总乙型肝炎核心抗原的抗体，乙肝病毒定量的非侵入性标记

StephenW

PLoS One. 2015 Jun 26;10(6):e0130209. doi: 10.1371/journal.pone.0130209.
Total Hepatitis B Core Antigen Antibody, a Quantitative Non-Invasive Marker of Hepatitis B Virus Induced Liver Disease.
Yuan Q1, Song LW1, Cavallone D2, Moriconi F2, Cherubini B2, Colombatto P2, Oliveri F2, Coco BA2, Ricco G2, Bonino F3, Shih JW1, Xia NS1, Brunetto MR2.
Author information

    1State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, National Institute of Diagnostics and Vaccine Development in Infectious Diseases, School of Public Health, Xiamen University, Xiamen, China.
    2Laboratory of Molecular Genetics and Pathology of Hepatitis Viruses, Hepatology Unit, Reference Center of the Tuscany Region for Chronic Liver Disease and Cancer, University Hospital of Pisa, Pisa, Italy.
    3Digestive and Liver Disease, General Medicine II Unit, University Hospital of Pisa, Pisa, Italy.

Abstract

Non invasive immunologic markers of virus-induced liver disease are unmet needs. We tested the clinical significance of quantitative total and IgM-anti-HBc in well characterized chronic-HBsAg-carriers. Sera (212) were obtained from 111 HBsAg-carriers followed-up for 52 months (28-216) during different phases of chronic-HBV-genotype-D-infection: 10 HBeAg-positive, 25 inactive-carriers (HBV-DNA≤2000IU/ml, ALT<30U/L), 66 HBeAg-negative-CHB-patients and 10 with HDV-super-infection. In 35 patients treated with Peg-IFN±nucleos(t)ide-analogues (NUCs) sera were obtained at baseline, end-of-therapy and week-24-off-therapy and in 22 treated with NUCs (for 60 months, 42-134m) at baseline and end-of-follow-up. HBsAg and IgM-anti-HBc were measured by Architect-assays (Abbott, USA); total-anti-HBc by double-antigen-sandwich-immune-assay (Wantai, China); HBV-DNA by COBAS-TaqMan (Roche, Germany). Total-anti-HBc were detectable in all sera with lower levels in HBsAg-carriers without CHB (immune-tolerant, inactive and HDV-superinfected, median 3.26, range 2.26-4.49 Log10 IU/ml) versus untreated-CHB (median 4.68, range 2.76-5.54 Log10 IU/ml), p<0.0001. IgM-anti-HBc positive using the chronic-hepatitis-cut-off" (0.130-S/CO) were positive in 102 of 212 sera (48.1%). Overall total-anti-HBc and IgM-anti-HBc correlated significantly (p<0.001, r=0.417). Total-anti-HBc declined significantly in CHB patients with response to Peg-IFN (p<0.001) and in NUC-treated patients (p<0.001); the lowest levels (median 2.68, range 2.12-3.08 Log10 IU/ml) were found in long-term responders who cleared HBsAg subsequently. During spontaneous and therapy-induced fluctuations of CHB (remissions and reactivations) total- and IgM-anti-HBc correlated with ALT (p<0.001, r=0.351 and p=0.008, r=0.185 respectively). Total-anti-HBc qualifies as a useful marker of HBV-induced-liver-disease that might help to discriminate major phases of chronic HBV infection and to predict sustained response to antivirals.

StephenW

公共科学图书馆之一。 2015年6月26日; 10（6）：e0130209。 DOI：10.1371 / journal.pone.0130209。
总乙型肝炎核心抗原的抗体，乙肝病毒定量的非侵入性标记性肝病。
袁Q1，宋LW1，Cavallone D2，Moriconi F2，凯鲁比尼B2，Colombatto P2，F2奥利韦里，可可BA2，Ricco的G2，博尼诺F3，施JW1，夏NS1，MR2布鲁涅托。
作者信息

    分子疫苗学和分子诊断，诊断研究所和疫苗开发的传染病国家重点实验室，公共卫生，厦门大学，厦门，中国的学校。
    2Laboratory分子遗传学和肝炎病毒的病理，肝病股，托斯卡纳地区的参考中心慢性肝病，癌症，比萨大学医院，比萨，意大利。
    3Digestive和肝脏疾病，一般医药II单位，比萨大学医院，比萨，意大利。

