HBV抗原定量对干扰素联合阿德福韦酯治疗HBeAg阳性慢乙肝的疗效预测

Objective To investigate the levels of HBsAg in predicting the efficacy of peglated interferon-alpha 2a combined with adefovir dipivoxil( ADV ), in HBeAg-positive chronic hepatitis B patients.Methods This trial enrolled 62 HBeAg-positive chronic hepatitis B patients with detectable HBsAg for at least 6 months prior to screening, serum HBV DNA levels of at least 100 000 IU/ml. The efficacy assessment: viral suppression below 100 IU/ml. The patients with HBV DNA ≤ 100 IU/ml after 24 weeks therapy were divided into group A, in which monotherapy continued; While the rest were divided into group B, in which ADV was combined until week 48. In group B, at the end-of-treatment, the patients with HBV DNA ≤100 IU/ml were divided into group B1, the rest were divided into group B2. Results There was no significant difference on the beseline characteristics of patients between B1 and B2. There was significant difference on the levels of HBsAg at 12-week and 24-week between B1 and B2; while there was no significant difference on the levels of HBeAg. Conclusions The levels of HBsAg at 12-week and 24-week would be predictors to evaluate the efficacy of combined therapy in HBeAg-positive chronic hepatitis B patients.     

HBV抗原定量对干扰素联合阿德福韦酯治疗HBeAg阳性慢乙肝的疗效预测

Objective To investigate the levels of HBsAg in predicting the efficacy of peglated interferon-alpha 2a combined with adefovir dipivoxil( ADV ), in HBeAg-positive chronic hepatitis B patients.Methods This trial enrolled 62 HBeAg-positive chronic hepatitis B patients with detectable HBsAg for at least 6 months prior to screening, serum HBV DNA levels of at least 100 000 IU/ml. The efficacy assessment: viral suppression below 100 IU/ml. The patients with HBV DNA ≤ 100 IU/ml after 24 weeks therapy were divided into group A, in which monotherapy continued; While the rest were divided into group B, in which ADV was combined until week 48. In group B, at the end-of-treatment, the patients with HBV DNA ≤100 IU/ml were divided into group B1, the rest were divided into group B2. Results There was no significant difference on the beseline characteristics of patients between B1 and B2. There was significant difference on the levels of HBsAg at 12-week and 24-week between B1 and B2; while there was no significant difference on the levels of HBeAg. Conclusions The levels of HBsAg at 12-week and 24-week would be predictors to evaluate the efficacy of combined therapy in HBeAg-positive chronic hepatitis B patients.     

Blockade of Tim-3 Pathway Ameliorates Interferon-γ Production from Hepatic CD8^＋ T Cells in a Mouse Model of Hepatitis B Virus Infection

T cell immunoglobulin- and mucin-domain-containing molecule-3 （Tim-3） has been reported to participate in the pathogenesis of inflammatory diseases. However, whether Tim-3 is involved in hepatitis B virus （HBV） infection remains unknown. Here, we studied the expression and function of Tim-3 in a hydrodynamics-based mouse model of HBV infection. A significant increase of Tim-3 expression on hepatic T lymphocytes, especially on CD8^＋ T cells, was demonstrated in HBV model mice from day 7 to day 18. After Tim-3 knockdown by specific shRNAs, significantly increased IFN-γ production from hepatic CD8^＋ T cells in HBV model mice was observed. Very interestingly, we found Tim-3 expression on CD8^＋ T cells was higher in HBV model mice with higher serum anti-HBs production. Moreover, Tim-3 knockdown influenced anti-HBs production in vivo. Collectively, our data suggested that Tim-3 might act as a potent regulator of antiviral T-cell responses in HBV infection. Cellular ＆ Molecular Immunology.     

Hepatitis B Virus Down-Regulates Expressions of MHC Class Ⅰ Molecules on Hepatoplastoma Cell Line

Chronic HBV infection is associated with a 100-fold high risk of developing hepatocellular carcinoma. Tumor recognition is of the most importance during the immune surveillance process that prevents cancer development in humans. In the present study, the expressions of MHC class Ⅰ molecules on hepatoplastoma cell line HepG2.2.15 were investigated to indicate the possible effects of HBV on the immune recognition during HBV-associated hepatocellular carcinoma. It was found that the expressions of MHC class Ⅰ molecules HLA-ABC, HLA-E and MICA were much lower in HepG2.2.15 cells compared with HepG2 cells. The expressing HBV in human hepatoplastoma cell line significantly down-regulated the expressions of MHC class Ⅰ molecules. Additionally, it was observed that in murine chronic HBsAg carriers the expression of classical MHC-I molecule on hepatocytes was down-regulated. These results demonstrated that HBV might affect the immune recognition during HBV- associated hepatocellular carcinoma such as the recognition of CD8^＋ T, NK-CTL and NK cells and prevent the immune surveillance against tumors. However, the effects of HBV down-regulation of MHC class I molecules on the target cells in vivo should be further studied. Cellular ＆ Molecular Immunology.     

