Biomarcadores convencionales y emergentes en hepatitis B / Conventional and emerging biomarkers in hepatitis B

A nivel mundial, 300 millones de personas están infectadas por el virus de la hepatitis B (VHB). A pesar de que existe una vacuna que previene la infección y se dispone de tratamiento antiviral que suprime la replicación del virus, no hay cura aún. El principal problema que evita la recuperación total del paciente, incluso para aquel que recibe tratamiento, es la persistencia de dos formas del genoma viral en los hepatocitos: el ADN circular covalentemente cerrado (ADNccc), el cual se encuentra en forma de episoma y tiene la capacidad de replicarse, y las secuencias lineales subge-nómicas que se integran en el genoma humano, con potencial oncogénico. Hasta el momento se dispone de unos pocos biomarcadores para monitorear o predecir la progresión de la enfermedad y la respuesta al tratamiento. Estos biomarcadores se detectan durante la infección, y son la base para la monitorización de la enfermedad y hacer un diagnóstico de la fase clínica de la infección. Recientemente han surgido nuevos biomarcadores como el antígeno relacionado con el core del virus de la hepatitis B (HBcrAg) y la detección del ARN del VHB, que parecen correlacionarse con los niveles transcripcionales del ADNccc, además, durante el tratamiento parecen ayudar a predecir la respuesta y podrían identificar aquellos a quienes se les puede suspender la terapia sin riesgo de recaída. En esta revisión, se describe la utilidad de los principales biomarcadores convencionales en hepatitis B, y se abordan los dos biomarcadores emergentes más estudiados que prometen evaluar el curso de la infección, al igual que determinar la progresión de la enfermedad y la respuesta al tratamiento.

Globally, 300 million people are infected with hepatitis B virus (HBV). Although there is a vaccine that prevents infection and antiviral treatment that suppresses the replication of the virus, there is still no cure. The main problem that prevents the total recovery of the patient, even for those who recei-ve treatment, is the persistence of two forms of the viral genome in hepatocytes: covalently close circular DNA (cccDNA), which is in the form of an episome that has the ability to replicate, and linear subgenomic sequences that are integrated into the human genome, with oncogenic potential. Few biomarkers are currently available to monitor or predict disease progression and response to treatment. These biomarkers are detected during infection and are the basis for monitoring the di-sease and making a diagnosis of the clinical phase of the infection. New biomarkers have recently emerged, such as hepatitis B core-related antigen (HBcrAg) and HBV RNA detection, which seem to correlate with cccDNA transcriptional levels while during treatment seem to help predict response, and could identify those for whom therapy can be discontinued without risk of relapse. In this review, the usefulness of the main conventional biomarkers in hepatitis B is described, and the two most studied emerging biomarkers are mentioned, which promise to evaluate the course of the infection, as well as to determine disease progression and treatment response.

Clinical and pathological characteristics of chronic hepatitis B patients with normal alanine aminotransferase and not receiving antiviral therapy / 中华微生物学和免疫学杂志

Objective:To analyze the clinical and pathological characteristics of chronic hepatitis B (CHB) patients with normal alanine aminotransferase (ALT) and not receiving antiviral therapy.Methods:This study retrospectively included CHB patients diagnosed by liver biopsy at the Third Hospital of Hebei Medical University from January 2008 to December 2022. According to the HBV DNA and HBeAg status of "immune tolerance period and immune control period", these patients were divided into three groups: chronic HBV carrier group, inactive HBsAg carrier group and indeterminate group including the patients that did not meet the inclusion criteria of the above two groups. Kruskal-Wallis H test was used for comparison of continuous data between multiple groups. Mann-Whitney U test was used for comparison of continuous data and ordered categorical data between two groups. Chi-square test or Fisher′s exact test was used for comparison of categorical data between two groups. Results:A total of 284 CHB patients with normal ALT were enrolled. There were 64, 88 and 132 cases in the chronic HBV carrier group, inactive HBsAg carrier group and indeterminate group, respectively. Histopathological analysis revealed that there were 182 (64.08%) cases with pathological inflammation grade (G) and/or fibrosis stage (S)≥2, 155 (54.58%) with S≥2 and 120 (42.25%) with G≥2. The proportion of patients with G and/or S≥2 in the indeterminate group [70.45% (93/132)] was higher than that in the chronic HBV carrier group [48.44% (31/64)] and inactive HBsAg carrier group [65.91% (58/88)] (both P<0.05). Patient′s age and the ratio of patients with S≥2 in the chronic HBV carrier group [33 years old, 39.06% (25/64)] were smaller than those in the inactive HBsAg carrier group [39 years old, 56.82% (50/88)] and the indeterminate group [39 years old, 60.61% (80/132)] (all P<0.05). Patients in the inactive HBsAg carrier group (19 U/L) had lower ALT levels than those in the chronic HBV carrier group (26 U/L) and the indeterminate group (23 U/L) (both P<0.05). The proportion of patients with cytoplasmic/cytoplasmic nuclear-type HBcAg was higher in patients with G and/or S≥2 than in patients with G and S<2 [73.08% (57/78) vs 32.08% (17/53), P<0.05], and the proportion of patients with cytoplasmic/cytoplasmic nuclear-type HBcAg increased gradually with age. The proportion of patients with cytoplasmic/cytoplasmic nuclear-type HBcAg was higher in patients with G and/or S≥2 than in patients with G and S<2 in the chronic HBV carrier status and indeterminate groups [93.33% (28/30) vs 43.33%(13/30), P<0.05; 59.46% (22/37) vs 12.50% (2/16); both P<0.05]. There was a statistically significant difference in the incidence of significant liver injury between patients≤ 30 years old and >30 years old [52.7% (39/74) vs 68.1% (143/210), P<0.05]. Conclusions:Significant liver injury occurred in 64.08% (182/284) of CHB patients with normal ALT not receiving antiviral therapy, which required the attention of clinicians. Among CHB patients with normal ALT, the expression site of HBcAg in hepatocytes was related to the occurrence of significant liver injury and could be expected to serve as an important indicator for predicting the patient′s status and the necessity of antiviral treatment. CHB patients with positive HBV DNA who were older than 30 years required antiviral treatment, and CHB patients≤30 years with normal ALT and significant hepatic tissue damage also required antiviral treatment.

