋巴细胞之间具有一定的相关关系.     

外周血淋巴细胞亚群在慢性HBV感染过程中表达的变化分析

目的探讨HBV感染患者外周血淋巴细胞亚群在疾病进展过程中的表达变化。方法选取2018年1月-2019年4月在天津市第二人民医院住院的慢性HBV感染患者共132例,其中慢性乙型肝炎患者47例,乙型肝炎肝硬化患者44例,乙型肝炎肝硬化相关原发性肝癌患者41例。另选取同期健康体检者42例作为对照组。采用流式细胞术检测4组外周血淋巴细胞亚群精准计数,比较4组外周血淋巴细胞亚群的表达水平。正态分布的计量资料,组间方差不齐采用Welch方差分析,两两比较采用GamesHowell检验。非正态分布的计量资料多组间及进一步两两比较采用Kruskal-Wallis H检验。计数资料组间比较采用χ2检验。相关性分析采用Spearman检验。结果与对照组和慢性乙型肝炎组相比,肝硬化组和肝癌组CD3^+、CD4^+T淋巴细胞数量明显减少,差异均有统计学意义(P值均<0.05)。与对照组相比,肝癌组CD8^+T淋巴细胞数量明显减少,差异有统计学意义(P<0.05);与慢性乙型肝炎组相比,肝硬化组和肝癌组CD8^+T淋巴细胞数量明显减少,差异均有统计学意义(P值均<0.05)。与对照组和慢性乙型肝炎组相比,肝硬化组和肝癌组CD19^+B淋巴细胞数量明显减少,差异均有统计学意义(P值均<0.05)。与对照组相比,肝硬化组和肝癌组CD16^+CD56^+NK细胞数量明显减少,差异均有统计学意义(P值均<0.05);与慢性乙型肝炎组比较,肝癌组CD16^+CD56^+NK细胞数量明显减少,差异有统计学意义(P<0.05)。4组疾病进展与外周血CD3^+T淋巴细胞、CD4^+T淋巴细胞、CD8^+T淋巴细胞、CD19^+B淋巴细胞、CD16^+CD56^+NK细胞呈明显负相关(r值分别为-0.414、-0.503、-0.269、-0.435、-0.402,P值均<0.01)。结论随着疾病的进展,慢性HBV感染患者免疫状态发生变化。外周血淋巴细胞亚群精准计数能够反应机体的免疫状态,可作为慢性HBV感染临床病情演变、治疗效果及疾病预后的参考依据。     

FibroScan在乙型肝炎诊断中的应用价值

目的研究Fibroscan在乙型肝炎病毒感染患者中的应用价值;评估肝纤维化测量值的变化及临床意义。方法本研究是一个随机、开放的临床研究。2009年7月至2010年2月的74例乙型肝炎感染患者均接受至少两次Fibroscan检查,血清中ALT、AST及TBil的检查采用自动分析仪检查,所有74例患者（22例慢性乙型肝炎,32例代偿性肝硬化,20例失代偿性肝硬化）接受替比夫定治疗。结果22例慢性乙型肝炎患者FibroScan值治疗6个月后降低,在治疗前后两组FibroScan值差异有统计学意义;74例乙型肝炎病毒感染患者肝功能治疗6个月后降低,差异具有统计学意义。74例乙型肝炎病毒感染患者中,HA与LSM呈显著正相关性（r=0．517,P=0．048）。结论慢性乙型肝炎患者肝硬度经过治疗可以降低。在替比夫定治疗的慢性乙型肝炎患者中肝硬度值的变化是敏感的。FibroScan将是一个在临床新药实验中评价治疗效果的有用工具。     

Study on the function of circulating plasmacytoid dendritic cells in the immunoactive phase of patients with chronic genotype B and C HBV infection