HBV携带者血清病毒标志物和HBV DNA与肝组织中HBVcccDNA相关性的研究

Objective To investigate the correlation of sera HBV DNA and serological makers with hepatic tissue HBVcccDNA in chronic HBV carriers. Methods Real time fluorescence quantitative polymerase chain reaction (RT-PCR) were used to detect HBV covalently closed circular DNA (cccDNA) and total intrahepatic HBV DNA from 30 needle-biopsy specimens as well as HBV DNA in sera in chronic HBV carriers. Quantification of the HBsAg, HBeAg in sera were quantified using Chemiluminescence immunoassay. Results HBVcccDNA can be detected in chronic HBV carriers, which rang from 3.15 × 103 copies/mg to 1.06 × 107 copies/mg. There was a positive correlation between the cccDNA and HBVtDNA (r =0. 375, P < 0. 05 ), but there was no correlation between the cccDNA and sera HBV DNA (P =0. 174). There was a positive correlation between cccDNA and sera HBsAg quantification (r =0. 562, P <0. 001 ) but no correlation with sera HBeAg qantification ( r = 0. 152, P > 0. 05 ). Conclusion HBV cccDNA can be replicated stably in hepatic tissue in all chronic HBV carriers. HBV DNA in sera can not be indicated hepatic tissue cccDNA level. While HBsAg quantification in sera can be used as a marker of cccDNA quantification in hepatic tissue to some extent.     

HBV携带者血清病毒标志物和HBV DNA与肝组织中HBVcccDNA相关性的研究

Objective To investigate the correlation of sera HBV DNA and serological makers with hepatic tissue HBVcccDNA in chronic HBV carriers. Methods Real time fluorescence quantitative polymerase chain reaction (RT-PCR) were used to detect HBV covalently closed circular DNA (cccDNA) and total intrahepatic HBV DNA from 30 needle-biopsy specimens as well as HBV DNA in sera in chronic HBV carriers. Quantification of the HBsAg, HBeAg in sera were quantified using Chemiluminescence immunoassay. Results HBVcccDNA can be detected in chronic HBV carriers, which rang from 3.15 × 103 copies/mg to 1.06 × 107 copies/mg. There was a positive correlation between the cccDNA and HBVtDNA (r =0. 375, P < 0. 05 ), but there was no correlation between the cccDNA and sera HBV DNA (P =0. 174). There was a positive correlation between cccDNA and sera HBsAg quantification (r =0. 562, P <0. 001 ) but no correlation with sera HBeAg qantification ( r = 0. 152, P > 0. 05 ). Conclusion HBV cccDNA can be replicated stably in hepatic tissue in all chronic HBV carriers. HBV DNA in sera can not be indicated hepatic tissue cccDNA level. While HBsAg quantification in sera can be used as a marker of cccDNA quantification in hepatic tissue to some extent.     

Increased serum IL-10 in lupus patients promotes apoptosis of T cell subsets via the caspase 8 pathway initiated by Fas signaling

We sought to investigate the expression of Fas and FasL on T cell surface and caspase 8 involvement in T cell apoptosis promoted by serum IL-10 in systemic lupus erythematosus(SLE) patients.Cells and sera were obtained from 35 SLE patients.Apoptosis of T cells in patients with SLE was increased and associated with the SLE disease activity index(SLEDAI).Elevated expression of Fas and FasL on T cell surface contributed to increased apoptosis of T cells.Increased IL-10 in the sera of SLE patients was capable of inducing Fas and FasL expression on CD4~+T cell surface,promoting apoptosis of this cell subset.Decreased IL-10 serum levels and low expression of Fas were found in 5 patients of the first follow-up group after 2-month treatment.In another group with one-year treatment,the SLEDAI declined to inactive scores.Serum IL-10 was decreased significantly,and expression of Fas and FasL on T cells was also reduced.Declined apoptosis was predominant only in CD4~+T cell subset.When sera with high level of IL-10 were used to culture PBMCs from healthy controls,activated caspase 8 was elevated in CD3~+T,CD4~+T and CD8~+T cells.The study showed that serum IL-10 induced apoptosis of T cell subsets via the caspase8 pathway initiated by Fas signaling.Increased apoptosis of T cells contributes to autoantigen burden,which is pathogenic in the development of SLE.     

