CpG ODN联合乙型肝炎疫苗与乙型肝炎疫苗免疫效果对比研究

目的探讨CpG ODN乙型肝炎疫苗与市售乙型肝炎疫苗对Balb/c小鼠的免疫作用效果。方法将乙肝疫苗、乙肝疫苗+100μgCpG ODN分别肌肉注射到4～6周龄,16～18g Balb/c小鼠体内,于第1次免疫后28d以同样剂量加强免疫1次。分别于第1次免疫后28、42、63 d收集小鼠血清,用ELISA方法检测抗HBs IgG抗体。结果实验组产生的抗HBs IgG与对照组HBsIgG相比P<0.05,具有显著性差异。28、42、63 d抗HBs IgG分别是对照疫苗的114倍、4.73倍、5.41倍以上。结论 CpG ODN能够显著增强小鼠对乙肝疫苗的免疫应答,在较短时间内显著的提高乙肝抗体的产生水平,免疫效果是乙肝疫苗的5倍以上。     

乙肝表面抗原突变体的表达及其初步应用

目的：构建和表达乙肝表面抗原（HBsAg）突变体用于HBsAg抗原性的深入研究。方法：利用定点突变技术构建HBsAg突变体，然后转化毕赤酵母GS115，菌落PCR、高浓度Zeocin抗性筛选鉴定转化子，表达产物经SDS—PAGE分析和Western blot分析，利用AxSYM HBsAg V2（Abbott）酶免试剂盒检测重组表面抗原的活性。结果：通过序列分析确定突变体构建成功，SDS—PAGE显示突变体能在毕赤酵母中有效表达，在Western blot试验中HBsAg突变体被特异性多克隆抗体识别，相对分子质量（Mr）约为38000，AxSYM试剂盒检测结果表明HBsAg突变体具有一定生物活性。结论：利用毕赤酵母表达系统高效表达出具有一定免疫反应性的HBsAg突变体。对于目前市售试剂盒的质量控制和临床应用有较高的实用价值。     

CpG-ODN对慢性乙型肝炎患者PBMC免疫刺激作用的观察

目的 观察CpG-ODN体外对慢性乙型肝炎患者PBMC的免疫刺激效应.方法 CpG-ODN体外刺激慢性乙肝患者外周血单个核细胞(PBMC),用ELISA检测培养液中IFN-α的水平;将不同比例的PBMC培养上清与HepG2.2.15细胞共孵育4和8 d后,用ELISA和荧光定量PCR法,分别检测培养上清液中HBsAg、HBeAg和HBV DNA的水平;MTT法观察活化的PBMC培养上清液对HepG2.2.15的杀伤作用.结果 CpG-ODN可有效诱导慢性乙肝患者PBMC分泌IFN-α,增强其活化的PBMC培养上清液对HepG2.2.15的杀伤作用;CpG-ODN本身虽不能直接抑制HBV的复制,但可通过其活化的PBMC的培养上清抑制HepG2.2.15细胞产生HBsAg、HBeAg和HBV DNA.结论 CpG-ODN可有效激活慢性乙肝患者的免疫细胞,发挥抗病毒效应.     

乙肝病毒表面抗原可疑阳性标本确认方法的建立

目的 建立基于酶联免疫吸附试验（ELISA）的乙肝病毒表面抗原（HBsAg）确认方法,避免错误诊断。方法收集ELISA检测HBsAb〉15COI的血清,取HBsAb阳性;HBsAb、HBeAb、HBcAb阳性;HBsAb、HBcAb阳性三种模式的血清各20份混合,与HBsAgELISA结果浓度〉20COI的血清中和以确认HBsAb的中和活性,制成确认试剂。确认试剂与30例HBsAg弱阳性阳性患者血清中和,HBsAgCOI下降率〉50％判定为阳性,其结果与罗氏确认试剂盒进行比对。结果确认试剂可以很好的中和HBsAg,对30例HBsAg弱阳性标本的判定结果为24例阳性,6例阴性,与罗氏确认试剂盒结果完全一致。结论成功建立HBsAg确认方法,适用于对HBsAg弱阳性样本进行确认,并且简单经济。     

Structural characterization of viral neutralizing monoclonal antibodies to hepatitis B surface antigen

