Antibodies to hepatitis B core antigen in hepatitis B surface antigen-positive and -negative chronic hepatitis.

Antibody to hepatitis B core antigen (anti-HBc) is a sensitive indicator of hepatitis B virus (HBV) infection. However, anti-HBc has been found in only a few patients with chronic hepatitis. Therefore, we tested for anti-HBc in 124 sera from 67 patients with histologically proven chronic hepatitis by the indirect fluorescent antibody technique. All patients, except for one with chronic hepatitis who was seropositive for hepatitis B surface antigen (HBs Ag), had anti-HBc that persisted throughout the follow-up period (three months to three years). Of 33 HBs Ag-seronegative patients, anti-HBc was detected in seven patients and persisted for six months to two years. These findings suggest that in this study 21% of patients with chronic hepatitis with undetectable amounts of HBs Ag in the serum had evidence of recent or continued HBV replication.     

Comparison of hepatitis B surface antigen and e antigen in predicting liver histology in hepatitis B e antigen-positive chronic hepatitis B patients

Background/Aim

The purpose of this study was to determine the relationship between hepatitis B e antigen (HBeAg) and hepatitis B surface antigen (HBsAg) levels and liver histology in HBeAg-positive patients with chronic hepatitis B virus (CHB) infection.

Methods

Serum HBsAg and HBeAg levels were analyzed, and liver biopsies were obtained from 203 HBeAg-positive CHB patients (62.6 % males; median age 31.3 years). The upper limit of normal (ULN) for ALT in this study was 30 and 19 U/L for males and females, respectively. Histologic assessment was based on Scheuer classification.

Results

ALT <2 × ULN, fibrosis stage <S2, and necro-inflammation grade <G2 were noted in 70 (34.5 %), 141 (69.5 %), and 105 (51.7 %) patients, respectively. Patients with significant histology (fibrosis stage ≥S2 and/or fibrosis stage of 1 plus inflammation grade ≥2) had significantly lower median HBsAg (13,998.4 and 42,186.5 IU/mL, respectively, p < 0.001) and HBeAg levels (540.5 and 1,206.8 S/CO, respectively, p < 0.001) than patients with insignificant histology. Among patients with ALT <2 × ULN, the area under the ROC curve for serum HBsAg and HBeAg levels was 0.76 and 0.70, respectively. Using a cutoff value of 17,558 IU/mL for HBsAg and 875.6 S/CO for HBeAg in patients with ALT <2×ULN, positive predictive values for insignificant histology were 87 and 86.8 %, respectively, whereas the negative predictive values were 50 and 47.1 %, respectively.

Conclusions

Among HBeAg-positive patients with ALT <2 × ULN, high-serum HBsAg and HBeAg levels can equally accurately predict insignificant histology.     

慢性乙型肝炎肝组织内HBsAg、HBcAg的表达及临床研究进展

一直以来临床将血清乙型肝炎e抗原(HBeAg)、乙肝病毒DNA(HBV DNA)阳性作为乙肝病毒复制的标志,随着肝穿活检及抗病毒治疗的研究进展,肝活检组织中乙肝表面抗原(HBsAg)和乙肝核心抗原(HBcAg)的表达模式与血清乙型肝炎病毒(HBV)DNA定量、肝组织炎症活动度分级及纤维化分期之间关系的临床研究日益增多,本文就HBsAg和HBcAg在肝组织的表达模式及临床研究进展综述如下.     

Significance of hepatitis B core antigen in the liver in patients with chronic hepatitis B and its relation to hepatitis B virus DNA

Liver biopsies from 52 patients with chronic hepatitis B were investigated for the presence and distribution of HBcAg and the results were compared with the status of hepatitis B virus deoxyribonucleic acid (HBV-DNA). The patients consisted of 37 men and 15 women, aged 16-55 years (mean = 34 years). Serum alanine aminotransferase (ALT) was elevated in 50 patients (range: 18-969 U/L; mean = 290 U/L). Serological testing showed HBsAg in all, HBeAg in 45 (87%), and HBV-DNA in 28 (54%). Liver biopsies demonstrated HBcAg in 35 (67%) patients. HBcAg was not only present in 31 of 45 (69%) patients who were seropositive for HBeAg, but also in four of seven (57%) with antibody to HBeAg (anti-HBe). In 28 of 35 (80%) patients with HBcAg in the liver, serum HBV-DNA was detected. However, no serum HBV-DNA was detected in 17 patients who had no detectable HBcAg in the liver. The distribution of HBcAg in the liver was rather cytoplasmic and nuclear than nuclear alone. Among 33 patients with cytoplasmic HBcAg in the liver, 15 (45%) had an evidence of acute exacerbation of hepatitis with marked ALT elevation (range: 168-894 U/L; mean = 385 U/L) and nine patients showed severe chronic active hepatitis and confluent necrosis, histologically. These results indicate that the presence of HBcAg in the liver correlates with the amount of circulating hepatitis B virus as quantified by serum level of HBV-DNA. The predominant cytoplasmic HBcAg in the liver may suggest the possibility of multiple episodes of acute exacerbation and more severe ongoing hepatitis during the clinical course.     