DNA vaccines: Mechanisms of action北大核心

BACKGROUND: In 1990, Wolff et al. reported that DNA was examined as a gene therapy tool, and emerged as a promising therapy after observations that simple injection of naked plasmid DNA and RNA led to profound transgene expression in vivo. DNA vaccines are recognized for harboring several distinguishing characteristics and advantages (including low cost, ease and rapidity of manufacturing, and stability) making them a method for addressing global health threats in the future. OBJECTIVE: To review the status and research progress of DNA vaccines in the view of mechanism of action: innate immune signaling from bacterial DNA, transfecting somatic cell by DNA vaccines, cross-presentation and cross-activation, transfecting antigen presenting cells by DNA vaccines, and apoptosis. METHODS: The first author retrieved the databases of PubMed and CNKI for the articles concerning DNA vaccines published between January 2000 and June 2017 using the keywords of “DNA vaccine, gene vaccine, plasmid DNA, cross-presentation, transfection, apoptosis” in English and Chinese, respectively. A total of 105 literatures were searched, and 47 eligible articles were included in accordance with the inclusion criteria. RESULTS AND CONCLUSION: The immunogenicity of DNA vaccines in humans has been limited by low expression levels of antigen, in comparison with the conventional protein vaccines in the past two decades. Studies on the mechanism of action of DNA vaccines in terms of antigen-presenting cell types able to cross-present DNA-encoded antigens, the activation of innate immune responses due to DNA itself and induction of cell apoptosis have suggested the opportunities to increase the immunogenicity of these vaccines. © 2018, Journal of Clinical Rehabilitative Tissue Engineering Research. All rights reserved.     

The immune response induced by hepatitis B virus principal antigens

Hepatitis B virus （HBV） infection occurs primarily in hepatocytes in the liver with release of infectious virions and non-infectious empty surface antigen particles into the bloodstream. HBV replication is non-cytopathic. Transient infections run a course of several months, and chronic infections are often life-long. Chronic infections can lead to liver failure with cirrhosis and hepatocellniar carcinoma. It is generally accepted that neutralizing anti-HBs antibodies plays a key role in recovery from HBV infection by containing the spread of infection in the infected host and facilitating the removal and destruction of viral particles. However, the immune response initiated by the T-cell response to viral antigens is also important for viral clearance and disease pathogenesis in HBV infection. The three structural forms of the viral proteins, the HBsAg, the particulate HBcAg, and the nonparticulate HBeAg, may preferentially elicit different Th cell subsets. The different IgG subclass profiles of anti-HBs, anti-HBc, and anti-HBe in different HBV infection status were revealed. Moreover, the different IgG subclass profiles in chronic carriers did not change with different ALT and AST levels and may reflect the difference between stimulating antigens, immune response, and the stages of viral disease and provide the basis for the use of vaccines and prophylactic treatments for individuals at high risk of human HBV infection. This review elucidates the detailed understanding of the immune responses induced during transient and persistent infection, and the development of immunotherapy and immunodiagnosis in patients with HBV infection, and possible means of reducing the liver damage.     

Intratumoral Expression of MIP-1β Induces Antitumor Responses in a Pre-Established Tumor Model through Chemoattracting T Cells and NK Cells

Direct intratumoral introduction of therapeutic or regulatory genes is a developing technology with potential application for cancer gene therapy. Macrophage inflammatory protein-1 beta (MIP-1β) is a chemokine which can chemoattract immune cells such as T cells. In the present study, murine colorectal adenocarcinoma CT26 cells were transfected with a recombinant adenovirus (AdhMIP-1β) carrying the human MIP-1β gene. 24h post-transfection, hMIP-1β levels reached approximately 980 pg/ml in supernatants of 10^6 hMIP-1β-transfected CT26 cells. Moreover, the supernatants exhibited chemotactic activity for CD8^ T cells, CD4^ T cells, NK cells and immature DCs. Intratumoral injection of AdhMIP-1β significantly inhibited tumor growth and prolonged the survival time of tumor-bearing mice. Intratumoral hMIP-1β gene transfer also induced powerful tumor-specific CTL responses in vivo. The therapeutic effects of hMIP-1β gene therapy were greatly reduced following in vivo depletion of both CD4^ and CD8~ T cells, but were unaffected by depletion of single T cell subsets. Immune cell depletion experiments also revealed that NK cells played an important role in hMIP-1β-induced antitumor responses. These results suggest that intratumoral expression of hMIP-1β has the potential effect to induce host antitumor immunity and may prove to be a useful form of cancer gene therapy.     