Hepatitis B virus (HBV) infection induces a wide range of chronic liver injury. The mechanism by which HBV evades the immune surveillance system remains obscure. Plasmacytoid dendritic cells (pDCs) seem to be the major endogenous interferon (IFN)-alpha producers and represent one of the most important cell types in the regulation of antiviral innate immunity; however, the phenotype and function of pDCs in patients infected by HBV with different genotypes are yet to be determined. The aim of this study was to investigate the differences in the numbers and function of peripheral blood pDCs in the immune clearing phase of chronic HBV infection with genotypes B and C. Fifty-six patients with persistent HBV infection were included in this study, with 19 age-matched healthy subjects being used as a control group. The frequencies of pDCs were analysed by flow cytometry, and the IFN-alpha produced by peripheral blood mononuclear cells (PBMCs) after stimulation with cytidine phosphate guanosine (CpG) oligonucleotides for 24 h was determined by enzyme-linked immunosorbent assay. The genotypes of HBV were detected by polymerase chain reaction and hybridization. The results showed that the frequency and IFN-alpha-producing capacity of peripheral blood pDCs were dramatically reduced and relatively inversely correlated with the level of serum alanine aminotransferase in both groups of patients with chronic genotype B and C HBV infection. A lower reduction of IFN-alpha production by CpG-stimulated PBMCs was found in patients with genotype C than in those with genotype B in the phase of immune clearance. In conclusion, the frequency and IFN-alpha-producing capacity of peripheral blood pDCs were dramatically reduced in the immunoactive phase of chronic hepatitis B (CHB). Furthermore, the lower reduction in IFN-alpha production in patients with genotype C than in those with genotype B may correlate with the outcome of antiviral treatment in CHB patients and the progression of liver inflammation.     

Efficacy of repeated high-dose hepatitis B vaccine (80 microg) in patients with chronic liver disease

Patients with chronic liver disease (CLD) respond poorly to standard hepatitis B (HBV) vaccine given as sequential 20 microg IM shots because of an overall impaired immune response. Many of these patients go on to liver transplantation and are at risk of acquiring recurrent or de novo HBV infection. To evaluate the efficacy and safety of high-dose (80 microg) IM HBV vaccination in patients with CLD who had previously failed to respond to a standard three-dose schedule of 40 microg IM vaccine given monthly. A retrospective review was undertaken at our institution of 79 patients with CLD who were treated with high-dose (80 microg) HBV vaccinations. All had previously failed a three-dose course of 40 microg HBV vaccine. An HBV vaccine response was defined as an anti-HBs titer greater than 100 mIU/ml. Liver enzymes, creatinine, age, prothrombin time, total vaccine dose, and MELD score were recorded. No adverse events were reported. Seventy-two per cent (57/79) of the subjects had an adequate response after receiving a mean total dose of 220 mug vaccine (range 80-800 microg). Twenty-eight per cent (22/79) of the subjects did not respond after receiving a mean total dose of 420 microg vaccine (range 240-720 microg). Non-responders had more severe hepatic disease defined as a higher mean total bilirubin level (p = 0.003) and a lower mean albumin level (p < 0.05). Age, prothrombin time, MELD score, and creatinine were not statistically significant between the responders and non-responders. Repeated high-dose (80 microg) HBV vaccination, in patients who do not respond to standard HBV vaccine doses, is safe and effective in the majority of patients with CLD.     

Prevalence of hepatitis delta virus (HDV) infection in chronic hepatitis B patients in eastern Turkey: still a serious problem to consider

Summary. Hepatitis delta virus (HDV) is a serious cause of liver‐related morbidity and mortality. Coexistent infection with HDV tends to aggravate the course of hepatitis B virus (HBV)‐associated liver disease. The aim of this study was to determine the prevalence of HDV infection among patients chronically infected with HBV in the Elazig region, which is in eastern Turkey. A group of 282 patients infected with chronic HBV were investigated for the study. Anti‐HDV seropositivity was evaluated in all patients. The anti‐HDV‐positive patients were further tested for HDV RNA. Severity of liver disease was assessed by liver biopsy. Regression analysis was used to determine the relationship between independent variables and HDV positivity. Of 282 chronic HBV patients, 192 were men (68.1%) and 90 were women (31.9%). The mean age was 43.8 ± 12.7 (between 18 and 73 years). Anti‐HDV was positive in 45.5% of the patients (128/282). Among the 128 anti‐HDV‐positive patients, 116 were checked for HDV RNA and 56.9% were found positive (66/116). Chronic HDV infection rate was therefore present in at least 23.4% of the whole study group (66/282). There were 83 patients with cirrhosis (29.4%) in the study group. Anti‐HDV seroprevalence and HDV RNA presence were higher in those with cirrhosis (61.4% and 42.2%, respectively). No significant relationship was found between anti‐HDV seropositivity and demographic factors such as age, sex and operation or transfusion history except family history. HDV‐RNA‐positive patients had significantly higher ALT and lower albumin levels when compared to HDV‐RNA‐negative patients. HDV‐RNA‐positive patients also had a significantly higher fibrosis stage. In conclusion, these findings demonstrated that HDV infection is endemic and still a serious problem in the Elazig region of eastern Turkey. HDV infection is significantly related to the family exposure and increases the risk of severe liver fibrosis in this region.     