In this study we describe the viral neutralizing activity of murine monoclonal antibodies (MAb) specific for hepatitis B surface antigen (HBsAg). This viral neutralizing activity was assessed in vitro by employing Hepatitis Delta Virus (HDV) and human hepatocytes as target cells. To further characterize these viral neutralizing antibodies we generated a panel of anti-idiotypic (anti-Id) reagents and serologically characterized these antibodies for epitope specificity, Id specificity, and Id heterogeneity. Direct binding and competitive inhibition solid phase enzyme immunoassay have demonstrated that two murine MAb specific for HBsAg (anti-HBs), designated A1.2 and A3.1, recognize similar or overlapping epitopes on HBsAg, while monoclonal anti-HBs, designated A2.1 recognizes a unique HBsAg epitope. Further, Id analysis using monoclonal and polyclonal anti-Id reagents have identified both a private and a cross-reactive Id, respectively, on the anti-HBs, A1.2 preparation. The source of the idiotypic cross-reactivity between A1.2 and A3.1 has been identified, using Western blot analysis, to conformational determinants expressed by the heavy (H) and light (L) chains of these monoclonal anti-HBs. Lastly, the intrastrain antibody repertoire induced following HBsAg immunization was found to be relatively restricted in heterogeneity by clonotype analysis using isoelectric focusing and affinity immunoblot analysis. Interspecies variability in the anti-HBs response was observed based on epitope recognition using purified anti-HBs from a variety of species.     

Long-term incidence and predictors of hepatitis B surface antigen loss after discontinuing nucleoside analogues in noncirrhotic chronic hepatitis B patients

ObjectiveTo investigate the long-term incidence and predictors for hepatitis B surface antigen (HBsAg) loss after nucleoside analogue therapy.MethodsThe study included 411 noncirrhotic chronic hepatitis B patients (148 hepatitis B e antigen (HBeAg)-positive and 263 HBeAg-negative patients) who were treated with lamivudine (n = 110) or entecavir (n = 301) with posttreatment follow-up of at least 12 months.ResultsIn HBeAg-positive patients, the 8-year cumulative rates of virologic relapse, clinical relapse and HBsAg loss were 55.6%, 47.7% and 19.6%, respectively. In HBeAg-negative patients, the rates were 69.3%, 58.9% and 33.1%, respectively. Cox regression analysis showed that hepatitis B virus genotype C and lower end-of-treatment HBsAg levels were independent predictors of HBsAg loss in HBeAg-positive and -negative patients. The 5-year HBsAg loss rate was 47.3% in HBeAg-positive patients with end-of-treatment HBsAg levels <300 IU/mL, while the 8-year HBsAg loss rate was 69.3% in HBeAg-negative patients with end-of-treatment HBsAg levels <200 IU/mL. Five patients experienced hepatitis flares with decompensation after stopping nucleoside analogue therapy, and one died after retreatment. Of the 48 patients who developed off-therapy HBsAg loss, two developed hepatocellular carcinoma.ConclusionsThe rate of HBsAg loss was relatively high and the rate of hepatic events was low in noncirrhotic patients who discontinued nucleoside analogue therapy.     

14株抗乙型肝炎表面抗原单克隆抗体与变异表面抗原的反应模式及应用

目的制备抗乙型肝炎表面抗原(hepatitis B surface antigen,HBsAg)的单克隆抗体,检测单克隆抗体与15种变异HBsAg的反应模式。用筛选出的单抗建立快速检测变异HBsAg的ELISA实验方法,并做初步评价。方法用中国乙型肝炎病毒感染者血清中分离的HBsAg免疫BALB/c小鼠,通过杂交瘤细胞融合技术制备抗-HBs单克隆抗体。检测不同单克隆抗体与野生及变异HBsAg的反应性。筛选出两种可以较好识别变异HBsAg的单克隆抗体McAb2和McAb3,建立两种抗体ELISA检测HBsAg的方法。结果制备了14株抗-HBs单抗。经过初筛,有4种可以较好识别包括G145R在内的大多数变异HBsAg。优化了McAb2和McAb3检测HBsAg的条件,检测HBsAg的灵敏度较好,检测变异HBsAg的能力优于2种现行国产HBsAg检测试剂盒。结论用本实验制备的单抗可以很好地识别包括G145R在内的大多数变异HBsAg。     