不同类型乙肝病毒感染者HBV抗原特异性T细胞免疫应答

目的 了解不同类型HBV感染人群T细胞对HBV抗原蛋白免疫应答的差别及特征.方法 76例研究对象分为四组,乙肝携带者组和既往乙肝感染患者组、急乙组、慢乙组,酶联免疫斑点法检测其外周血T细胞对HBV特异性抗原蛋白HBsAg、HBcAg和HBeAg免疫应答.结果 (1)携带者组对HBeAg的反应频率较高,而既往感染者对HBcAg和HBeAg的反应频率较高.急乙组和慢乙组对三种蛋白的反应频率无差别.(2)急乙组和慢乙组对HBsAg的反应频率明显高于既往感染组.急乙组、慢乙组和既往感染组对HBcAg的反应频率明显高于携带者组.对HBeAg的反应频率各组间无差别.(3)慢乙组对HBcAg的反应强度明显高于HBsAg.既往感染组患者对三种蛋白的反应强度依次为HBcAg＞ HBeAg＞ HBsAg.急乙组和携带者组对三种蛋白的反应强度无差别.(4)对HBsAg的反应强度从高到低依次为急乙组＞慢乙组＞携带者组和既往感染组.对HBcAg的反应强度是急乙组和慢乙组和既往感染组明显强于携带者组.对HBeAg的反应强度,急乙组高于慢乙组,其余各组间无差别.结论 急乙、慢乙和既往感染者对HBcAg的T细胞免疫反应为主,而携带者以HBeAg的T细胞免疫反应为主.     

Hepatitis B core antigen stimulates interleukin-10 secretion by both T cells and monocytes from peripheral blood of patients with chronic hepatitis B virus infection

In chronic hepatitis B virus (HBV) infection, immune responses to hepatitis B core antigen (HBcAg) are weak. Interleukin (IL)-10 is a potent immunosuppressive cytokine which we reported recently to be secreted in response to HBcAg by peripheral blood mononuclear cells (PBMCs) from patients with chronic HBV infection or healthy controls. Using an enzyme-linked immunospot assay, we compared the ability of HBcAg to stimulate IL-10 production by PBMC with that of lipopolysaccharide (LPS), phytohaemagglutinin-P and hepatitis C virus-derived antigens in 16 patients with chronic HBV infection and six healthy controls. Frequencies of IL-10 spot-forming cells (SFC) in response to HBcAg were comparable to those obtained with LPS in patients with chronic HBV infection. Frequencies of IL-10 SFC in response to HBcAg or to LPS were significantly higher in patients with chronic HBV infection than in healthy controls. IL-10 SFC in response to HBcAg consisted of 26-35% T cells, 62-70% monocytes and less than 1% B cells in patients with chronic HBV infection. Only monocytes contributed to IL-10 production in controls. Frequencies of HBcAg stimulated IL-10 SFC representing T cells and monocytes were significantly higher in patients with elevated serum alanine aminotransferase (ALT) and detectable HBV DNA than in patients with normal ALT and undetectable HBV DNA. The potent ability of HBcAg to stimulate IL-10 production by PBMC may contribute importantly to immune tolerance toward HBV.     