Therapeutic potential of a combined hepatitis B virus surface and core antigen vaccine in patients with chronic hepatitis B

Purpose

The safety and clinical efficacy of a vaccine containing both hepatitis B surface antigen (HBsAg) and hepatitis B core antigen (HBcAg) (HBsAg/HBcAg) were evaluated in patients with chronic hepatitis B (CHB).

Methods

Eighteen patients with CHB were administered a vaccine containing 100 μg of HBsAg and 100 μg of HBcAg. The vaccine was administered ten times at 2-weekly intervals, the first five times via the nasal route only and the subsequent five times via both nasal and subcutaneous routes. The safety and efficacy of this therapeutic approach were assessed by periodic assessment of the patients’ general condition, viral kinetics, and biochemical parameters during treatment and 24 and 48 weeks after therapy. The production of cytokines by peripheral blood mononuclear cells (PBMC) and antigen-pulsed dendritic cells (DC) was evaluated to assess the immunomodulatory effects of the HBsAg/HBcAg vaccine in CHB patients.

Results

The HBsAg/HBcAg vaccine was safe in all patients. No flare of HBV DNA or alanine aminotransferase (ALT) was recorded in any patient. Sustained HBV DNA negativity and persistently normalized ALT were detected in 9 (50 %) and 18 (100 %) patients with CHB, respectively. PBMC and HBsAg/HBcAg-pulsed DCs from HBsAg/HBcAg-vaccinated CHB patients produced significantly higher levels of various cytokines [interleukin 1β (IL-1β), IL-6, IL-8, IL-12, and tumor necrosis factor α (TNF-α)] than those from control unvaccinated CHB patients (p < 0.05) after stimulation with HBsAg/HBcAg in vitro.

Conclusion

HBsAg/HBcAg vaccine seems a safe and efficient therapeutic approach for patients with CHB.     

Complement fixing hepatitis B core antigen immune complexes in the liver of patients with HBs antigen positive chronic disease.

One hundred and fifty-two biopsies from serologically HBsAg positive and negative patients with liver disease were studied in immunofluorescence: for the presence of the surface (HBs) and the core (HBc) antigenic determinants foeterminants of the hepatitis B virus, of immunoglobulins and complement (C) deposits, and for the capacity to fix human C. Circumstantial evidence is presented suggesting that HBc immune-complexes are a relevant feature in the establishment and progression of chronic HBSAg liver disease. C fixation by liver cells was shown in all HBC positive patients with chronic hepatitis; an active form was present in every case, except two with a persistent hepatitis, an inverse ratio of HBc to C binding fluorescence being noted between active chronic hepatitis and cirrhotic patients. HBc without C fixation was observed in only three patients in the incubation phase of infectious hepatitis. IgG deposits were often found in HBc containing, C fixing nuclei. No C binding or IgG deposits were observed in acute self-limited type B hepatitis, in serologically positive patients with normal liver or minimal histological lesions, with and without HBs cytoplasmic fluorescence in their biopsy, or in serologically negative individuals.     

HBV感染者肝细胞凋亡与乙型肝炎表面抗原双染色研究

目的探讨乙型肝炎肝细胞凋亡与病毒感染的关系。方法应用原位末端标记技术和免疫组化方法对一组乙肝患者肝细胞凋亡和乙肝表面抗原（ＨＢｓＡｇ）表达状况同时进行了检测。结果受检的８例乙型肝炎患者肝组织中均可检出凋亡细胞，阳性信号位于细胞核以及肝窦内之凋亡小体中；阳性细胞在肝细胞索多散在分布，在肝细胞坏死灶中亦可检出凋亡细胞；凋亡多发生于病毒抗原弱表达细胞，而大多数抗原阳性细胞未检测到凋亡信号，此外，少数凋亡细胞ＨＢｓＡｇ呈阴性。结论乙型肝炎患者感染和未感染病毒的肝细胞均可发生凋亡，但凋亡只在少数细胞发生。     

Intrahepatic distribution of hepatitis B surface and core antigens in chronic hepatitis B virus infection. Hepatocyte with cytoplasmic/membranous hepatitis B core antigen as a possible target for immune hepatocytolysis