慢性乙型肝炎患者外周血淋巴细胞亚群与病程相关性的研究

目的对慢性乙型肝炎轻中度、重度和肝硬化患者外周血淋巴细胞亚群的百分比和绝对细胞数进行观察,探讨慢性乙型肝炎患者外周血淋巴细胞亚群的变化与病程的关系.方法采集88例慢性乙型肝炎患者柠檬酸钠新鲜抗凝血,经流式细胞仪进行免疫分型检测.结果慢性乙型肝炎重度患者的CD3+CD4+细胞百分比显著低于轻中度患者(P＜0.05),肝硬化患者的CD3+和CD3+CD8+细胞百分比显著低于轻中度患者(P＜0.01).肝硬化患者CD3CD19+细胞百分比显著高于重度和轻中度患者(P＜0.01).CD4/CD8比例在慢性乙型肝炎轻中度、重度和肝硬化患者间无显著差异.肝硬化和重度患者淋巴细胞、CD3+、CD3+CD4+、CD3+CD8+细胞的绝对细胞数均显著低于轻中度患者(P＜0.01),且肝硬化患者CD3-CD16+56+细胞的绝对细胞数显著低于轻中度患者(P＜0.05).肝硬化患者与轻中度患者的DNA载量分布差异有显著性(P＜0.01),其高水平病毒载量的患者比例高于轻中度患者.结论慢性乙型肝炎轻中度发展为重度和肝硬化的过程中,外周血淋巴细胞亚群绝对细胞数随病情的进展显著减少,主要表现为CD3+CD4+和CD3+CD8+的T淋巴细胞亚群的百分比进行性降低.     

HBsAg loading on dendritic cells in patients with chronic hepatitis B: expressions of phenotypic molecules

BACKGROUND: The antigen reducing ability of dentritic cells (DCs), a kind of antigen presenting cells (APCs) initiating immune response, is associated with the specific immune tolerance of chronic hepatitis B(CHB) patients. However, the dysfunction of DCs can be possibly reversed by the stimulation of antigen peptides. In this study, DCs were cultured from peripheral blood monocytes (PBMCs) in patients with CHB in vitro, and the expression of phenotypic molecules on DCs loaded by different concentrations of HBsAg was observed. METHODS: Forty patients with CHB were divided randomly into 4 groups(10 patients in each group). PBMCs were isolated, and DCs were cultured after addition of granulocyte/macrophage colony-stimulating factor (GMCSF) and interleukin 4(IL-4). On the 9th day, DCs of the experimental groups were loaded at HBsAg concentrations of 2.5mg/L, 5mg/L and 10mg/L for 24 hours, whereas those of the control group were not loaded. An electron microscope was used to analyze the morphological changes of the DCs. The expression of phenotypic molecules on DCs in different groups was detected with flow cytometry. RESULTS: A combination of GM-CSF and IL-4 produced DCs from PBMCs in patients with CHB after being cultured for 9 days, whose morphological changes were tested by an electron microscope. The expression of phenotypic molecules on DCs in the control group was as low as CD83 (8.02±3.99)%, CD80(8.77±2.06)%, and MHC-DR (14.05±2.66)%. Loaded by different concentrations of HBsAg, the up-regulation of phenotypic molecules on DCs was found, with CD83(18.35±2.93)%, CD80(42.63±7.15)% and MHC-DR(47.49±6.59)% in 2.5mg/L HBsAg loading group, CD83(17.88±3.12)%, CD80(45.24± 10.93)% and MHC-DR(47.07±8.52)% in 5mg/L HBsAg loading group and CD83(16.74±2.86)%, CD80(44.59±6.99)% and MHC-DR(48.59±7.42)% in 10mg/L HBsAg loading group, respectively. Compared with the control group, the phenotypic molecules in the experimental groups were all different significantly (P<0.01), but among them, there were no differences (P>0.05). CONCLUSIONS: DCs cultured from PBMCs in the patients with CHB under the conditions of GM-CSF and IL-4 present on the typical dendritic morphology but are immature for expressing low phenotypic molecules. Loaded by different concentrations of HBsAg, the immature DCs can differentiate to mature DCs for expressing increasing phenotypic molecules.