Application strategies of serum HBV DNA detection in HBV infection patients: A retrospective study of 5611 specimens

The detection of hepatitis B virus (HBV) DNA plays a critical role in determining the level of viral replication in HBV-infected patients. However, how to select appropriate HBV DNA detection method, low-sensitivity (ls) and hypersensitivity (hs) remains unclear. In this study, hepatitis B surface antigen (HBsAg), hepatitis B e-antigen (HBeAg), alanine transaminase (ALT), aspartate transaminase (AST), and hs HBV DNA titers in serum of 5611 cases with suspected HBV infection were reviewed. Besides, the dynamic changes of HBV DNA and HBsAg in 85 chronic hepatitis B (CHB) patients receiving peginterferon α (PegIFNα) or entecavir (ETV) were observed. The results showed the positive rate of HBV DNA was 32.8%, of which low viral load (20 to 500 IU/mL) accounted for 51.8%. In the 5611 cases, when the HBsAg was less than 1000 IU/mL, the proportion of low viral load was 76.3%. Moreover, in patients receiving antiviral treatment, when HBsAg was less than 2000 IU/mL (PegIFNα) or HBsAg was less than 3500 IU/mL (ETV), the proportion of patients with low viral load was 79.5% or 78.0%, respectively. We developed a strategy of serum HBV DNA detection in HBV-infected patients. When HBsAg was negative, HBV DNA detection should be unnecessary. When HBsAg was 0.05 to 1000 IU/mL, hs HBV DNA should be detected in patients with abnormal level of ALT, AST, or HBeAg. While HBsAg was greater than or equal to 1000 IU/mL, ls HBV DNA was recommended. Moreover, the cutoff value of HBsAg increased during antiviral therapy of CHB patients. In conclusion, hs HBV DNA is of great value in HBV-infected patients with low viral load. HBV DNA detection methods should be selected reasonably according to the levels of HBsAg, HBeAg, ALT, and AST.     

Serum M2BPGi level is a novel predictive biomarker for the responses to pegylated interferon‐α treatment in HBeAg‐positive chronic hepatitis B patients

Serum Mac‐2‐binding protein glycosylation isomer (M2BPGi) level was found to be a useful prognostic marker for hepatitis B e antigen (HBeAg)‐positive chronic hepatitis B (CHB) patients treated with nucleoside/nucleotide analogs (NUCs) therapy, and the aim of our study is to evaluate the clinical implementation of M2BPGi level in the prediction of antiviral responses to pegylated‐interferon‐α (PEG‐IFN‐α) treatment in HBeAg‐positive CHB patients. Ninety‐six CHB patients who received PEG‐IFN‐α treatment for at least 48 weeks were recruited. The serum M2BPGi, alanine aminotransferase (ALT), hepatitis B surface antigen (HBsAg), HBeAg, and HBV DNA levels at baseline, weeks 4, 12, and 24 after PEG‐IFN‐α treatment were determined and their associations with antiviral responses were evaluated and the virological response (VR) rate and serological response (SR) rate after 48 weeks of treatment were 65.6% and 35.4%, respectively. Baseline serum M2BPGi level was significantly different between VR and non‐VR (P = 0.002) or SR and non‐SR groups (P = 0.012). Multivariate analyses suggested that baseline serum M2BPGi level was independently associated with VR and SR of PEG‐IFN‐α treatment at week 48. The area under the ROC curve (AUC) of baseline M2BPGi was 0.682 in predicting VR, which was superior to HBsAg (AUC = 0.566) or HBV DNA (AUC = 0.567). The AUC of baseline M2BPGi in predicting SR was 0.655, which was also higher than that of HBsAg (AUC = 0.548) or HBV DNA (AUC = 0.583). These results suggested that baseline serum M2BPGi level was a novel predictor of VR and SR for PEG‐IFN‐α treatment in HBeAg‐positive CHB patients.     

Hepatitis B virus (HBV) antigen-pulsed monocyte-derived dendritic cells from HBV-associated hepatocellular carcinoma patients significantly enhance specific T cell responses in vitro

To investigate whether hepatitis B virus (HBV) antigen-pulsed monocyte-derived dendritic cells (MoDC) could mount a T cell response in hepatocellular carcinoma (HCC) patients associated with chronic HBV infection, peripheral blood mononuclear cells (PBMCs) from 36 HBV-associated HCC patients were induced into MoDC and pulsed with hepatitis B core antigen (HBcAg) and hepatitis B surface antigen (HBsAg), alone and in combination. Co-stimulatory molecules CD80, CD86 and CD40, as well as human leucocyte antigens D-related (HLA-DR) were found to express at the highest level on MoDC pulsed with HBcAg or HBsAg + HBcAg, at a median level on MoDC pulsed with HBcAg or HBsAg alone, and at the lowest level on non-antigen-pulsed MoDC. Interleukin (IL)-10 and IL-12 cytokines were released by antigen-pulsed MoDC at increased levels in the order: no-antigen < HBsAg < HBcAg < HBcAg + HBsAg. MoDC pulsed with HBcAg or HBsAg + HBcAg also had the strongest ability to stimulate autologous T cell proliferation and intracellular interferon (IFN)-gamma production. HBcAg- or HBsAg + HBcAg-pulsed MoDC could also induce HBV core peptide-specific CD8(+) T cell proliferation determined by tetramer staining. In addition, the antigen-pulsed MoDC were found to have a stronger capacity to produce IL-12 and induce T cell response in vitro for patients with higher alanine transaminase (ALT) levels than those with lower ALT levels, indicating that antigen pulse could substantially reverse the impaired function of MoDC in primary HCC patients with active chronic hepatitis B. In conclusion, HBV antigen-pulsed MoDC from HCC patients with chronic hepatitis B could induce HBV-specific T cell response in vitro.     