抽象

病毒性肝病非侵入性免疫标记是未满足的需求。我们测试的总数量和IgM，抗HBc在充分表征慢性乙肝表面抗原携带者的临床意义。色拉寺（212）从111例HBsAg携带者获得随访52个月（28-216）在慢性乙肝病毒基因型-D-感染不同阶段：10 HBeAg阳性，25不活动载波（HBV-DNA≤ 2000IU /毫升，ALT <30U / L），66 HBeAg阴性-CHB-患者和10与HDV-超感染。在35例PEG-IFN治疗±核苷（酸）IDE-类似物（NUCs）血清在基线获得，周-24-OFF-治疗NUCs（60个月，42处理结束治疗并在22 -134m）在基线和结束的后续行动。 HBsAg和IgM抗体抗-HBc由建筑师测定法（雅培，美国）测量;总-抗HBc双抗原夹心免疫测定法（万泰，中国）; HBV-DNA通过COBAS-的TaqMan（罗氏，德国）。总-抗-HBc均检测到的所有与血清HBsAg阳性携带者较低的水平而不CHB（免疫耐受，不活动，HDV-superinfected，中位数3.26，范围2.26-4.49日志10国际单位/毫升）与未处理-CHB（中位数4.68，范围2.76-5.54 LOG10国际单位/毫升）中，p <0.0001。 IgM抗体抗-HBc阳性使用慢性肝炎切客“（0.130-S / CO）呈阳性的212血清（48.1％），102。合计总额，抗-HBc和IgM，抗HBc显著相关（ P <0.001，R = 0.417）总-抗HBc显著慢性乙型肝炎患者的回应PEG-IFN（P <0.001）和NUC治疗的患者（P <0.001）有所下降;最低水平（中位数2.68，范围2.12-3.08日志10国际单位/毫升）被发现在谁清除乙肝表面抗原随后，在慢性乙型肝炎的（缓解和重新激活）total-和IgM抗-HBc阳性与ALT（P <0.001相关自发和治疗引起波动的长期反应， R = 0.351和p = 0.008，R = 0.185分别），总抗-HBc阳性资格作为乙肝病毒诱导的肝疾病的一个有用的指标，可能有助于区分慢性HBV感染主要阶段，并预测到抗病毒药物持续应答。

StephenW

Discussion

The dynamic quantification range of the new total-anti-HBc assay allows to distinguish chronic-HBV-infection associated with HBV-induced liver disease, namely chronic-hepatitis-B (CHB), from chronic HBV infection without HBV-induced liver damage, namely non-inflammatory HBeAg-positive and inactive HBeAg-negative phases, independently from HBV-genotypes. Our validation study confirms in well characterized HBsAg-carriers infected with genotype-D- HBV the results obtained in Asian patients infected with genotype B and C [21]; the highest levels of total-anti-HBc were detected in CHB, both HBeAg-positive and HBeAg-negative and the lowest levels in inactive-carriers (Fig 1). Very low levels of the antibody were reported in the non-inflammatory, HBeAg-positive immune-tolerant-phase in genotype-B and-C infections [21–23]. Consistently in our HBeAg-positive-carriers total-anti-HBc levels comparable to those of inactive-carriers were found only in an immune-tolerant carrier, whereas the remaining CHB-patients showed high antibody levels. Total-anti-HBc declined during antiviral and correlated with response to Peg-IFN±NUC at EOT in both REL and SVR and at EOF in SVR during NUC treatment (Figs 1 and 3). In SVR-patients total-anti-HBc levels declined reaching at EOF values comparable to inactive-carriers. Interestingly in NUC-treated patients, the lower total-anti-HBc serum levels corresponded to the longer follow-up in disease remission. Antibody levels were significantly lower in NUC-treated patients) tested about 60 months after starting therapy as compared with patients with Peg-IFN SVR tested 24-weeks after EOT (p = 0.002). The distribution pattern of IgM-anti-HBc in the same subgroups mimics that of total-anti-HBc, but unfortunately the dynamic quantification range of the IgM-assay hampers the diagnostic accuracy since half of values fall below the analytical specificity cut-off of the assay. Comparing the kinetics of the different HBV-markers according to response to Peg-IFN we found (Fig 3) that the decline of total-anti-HBc during therapy parallels that of HBV-DNA and this may be useful in clinical practice to confirm the effectiveness of Peg-IFN antiviral activity. However, during Peg-IFN-treatment total-anti-HBc is unable to distinguish REL from SVR who are instead identified better by HBsAg-kinetics as reported previously [17–20]. During NUC therapy the kinetics of total anti-HBc differ from those of the other HBV markers and the total-anti-HBc decline provides an added value to HBV-DNA and HBsAg monitoring beckoning the remission of HBV-induced liver damage under effective antiviral therapy. In patients treated with Entecavir or Tenofovir viral load is consistently suppressed in the very early phase of treatment, whereas HBsAg declines very slowly over time [17–20]. Interestingly long-term NUC-treated patients with very low levels of total-anti-HBc experienced the HBsAg-clearance with anti-HBs-sero-conversion during follow-up. Thus, quantification of total-anti-HBc might help to predict HBsAg loss and future long-term prospective studies should address the issue in larger cohort of patients. Finally, the findings that total-anti-HBc kinetics correlate with ALT levels as well as IgM-anti-HBc in both naturally-occurring [25–27] and therapy-induced remission and reactivation phases [26,27] support the view that total-anti-HBc is a reliable marker of HBV-induced liver disease in chronic HBsAg-carriers. In chronic HBeAg positive hepatitis B baseline total-anti-HBc levels were shown to predict HBeAg to anti-HBe seroconversion in Asiatic HBeAg-positive-CHB patients, treated with Peg-IFN or NUC infected with HBV genotype B and C [24]. Our findings underline the potential of the assay also in the management of chronic HBeAg-negative-CHB of the Mediterranean Area infected with HBV genotype D. In addition our work suggests that quantification of total-anti-HBc may be helpful to distinguish the inactive HBsAg carrier from HBeAg-negative-CHB which is characterized by intervening phases of disease remission and reactivation [25–26], to identify patients with higher chance of HBsAg clearance and to provide a new tool to answer the unmet needs for treatment tailoring [27–30].