双歧杆菌完整肽聚糖对重组乙肝表面抗原的佐剂效应

目的 探讨双歧杆菌完整肽聚糖对重组乙肝表面抗原的吸附作用及其佐剂效应。方法 从双歧杆菌细胞壁提取完整肽聚糖，以不同方式与重组乙肝表面抗原结合，检测其对重组乙肝表面抗原的吸附作用；制备免疫原接种BALB／c小鼠，观察小鼠免疫后状态和生长情况，检测小鼠血清特异性抗体应答情况的变化。结果 在磷酸盐缓冲液中，完整肽聚糖对重组乙肝表面抗原的吸附呈剂量依赖性，作为佐剂免疫小鼠后无毒副反应出现，小鼠血清抗体阳性率和抗体滴度明显提高，IgC2a/IgG1增大。结论 双歧杆菌完整肽聚糖能够吸附重组乙肝表面抗原，增强机体相应的体液免疫应答和细胞免疫应答，具备良好的免疫佐剂效应。     

CpG-ODN联合HBsAg体外对慢性乙肝患者外周血树突状细胞STAT、SOCS通路的影响

目的 研究含非甲基化CpG基序的免疫刺激寡核苷酸（CpG-ODN）联合重组HBsAg对慢性乙肝（CHB）患者外周血树突状细胞（dendritic cell，DC）表面标志、功能及胞浆信号传导与转录激活因子（STAT）、细胞信号转导抑制因子（SOCS）表达的影响。方法由CHB患者和健康者外周血单核细胞诱导扩增DC，以CpG-ODN或联合HBsAg刺激DC，并与TNF-α比较，评价其对DC表面标志、分泌IL-12 p70及刺激同种T细胞增殖能力的影响；应用Western blot法检测DC胞浆STAT1、3、4、5、6以及SOCS1、3蛋白的表达。结果与PBS组比，TNF-α、CpG-ODN或联合HBsAg能明显提高CHB患者DC表面分子HLA-DR表达，IL-12 p70分泌增加，刺激同种T细胞增殖能力增强，CpG-ODN联合HBsAg还能明显提高CD1a的表达；CpG-ODN或联合HBsAg、TNF-α对DC胞浆STAT1、4、6和SOCS1、3的表达呈不同程度增强作用，对STAT3、5的表达无明显影响。结论CpG-ODN与TNF-α一样能促进CHB外周血DC分化、成熟；CpG-ODN或联合HBsAg能增强DC的特异性抗原提呈作用；其作用机制可能是通过调节DC细胞内STAT1、4、6以及SOCS1、3等信号蛋白的表达。     

Hepatitis B virus pre-genomic RNA and hepatitis B core-related antigen reductions at week 4 predict favourable hepatitis B surface antigen response upon long-term nucleos(t)ide analogue in chronic hepatitis B

Background/AimsWe investigated the dynamics of serum HBV pre-genomic RNA (pgRNA) and hepatitis B core-related antigen (HBcrAg) in patients receiving nucleos(t)ide analogues (NAs) and their predictability for favourable suppression of serum hepatitis B surface antigen (HBsAg).MethodsSerum viral biomarkers were measured at baseline, weeks 4, 12, 24, 36, and 48 of treatment. Patients were followed up thereafter and serum HBsAg level was measured at end of follow-up (EOFU). Favourable HBsAg response (FHR) was defined as ≤100 IU/mL or HBsAg seroclearance upon EOFU.ResultsTwenty-eight hepatitis B e antigen (HBeAg)-positive and 36 HBeAg-negative patients (median, 38.2 years old; 71.9% male) were recruited with median follow-up duration of 17.1 years (interquartile range, 12.8–18.2). For the entire cohort, 22/64 (34.4%) achieved FHR. For HBeAg-positive patients, serum HBV pgRNA decline at week 4 was significantly greater for patients with FHR compared to non-FHR (5.49 vs. 4.32 log copies/mL, respectively; P=0.016). The area under the receiver-operating-characteristic curve (AUROC) for week 4 HBV pgRNA reduction to predict FHR in HBeAg-positive patients was 0.825 (95% confidence interval [CI], 0.661–0.989). For HBeAg-negative patients, instead of increase in serum HBcrAg in non-FHR patients, FHR patients had median reduction in HBcrAg at week 4 (increment of 1.75 vs. reduction of 2.98 log U/mL; P=0.023). The AUROC for week 4 change of HBcrAg to predict FHR in HBeAg-negative patients was 0.789 (95% CI, 0.596–0.982).ConclusionsEarly on-treatment changes of serum HBV pgRNA and HBcrAg at 4 weeks predict HBsAg seroclearance or ≤100 IU/mL in NA-treated CHB patients upon long-term FU.     