The intrahepatic distribution of hepatitis B core antigen (HBcAg) and surface antigen (HBsAg) was studied in 32 patients with chronic type B hepatitis, and the results were correlated with the status of hepatitis B e antigen/antibody (HBeAg/anti-HBe) and with the histologic activity of the patients. In HBeAg-positive patients with minor hepatitis activity, HBcAg was distributed mainly in the nuclei, whereas HBsAg was diffusely located on the plasma membrane as well as focally in the cytoplasm. In HBeAg-positive patients with chronic active liver disease, although the distribution pattern of HBsAg in liver remained unchanged, the expression of nuclear HBcAg decreased significantly with concomitant increase in cytoplasmic/membranous HBcAg expression. In HBsAg carriers who were anti-HBe positive, HBcAg was undetectable anywhere, whereas HBsAg could be seen only in the cytoplasm. These results suggest that membranous expression of HBsAg relates closely to active viral replication but is probably not responsible for the observed histologic activities. In contrast, cytoplasmic/membranous expression of HBcAg correlates with high degree of liver inflammatory activity. It is therefore suggested that hepatocytes with cytoplasmic/membranous HBcAg expression might be the target cells for immune hepatocytolysis.     

Geographical distribution of the subtype of hepatitis B surface antigen in Chinese.

Using the much more sensitive hemagglutination inhibition assay for subtyping of hepatitis B surface antigen (HBsAg), we examined the determinants a, d, w and r in 192 from 228 HBsAg positive adults who had been found after screening with reversed passive hemagglutination method. Sixty-four subtypable cases were asymptomatic carriers and the remaining 128 were liver disease patients. Among them there was no significant difference of the subtypes, invariably with adw as the main subtype. Geographical difference was evident: adr was the main subtype (78 per cent) among the northern Chinese; while adw was dominant (76 per cent) among the southern Chinese with the Yangtze River as a boundary. Eight of the 18 adr-subtyped northern Chinese were born and live in Taiwan where 91 per cent of HBsAg positive Taiwanese were adw-subtyped. This was an indirect evidence that intra-familial spreading from parents played an important role in hepatitis B virus infection.     

乙型肝炎病毒核心抗原在慢性乙型肝炎病毒感染者肝细胞内的分布及临床意义

目的 探讨HBcAg在慢性HBV感染者肝细胞内的分布情况及其临床意义.方法 对41例慢性HBV感染者进行肝组织穿刺,用免疫荧光组织化学技术在激光共聚焦显微镜下观察肝细胞内HBcAg的分布情况.计量资料采用Kruskal Wallis检验,计数资料采用卡方检验.结果 41例慢性HBV感染者中,36例肝细胞内HBcAg阳性表达,阳性率为87.8%,其中23例肝功能中度异常,10例肝功能轻度异常,3例肝功能正常.HBcAg有膜、胞质与核三种形式表达.在23例肝功能中度异常者中,6例呈明显膜型表达,无明显胞质型及核型,17例以胞质型与膜型混合表达,未发现核型表达.在10例肝功能轻度异常者中,以单纯胞质型为主,未见膜型与核型表达.在3例肝功能正常者中,以胞质型表达与少许核型表达为主,未见膜型表达.HBcAg表达类型与肝功能之间差异有统计学意义(χ2=10.60,P＜0.01).结论 慢性HBV感染者肝组织损伤与HBcAg的表达有直接联系,提示膜型表达的HBcAg是肝脏免疫损伤过程中的靶抗原.

Abstract：

Objective To explore the distribution and clinical significance of hepatitis B core antigen( HBcAg) in the hepatocytes of chronic hepatitis B virus infected patients. Methods Paraffin sections were made from 41 liver biopsy samples obtained from chronic hepatitis B virus infected patients. The immuno-fluorescence confocal technique was utilized to analyze the expression level and localization of HBcAg in hepatocytes. The data were analyzed by using Kruskal Wallis test and chisquare test. Results HBcAg expression were detected in 36 (87. 8%) patients, among whom 23 cases had moderate abnormal liver function, 10 with mild abnormal liver function and 3 with normal liver function. Among the cases with moderate abnormal liver function, 6 cases showed the simple membrane-type HBcAg expression, 17 cases showed mixed cytosolic-type and membrane-type HBcAg expression without the nuclear-type expression. Twelve cases with mild abnormal liver function only showed simple cytosolic-type HBcAg expression without membrane-type or nuclear-type expression. In the three patients with normal liver function, HBcAg was expressed in cytoplasm and nuclear but not on membrane. The correlation between HBcAg expression pattern and liver function was statistically significant (χ2 =10. 60, P＜0.01). Conclusion HBcAg expression is directly correlated with liver injury in chronic hepatitis B virus infected patients, which indicates that membrane expressed HBcAg is the target antigen during the immuno-attack of liver.     

Antibody to hepatitis B core antigen in patients with hepatocellular carcinoma.