All available data support the view that q-anti-HBc is complementary to HBsAg quantification. HBsAg is a product of HBV replication, ccc-HBV-DNA transcription and viral mRNAs translation whereas total-anti-HBc is expression of the antiviral immune response against the HBV “core” antigen. Our data suggest that symmetry or asymmetry of the two markers may have important diagnostic implications: high levels of HBsAg associated with low levels of total-anti-HBc are diagnostic for the immune-tolerance or florid non-inflammatory phase of HBeAg-positive HBV-infection, while high levels of both markers identify chronic hepatitis B (either HBeAg-positive and HBeAg-negative). In cirrhotic patients with advanced HBeAg-negative-CHB total-anti-HBc levels are high because of persistent HBV-induced liver disease whereas HBsAg may decline because of HBsAg deletion mutants which hamper HBsAg secretion. In treated patients the decline of total-anti-HBc parallel HBsAg-kinetics in patients who respond to anti-viral treatment (sustained responders to therapy), but is asymmetric with the unchanged HBsAg levels in patients whose hepatitis B recurs after treatment discontinuation (Relaspers). In addition the decline of HBsAg during antiviral therapy is HBV genotype dependent whereas the decline of total-anti-HBc is HBV genotype independent. Thus the combined used of both markers can improve the management of the HBsAg carrier consistently.

In conclusion total-anti-HBc as measured by double-antigen-sandwich-immune-assay is a reliable non invasive marker of HBV-induced liver disease helpful to identify chronic-HBV-infection associated with HBV-induced liver disease. Larger prospective studies are needed address to address its significance particularly in the clinical management of NUC-treated patients.