重组乙肝表面抗原真核表达质粒pVAX-S2S的构建及DNA免疫

目的为探索更安全的乙型肝炎病毒(HBV)DNA疫苗,构建编码乙肝表面抗原中蛋白的卡那霉素抗性真核表达质粒,并观察其诱导BALB/c小鼠产生体液免疫应答情况.方法采用限制性内切酶从重组的真核表达质粒pcDNA-S2S中分离出乙肝表面抗原中蛋白(preS2+S)基因片段,将其亚克隆于pVAX1真核表达载体,酶切鉴定,按不同剂量一次性肌内注射免疫小鼠,ELISA法检测小鼠血清抗-HBs.结果酶切鉴定重组质粒pVAX-S2S为正向插入的阳性克隆.HBVDNA疫苗(pVAX-S2S)高(100μg/只)、中(50μg/只)、低(10μg/只)三组剂量一次性免疫健康BALB/c小鼠,均能在2 w诱导抗-HBs产生,抗体效价随时间延长而增长.血清抗体水平比较,高剂量组(97.83±38.78)mU/ml较中剂量组(45.13±21.12)mU/ml、低剂量组(19.74±11.92)mU/ml差异均具显著性(P＜0.05),以后的4,8 w高、中剂量组间差别缩小,但两者较低剂量组差异均具显著性(P＜0.05)和非常显著性(P＜0.01).结论卡那霉素抗性的重组质粒pVAX-S2S能有效诱导正常小鼠产生体液免疫应答.     

Association of concurrent hepatitis B surface antigen and antibody to hepatitis B surface antigen with hepatocellular carcinoma in chronic hepatitis B virus infection

Antibody to hepatitis B surface antigen (HBsAg) (anti‐HBs) can exist in patients with chronic hepatitis B virus (HBV) infection. To date, little is known about the association of concurrent HBsAg and anti‐HBs (concurrent HBsAg/ anti‐HBs) with hepatocellular carcinoma (HCC). The aim of this study was to investigate the clinical relevance of concurrent HBsAg/anti‐HBs with preS deletion mutations and HCC in chronic HBV infection. A total of 755 patients with chronic HBV infection were included consecutively at a tertiary center. Logistic regression analysis was used to identify risk factors for HCC, and serum HBV DNA was amplified, followed by direct sequencing to detect preS deletions. The prevalence of concurrent HBsAg/anti‐HBs was 6.4% (48/755) and all HBVs tested were genotype C. HCC occurred more frequently in the concurrent HBsAg/anti‐HBs group than in the HBsAg only group [22.9% (11/48) vs. 7.9% (56/707), P = 0.002]. In multivariate analyses, age >40 years [odds ratio (OR), 14.712; 95% confidence interval (CI), 4.365–49.579; P < 0.001], male gender (OR 2.431; 95% CI, 1.226–4.820; P = 0.011), decompensated cirrhosis (OR, 3.642; 95% CI, 1.788–7.421; P < 0.001) and concurrent HBsAg/anti‐HBs (OR, 4.336; 95% CI, 1.956–9.613; P < 0.001) were associated independently with HCC. In molecular analysis, preS deletion mutations were more frequent in the concurrent HBsAg/anti‐HBs and HCC groups than in the HBsAg without HCC group (42.3% and 32.5% vs. 11.3%; P = 0.002 and 0.012, respectively). In conclusion, concurrent HBsAg/anti‐HBs is associated with preS deletion mutations and may be one of the risk factors for HCC in chronic HBV infection with genotype C. J. Med. Virol. 81:1531–1538, 2009. © 2009 Wiley‐Liss, Inc.     

Detection of hepatitis B surface antigen (HBsAg) in peripheral blood mononuclear cells of patients with acute and chronic active hepatitis B

Seventy five patients with acute and chronic active hepatitis (CAH) were studied by indirect immunofluorescence with monoclonal antibodies for the presence of hepatitis B surface antigen (HBsAg) on peripheral blood mononuclear cells (PBMC). The viral surface antigen was detected in the PBMC of all the patients with hepatitis B virus (HBV)-induced CAH and in acute patients with more than 2 months of evolution. No HBsAg was detected in the samples obtained from 12 normal controls or from 14 non-A, non-B CAH patients. Analysis of PBMC subsets revealed that HBsAg was present in non-T cells; dual fluorescence studies showed HBsAg on surface Ig-positive lymphocytes. The binding of anti-HBs monoclonal antibodies was higher than that of a goat anti-HBs serum, and the highest reactivity was observed with an antibody against the pre-S(2)-region sequence. Both HBsAg and hepatitis B core antigen (HBcAg) were also detected in lysates of PBMC by dot blot analysis.     