人树突状细胞体外经HBsAg刺激后的抗病毒作用

目的观察人树突状细胞(DCs)经表面抗原(HBsAg)刺激、体外诱导自身特异性T淋巴细胞增殖后,对2.2.15细胞中HBeAg和HBsAg的特异性免疫抑制作用。方法 用粒细胞-巨噬细胞集落刺激因子(GM-CSF)、白介素-4(IL-4)和肿瘤坏死因子(TNF-a)分化、诱导人外周血PBMC中的DCs,在DCs成熟前加入纯的HBsAg刺激,将成熟后的DCs体外与自身T淋巴细胞共培养,同时不加HBsAg刺激的DCs与T细胞共培养、T细胞加纯的HBsAg共培养以及单纯T细胞作对照,5 d后收集T细胞,分组加入2.2.15细胞培养液中,分别收集第1天、3天、5天和7天的培养上清液,检测其HBeAg和HBsAg的分泌情况。结果经抗原刺激后的DCs可以有效提呈病毒抗原,正常人与慢性乙肝患者负载抗原后的DCs刺激T淋巴细胞增殖的能力[cpm分别为(46 700±7 850)和(38 628±5 427)]明显高于未负载抗原的DCs[cpm分别为(40 450±4 645和33 924±4 498)]及对照组PBMC[cpm分别为(5 947±476)和(5 089±233)],P<0.01。负载抗原的DCs有强烈的免疫应答活性,并且其免疫刺激能力似乎与负载的抗原量成正比;经抗原刺激激活的T细胞可以有效地抑制HBeAg的表达,但对HBsAg未发现有明显的抑制作用。结论体外经HBsAg刺激后的DCs可有效地提呈病毒抗原,并可进一步激活T细胞产生,同时能显著地抑制2.2.15细胞上清     

肝组织病理及肝内抗原表达与聚乙二醇干扰素α-2a抗乙肝病毒疗效关系的研究

目的观察聚乙二醇干扰素α-2a（PegIFNα-2a）治疗慢性乙型肝炎的疗效,探讨肝脏病理改变和肝细胞内病毒抗原的表达类型与PegIFNα-2a抗病毒疗效的关系。方法选择HBeAg阳性慢性乙型肝炎68例,HBeAg阴性慢性乙型肝炎45例,通过肝组织病理检测,观察肝脏病理改变和肝细胞内病毒抗原的表达类型与PegIFNα-2a治疗后血清HBV DNA的阴转率、HBeAg转换率和完全应答率之间的关系,并随访48周,观察持续应答情况。结果 HBsAg的阴转在不同的炎症活动度间差异无统计学意义;在HBeAg阳性患者中,G1组的HBeAg转换率和完全应答率与G3组比较差异有统计学意义（P均〈0.05）;炎症活动度高的病例经治疗后48周的持续应答率高于炎症活动度低的病例（χ2=4.311,P〈0.05）;肝细胞内HBcAg浆型表达者HBeAg阴转、HBeAg转换率、HBV DNA的阴转率均高于HBcAg核型表达者（P均〈0.05）。结论肝组织病理改变和病毒抗原在肝细胞的表达类型可能成为PegIFNα-2a抗病毒疗效的潜在预测因素。炎症活动度高、肝细胞内HBcAg浆型表达者可能对PegIFNα-2a的治疗应答更好。     

Virus-specific lymphokine production differs quantitatively but not qualitatively in acute and chronic hepatitis B infection.