StephenW

讨论

新的总抗-HBc阳性检测的动态范围的量化可以区分乙肝病毒性肝病，慢性就是 - 乙型肝炎（CHB），慢性HBV感染HBV没有引起的肝损伤相关的慢性乙肝病毒感染，即非炎症HBeAg阳性和非活动HBeAg阴性阶段，独立于HBV-基因型。我们的验证研究证实在感染基因型-D- HBV感染B基因型和亚洲患者得到的结果很好地表征乙肝表面抗原携带者C [21];在慢性乙型肝炎患者共检出抗-HBc阳性的最高水平，HBeAg阳性和HBeAg阴性和非活动载波（图1）的最低水平。抗体的非常低的水平被报告在非炎性，HBeAg阳性免疫耐受相基因型-B和-C的感染[21-23]。一贯在我们的HBeAg阳性携带者的总抗HBc抗体水平相媲美的非活动携带者被发现仅在免疫耐受的载体，而其余CHB-患者表现出较高的抗体水平。在抗病毒药物的总抗一HBc下降，与相关回应PEG-IFN±NUC在EOT两个REL和SVR和EOF在SVR在治疗NUC（图1和图3）。在SVR-患者总抗HBc抗体水平下降，在到达EOF值可比非活动携带者。有趣的是在NUC治疗的患者中，较低的总抗-HBc阳性血清水平对应于更长期的随访在疾病缓解。抗体水平显著降低NUC治疗的患者）开始治疗，与患者用PEG-IFN SVR相比EOT（P测试后24周= 0.002）测试后约60个月。 IgM抗体抗-HBc在同一分组模仿分布格局的总抗-HBc阳性的，但不幸的是IgM检测的动态范围量化妨碍了诊断的准确性，因为半值低于分析特异性切断的化验。根据响应于聚乙二醇干扰素比较的不同的HBV-标志物的动力学，我们发现（图3），该总-抗HBc的治疗期间的下降平行即HBV-DNA的，这可能是在临床实践中，以确认有用PEG-IFN的抗病毒活性的功效。然而，在PEG-IFN治疗总-抗-HBc是无法从通过SVR的HBsAg-动力学如以前[17-20]报道谁是确定的，而不是更好地区分REL。在NUC治疗的总抗HBc的动力学不同于其他HBV标志物和总的抗-HBc阳性下降提供了一个附加价值，HBV-DNA与HBsAg监测招手下有效的抗病毒治疗乙肝引起的肝损害的不同缓解。与恩替卡韦或替诺福韦病毒载量治疗的患者始终压制在治疗的初期阶段，而HBsAg的下降速度非常慢随着时间的推移[17-20]。有趣的是长期NUC治疗的患者总-抗HBc水平很低经历的HBsAg清除与抗-HBs，血清转换在随访期间。因此，总的抗-HBc阳性的量化可能有助于预测HBsAg消失和未来长期的前瞻性研究应该在更大的患者人群解决这一问题。最后，该研究结果，即总-抗HBc动力学关联与ALT水平以及IgM抗体抗HBc在天然存在的[25-27]和治疗诱导缓解和再活化阶段[26,27]支持这样的观点总的抗-HBc阳性是乙肝病毒性肝病慢性乙肝表面抗原携带者的可靠标志。在慢性HBeAg阳性乙肝基线总-抗HBc水平可以预测到的HBeAg抗HBe血清转换在亚洲HBeAg阳性，慢性乙型肝炎患者，PEG-IFN或NUC感染HBV基因型B和C [24]处理。我们的研究结果强调也在地中海地区的慢性HBeAg阴性-CHB的管理感染HBV基因型D。另外，我们的研究表明，总-抗HBc的定量可能有助于区分非活动性HBsAg测定的电位从HBeAg阴性，乙肝载体的特点是通过干预缓解病情和活化[25-26]的阶段，以确定患者的HBsAg清除的机会较高，并提供了新的工具来回答治疗剪裁[27-28的未满足的需求30]。

所有可用数据支持的观点，即Q-抗-HBc互补的HBsAg定量。乙肝表面抗原是乙型肝炎病毒复制，CCC-HBV-DNA的转录和病毒mRNA翻译而总-抗HBc是抗HBV的“核心”抗原的抗病毒免疫反应的表达的产物。我们的数据表明，对称的两个标记的或不对称可具有重要的诊断意义：高水平的HBsAg与总抗HBc抗体水平低有关是诊断为免疫耐受的HBeAg阳性HBV-或红润非炎症相感染，而高水平的标志物识别慢性乙型肝炎（无论是HBeAg阳性和HBeAg阴性）。肝硬化晚期患者HBeAg阴性，乙肝的总抗HBc抗体水平是因为HBV持续性肝病，而乙肝表面抗原可能下降，因为HBsAg的缺失突变体，妨碍HBsAg的分泌高。在治疗的患者共抗-HBc阳性平行的HBsAg动力学患者的下降是谁的抗病毒治疗（持续应答，以治疗）响应，但不对称与不变HBsAg水平的患者，其乙肝治疗停药（Relaspers后复发）。此外乙肝表面抗原的过程中抗病毒治疗的下降是HBV基因型依赖性，而总的抗-HBc阳性的下降是HBV基因型无关。从而将合并使用两种标记的可提高HBsAg携带者的管理一致。

总之总-抗HBc为用双抗原夹心免疫测定法测量是乙型肝炎病毒性肝病一个可靠的非侵入性标记物有助于识别乙肝性肝病相关的慢性乙肝病毒感染。更大的前瞻性研究是需要地址来特别处理其意义NUC治疗的患者的临床管理。

StephenW

http://journals.plos.org/plosone ... ournal.pone.0130209

682256

核心抗原抗体总量，可以作为乙肝病毒引起疾病进程的非侵入型标记。

682256

也就是说，以后可以从核心抗原定量判断病情程度？