Similar as well as distinct MHC class I-binding peptides are generated by exogenous and endogenous processing of hepatitis B virus surface antigen

Murine MHC class I-restricted cytotoxic T lymphocyte (CTL) responses can be primed by exogenous as well as endogenous hepatitis B surface antigen (HBsAg). Immunodominant CTL-defined epitopes of this viral envelope protein are the L^d -binding 12-mer S28 – 39 peptide IPQSLDSWWTSL in H-2 ^d mice, and the K^b -binding 8-mer S208 – 215 peptide ILSPFLPL in H-2^b mice. We tested if CTL recognizing these epitopes can be primed in vivo by HBsAg delivered as either an exogenous antigen (native HBsAg lipoprotein particles), or an endogenous antigen (plasmid DNA encoding HBsAg). Primed T cells were restimulated in vitro prior to the cytotoxicity assay with cells presenting the H-2 class I-binding epitopes generated by either exogenous or endogenous processing of HBsAg. The data indicate that the L^d -binding peptide S28 – 39 is generated during exogenous as well as endogenous processing of HBsAg. In contrast, the K^b -binding peptide S208 – 215 is generated during exogenous but not endogenous processing of HBsAg. Hence, some but not all MHC class I-binding, immunogenic peptides are generated during endogenous and exogenous processing of HBsAg but there also exists a repertoire of immunogenic peptides of viral origin that is only revealed after exogenous processing of viral proteins.     

Hepatitis B surface antigen levels at 6 months after treatment can predict the efficacy of lamivudine-adefovir combination therapy in patients with lamivudine-resistant chronic hepatitis B

Background/Aims

Quantitation of hepatitis B surface antigen (HBsAg) is an increasingly popular method to determine the treatment response in chronic hepatitis B (CHB) patients. The clinical value of HBsAg level measurement during rescue therapy for lamivudine (LMV)-resistant CHB patients have not been evaluated to date. Therefore, this study investigated the correlation between HBsAg level and treatment response in LMV-resistant CHB patients treated with adefovir (ADV) add-on therapy.

Methods

LMV-resistant CHB patients treated with LMV-ADV combination therapy for over 2 years were included. HBsAg levels were measured at 6 month intervals until 1 year, and annually thereafter. Treatment response was assessed by determining the virological response (VR, undetectable HBV DNA levels) during treatment.

Results

Fifty patients were included, of which 40 showed a VR. HBsAg levels were not different significantly at baseline (4.0 vs. 3.6 Log10 IU/mL, P=0.072). However, the HBsAg level decreased after 6 months of treatment in patients with a VR and became different significantly between the groups thereafter (3.9 vs. 3.3 at 6 months, P=0.002; 3.8 vs. 3.2 at 1 year, P=0.004; 3.9 vs. 3.2 at 2 years, P=0.008; 3.7 vs. 3.1 at 3 years, P =0.020).

Conclusions

The HBsAg level at 6 months after treatment can help predict treatment response.     

HBsAg和HBsAb双阳性乙肝患者血清中HBsAb确认方法的建立

目的 建立一种基于酶联免疫吸附实验（ ELISA）的HBsAb确认方法,验证HBsAg和HBsAb双阳性的乙肝患者血清中HBsAb阳性的真实性,剔除假阳性结果,避免错误诊断.方法收集60例电化学发光免疫分析法（ ECLIA）检测的HBsAg浓度在1000 COI以上的混合血清作为确认血清,将不同稀释度的确认血清与收集的HBsAb阳性混合血清中和,筛选并确定确认血清中HBsAg的最佳试验浓度.收集40例HBsAg和HBsAb双阳性的血清,与确认血清中和后采用ELISA检测COI的下降率,验证HBsAg和HBsAb双阳性标本中HBsAb阳性的真实性.结果 确认血清HBsAg浓度为2000 COI时对HBsAb的中和效果最好,ELISA确认法对40例HBsAg和HBsAb双阳性标本确认结果为37例真阳性和3例假阳性,与ECLIA法完全一致.结论 成功建立了HBsAg和HBsAb双阳性血清HBsAb的确认方法,该方法简单、准确且成本较低.     