Cytokines that are secreted as a response to viral antigen not only have direct antiviral properties but also crucially influence immune reactions determining the outcome of infection. As an advantageous alternative to the study of cytokines present in the supernatants of antigen-specific T cell clones and lines, we have used ELISPOT assays to determine the number of interferon-gamma (IFN-gamma)- and IL4-producing cells generated by peripheral blood mononuclear cells from patients with acute hepatitis B (AHB) and chronic hepatitis B (CHB) infection in response to HBcAg in a short-term culture (48 h). In response to HBcAg IFN-gamma was predominantly produced. In contrast to the results obtained in acute hepatitis B, the typical lymphokine pattern in CHB was characterized by a weak or absent antigen-specific IFN-gamma production. A predominance of IL-4-producing cells was not observed in either AHB or CHB. A significant number of IFN-gamma-producing cells was usually detectable during phases of viral elimination and the quality of the lymphokine response seemed to be epitope independent. Comparison of the results obtained in proliferation assays and ELISPOT assays clearly shows that lymphokine production upon stimulation with viral protein is totally independent of T cell proliferation and more sensitively reflects antiviral reactivity.     

In vitro use of autologous dendritic cells improves detection of T cell responses to hepatitis B virus (HBV) antigens

T lymphocyte responses to hepatitis B virus (HBV) core antigen (HBcAg) are vigorous and easily detectable in vitro during recovery from acute hepatitis B but significantly weaker in patients with chronic HBV infection. In contrast, T cell responses to hepatitis B surface antigen (HBsAg) are almost undetectable during infection and even in a substantial fraction of subjects receiving vaccination with HBsAg. The aim of this study was to investigate whether the use of dendritic cells (DCs) in an in vitro assay could increase the detection of HBV‐specific T cells in these conditions. Autologous monocyte‐derived DCs, compared to direct HBsAg addition to the cultures, increased the stimulation of HBs‐ specific T cells. These were detected in 73% of healthy subjects who had recently received hepatitis B vaccine and in 43% of patients recovering from acute hepatitis B. Likewise, proliferation in response to DC‐presented HBcAg was detected in both CD4⁺ and CD8⁺ T cells from the majority of chronic hepatitis B patients. A longitudinal evaluation of HBc‐specific T cell responses during and after a 1‐year treatment with pegylated interferon (IFN)‐α showed that HBc‐specific CD4⁺ T cell responses had no correlation with sustained virus suppression whereas CD8⁺ T cell responses were more frequently detected in patients able to control HBV replication after therapy interruption. The use of autologous DCs as antigen‐presenting cells appears applicable to clinically relevant in vitro evaluation of T cell responses, particularly in those conditions characterized by low frequency of circulating antigen‐specific cells and suboptimal in vivo activation. J. Med. Virol. 81:332–339, 2009. © 2008 Wiley‐Liss, Inc.     

Long-term outcomes after nucleos(t)ide analogue discontinuation in HBeAg-positive chronic hepatitis B patients

Nucleos(t)ide analogue (NUC) resistance is an important clinical risk resulting from long-term therapy in chronic hepatitis B (CHB) management. Discontinuation of NUCs is a feasible strategy to reduce resistance. We aimed to observe the outcomes after NUC withdrawal in HBeAg-positive CHB patients. A total of 97 patients (11 patients with HBsAg loss and 86 patients with sustained HBeAg seroconversion) were enrolled. HBV DNA levels and alanine aminotransferase (ALT) levels were monitored regularly after discontinuation. Relapse was defined as HBV DNA levels >2000 IU/mL in at least two determinations more than 4 weeks apart. HBeAg seroconversion was achieved within 48 weeks (interquartile range (IQR), 24-72 weeks). The time on consolidation therapy was 96 weeks (IQR, 84-144 weeks). No relapses occurred for HBsAg loss patients. Evidence of relapse was observed in 9.3% of HBeAg seroconversion patients. All relapse cases occurred within 48 weeks after discontinuation. The time to relapse was 33 ± 15 weeks. Elevation of HBV DNA and ALT levels over baseline were only observed in 12.5% of relapse patients. There were no significant differences in baseline characteristics (sex, HBV genotype, age or ALT levels) or time on consolidation therapy between patients with relapse or sustained response. NUC discontinuation in HBeAg-positive CHB patients is feasible after achieving HBeAg seroconversion at a minimum of 24 weeks. There is further benefit to prolonging the time on consolidation therapy to reduce relapse. More than 48 weeks of sustained response is a predictive marker for long-term sustained response.