Hepatitis B surface antigen (HBsAg), anti-HBs, and anti-HB core (HBc) were measured in 124 patients with hepatocellular carcinoma (HCC) in comparison with 299 control subjects of comparable ages, and in 48 cases of chronic hepatitis and 52 cases of hepatic cirrhosis. It was found that 72.6% of the HCC patients were positive for anti-HBc, and 80.6% were positive for at least one test, whereas in the control, anti-HBc was positive in 30.1% and 34.1% were positive for at least one test, the differences between the two groups being significant (P less than 0.01). The frequencies of positive tests for HBsAg and anti-HBc were the highest in HCC followed in decreasing order by cirrhosis, chronic hepatitis and the control group. A possible role of HB virus infection in hepatocellular carcinoma is discussed in relation to other factors.     

Detection of e antigen and antibody: correlations with hapatitis B surface and hepatitis B core antigens, liver disease, and outcome in hepatitis B infections.

Testing for e antigen and antibody (anti-e) was performed by immunodiffusion and counterelectrophoresis in patients with polyarteritis nodosa fulminant hepatitis, and chronic active hepatitis (CAH), in 59 asymptomatic carriers of hepatitis B surface antigen (HBsAg) who underwent liver biopsy, and in 150 carriers followed with sequential SGPT determinations. Counterelectrophoresis was more sensitive that immunodiffusion. Neither e antigen nor anti-e was found in the absence of HBsAg. Among HCsAg-positive patients with polyarteritis nodosa and CAH, e antigen was found in 16 of 18 and 13 of 22, respectively. It was not found in any of 43 patients with fulminant hepatitis, of whom 24 were HBsAg-positive. The e antigen was detected in none of 13 biopsied carriers with normal histology, 4 of 28 with nonspecific changes of 11 of 18 with CAH or chronic persistent hepatitis. Conversely, anti-e was present in 9 of 13 with normal biopsy, 7 of 28 with nonspecific changes, and none of 18 with CAH or chronic persistent hepatitis. The e antigen was found more commonly in nonbiopsied carriers with elevated SGPT, and anti-e in those with normal SGPT. Six carriers whose antigenemia terminated spontaneously had anti-e. The presence of e antigen correlated with a high titer of HBsAg, and with immunofluorescent detection of hepatitis B core antigen in the nuclei of hepatocytes. Conversely, anti-e was associated with significantly lower titers of serum HBsAg (P less than 0.001) and lack of detectable hepatitis B core antigen in the liver.     

Progressive and sufficient decrease of hepatitis B core antibody can predict the disappearance of hepatitis B virus DNA in Japanese patients with hepatitis B surface antigen clearance

The aim of this study was to elucidate the relationships among serum levels of hepatitis B virus (HBV) DNA, periods after hepatitis B surface (HBs) antigen clearance, and the titer of hepatitis B core (HBc) antibody in 200-fold diluted serum. Twelve patients who had clearance of HBs antigen from serum were studied. Five patients had not received any treatment (group A), and seven had received prednisolone withdrawal therapy. The patients in groups A and B were followed up for 86 months and 108 months (median), respectively. Serum HBV was measured by the nested polymerase chain reaction method. In both groups, serum HBV tended to become gradually undetectable after HBs antigen clearance. The positive rate of HBV in the sera 5 years or more after HBs antigen clearance was significantly lower than that in the sera at less than 5 years, both in group A (P = 0.004) and group B (P = 0.010). In both groups, the titer of HBs tended to decline every year after HBs antigen clearance. HBV was still detectable in the sera of some patients for a long period of time after they showed seroconversion to HBs antibody. The results suggest that detection of HBV was difficult in sera with an HBc titer of 30% or lower and at more than 5 years after HBs antigen clearance in both groups. It is important to note that HBV DNA rarely exists in the serum, even when HBs antigen and HBc are both negative. Received: November 22, 1999 / Accepted: April 28, 2000     

HBsAg阴性或抗-HBc阳性者肺癌术后辅助化学治疗中乙型肝炎病毒的再激活

目的 探讨HBsAg阴性或抗-HBc阳性者肺癌术后辅助化学治疗中HBV的再激活及其相关危险因素.方法 回顾性分析2003年1月到2011年12月接受辅助化学治疗的3280例肺癌术后患者,所有入组患者进行HBV血清学标志物和生物化学检测,并接受以顺铂为基础的辅助化学治疗方案.数据比较行x2检验.结果 367例HBsAg阴性或抗-HBc阳性肺癌术后患者中,14例(3.81％)进展为乙型肝炎.单因素分析表明,患者年龄≥70岁(x2=13.003,P=0.019)、肝脏CT检查为脂肪肝或早期肝硬化(x2=11.225,P=0.026)和使用糖皮质激素累计剂量超过150mg(x2=7.008,P=0.033)是辅助化学治疗中HBV再激活的相关因素;而性别、基础辅助化学治疗方案与HBV的再激活无关联.结论 肺癌术后的辅助化学治疗中,HBsAg阴性或抗-HBc阳性者有一定比例可以发生HBV的再激活.