Anti-hepatitis B surface antigen IgG1 subclass is predominant in individuals who have recovered from hepatitis B virus infection, chronic carriers and vaccinees

Patterns of each IgG-specific subclass for hepatitis B virus (HBV) core antigen (anti-HBc) are remarkably different among individuals with different infection status, i.e., completely recovered or chronic carrier. Each of the IgG-specific subclasses of HBV surface antigen (anti-HBs) was tested for ELISA sensitivity using four commercially available hepatitis B surface antigen (HBsAg) kits and one self-prepared plate. The specificity in 18 serum samples obtained from chronic HBV carriers, recovered individuals, vaccinees and non-infected individuals was investigated. Differences in absorbance values were obtained by comparing results from these different plates. Data on the absorbance values of anti-HBs IgG subclasses obtained indicated that one to four subjects had a false-negative or false-positive result using the four commercial plates. Only the self-prepared plate demonstrated 100% specificity and sensitivity for anti-HBs subclasses. Moreover, the results indicate that anti-HBs subclass IgG1 was predominant in cured patients, chronic carriers and vaccinees. The samples from both chronic carriers and vaccinees exhibited a significantly higher concentration of total IgG and IgG1 than samples in recovered individuals (P<0.05).     

Demonstration of hepatitis B surface antigen by orcein staining in paraffin sections of cirrhotic liver

Summary Blood and liver from 44 and 30 patients, died in cirrhosis and other diseases, respectively, were tested for the presence of HB_sAg. Blood samples obtained at autopsy and in seven cirrhosis cases also before death were tested for HB_sAg by counter-electrophoresis. Detection of HB_sAg in hepatocytes was carried out in paraffin sections by the modified orcein staining technique of Shikata et al. Ten of 14 HB_sAg seropositive and 2 of 30 HB_sAg seronegative cirrhotic patients had orcein positive hepatocytes, which were not found in any liver specimen from the 30 non-cirrhotic seronegative patients. The orcein positive substance localized in the cytoplasm of hepatocytes, less often it was also seen in a few Kupffer cells. The hepatocellular carcinoma cells present in part of the livers studied did not contain any orcein positive substance. Histological changes in the cirrhotic livers showed no morphological indication of the presence of HB_sAg, except on staining with orcein. The modified orcein staining technique is a simple, handy procedure for use in any routine pathological laboratory and has the additional advantage of detecting HB_sAg also in stored paraffin blocks.The authors are indebted to Dr. K. Vedrödi, Mrs. G. Molitorisz, Miss É. Gazsik and Miss Zs. Csicskár for performing the serological examinations for HB_sAg and anti-HB_s.     

Peroxidase-anti-peroxidase detection of hepatitis B surface and core antigen in liver biopsy specimens from patients with chronic type B hepatitis

Liver biopsy specimens from 58 American patients with chronic type B hepatitis were investigated for the presence and distribution of the hepatitis B core (HBcAg) and surface (HBsAg) antigens by peroxidase-anti-peroxidase techniques. HBsAg was detected in 43 (77%) and HBcAg in 52 (90%) patients. HBcAg was present in 50 of 51 (98%) patients with hepatitis B e antigen (HBeAg) but in only two of seven (29%) of patients with antibody to HBeAg (anti-HBe). There was no correlation between severity of hepatitis or height of aminotransferase activities and the amount of HBsAg or HBcAg in hepatocytes but there was a positive correlation between amount of HBcAg and height of HBV-DNA and DNA polymerase activity in serum. Follow-up liver biopsies, taken 1 to 3 yr later, were available from 39 patients. HBcAg remained detectable in 25 of 26 patients with persistence of HBeAg but disappeared in 12 patients who had lost HBeAg. In nine patients, HBcAg was cytoplasmic as well as nuclear in distribution. Seven of these patients had an intense lobular hepatitis with marked elevations in aminotransferase activities. These findings indicate that the amount of HBcAg in liver correlates with the amount of serum hepatitis B virus as quantified by serum levels of DNA polymerase and HBV-DNA. The amount of nuclear HBcAg does not correlate with the severity of the liver disease, but the presence of cytoplasmic HBcAg usually reflects an active and severe ongoing hepatitis.