Mutations in C region of hepatitis B virus may contribute to occult hepatitis B virus infection / 实用医学杂志

Objective To investigate the role of mutations in C region that may contribute to occult hepatitis B virus infection.Methods C genes were amplified from two OBI blood donor samples respectively.Plasmids with mutations in C region of hepatitis B virus were constructed by overlapping PCR.HBsAg and HBeAg in Huh7 cells and in the serum of Balb/c mice were detected by CMLA.HBcAg in liver tissue was detected by immunohistochemistry,while HBV-RNA was tested by RT-PCR.Results Mutations in C region significantly reduced the expression level of HBeAg and HBcAg,but had no significant effect on HBsAg and HBV-RNA.Conclusion The mutations in C region affect the expression level of HBeAg and HBcAg,which may play an important role in the occurrence of OBI.

Correlations betweenintrahepatic HBsAg and HBcAg levels and serum HBsAg and HBV DNA levels in low-level HBsAg patients / 实用医学杂志

Objective To analyze the relationship between intrahepatic expression of HBsAg and HBcAg and the serum HBsAg and the load of HBV DNA in low-level HBsAg.To provide the basis for the diagnosis of low-level HBsAg.Methods A total of 63 patients with HBsAg lower than 1 400 IU/mL and higher than 0.05 IU/mL were enrolled in the study.All the 63 patients with low level HBsAg were collected for liver biopsy from January 2013 to December 2015.Serum HBsAg was detected by Roche cobas e601 automatic electrochemiluminescence immunoas-say system and supporting reagents.HBV DNA was detected using ROCHE automatic AmpliPrep/cobas TaqMan 48 system detector and supporting reagents.Results Among the 63 patients,the HBsAg in liver tissue was negative in 5 cases(7.94%),+in 47 cases(74.60%),++in 8 cases(12.70%),+++in 2 cases(3.17%),and++++was found in 1 case.The expression of HBcAg were negative in 30 cases(47.62%),+in 28 cases(44.44%),++in 4 cases(6.35%),+++in 1 case(1.59%),and++++in 0 cases.There was a significant positive correlation between HBsAg expression and serum HBsAg level by spearman and kendall correlation analysis(spearman analysis:r = 0.261,P=0.039;kendall analysis:r=0.217,P=0.036).HBcAg expression was significantly correlated with serum HBV DNA load with spearman and kendall correlation analysis(spearman analysis:r=0.305,P=0.015;kendall analysis:r=0.259,P=0.017).Conclusions The expression of HBsAg in liver tissue of in the patients with low level of HBsAg is positively correlated with the level of serum HBsAg,but not with the load of HBV DNA. There is significant positive correlation between HBcAg expression and serum HBV DNA load.

Effects of cytoplasmic transduction peptide-HBcAg18-27-Tapasin induced murine bone marrow derived dendritic cell maturation on T lymphocyte proliferation in vitro / 中华传染病杂志

Objective To observe the effects of cytoplasmic transduction peptide (CTP)-HBcAg18-27-Tapasin induced murine bone marrow-derived dendritic cell (DC) maturation on T lymphocyte proliferation in vitro,Methods Bone marrow derived DC isolated from BALB/c mice were cultured with recombinant granulocyte-macrophage colony-stimulating factor and recombinant interleutin (IL)-4 for 5 days followed by lipopolysaccharide added to induce DC maturation.10 μg/L CTP-HBcAg18-27-Tapasin,50 μg/L CTP-HBcAg18-27-Tapasin,10 μg/L CTP-HBcAg18-27 or RPMI-1640 were added into culture medium to induce DC maturation.DC phenotypes were analyzed by flow cytometry.The level of IL-12p70 in the supernatant was detected by enzyme linked immunosorbent assay.The proliferation of.T lymphocytes was performed by using cell counting kit-8 and intracellular cytokine of proliferative T cells were analyzed by flow cytometry.The means among groups were compared using one-way ANOVA and those between two groups were compared by least significant difference test.Results DC were cultured and induced successfully.The molecules on DC surface,such as CD80,CD86 and major histocompatibility antigen-Ⅰ were upregulated by CTP-HBcAg18-27-Tapasin.IL-12p70 level induced by 50 μg/L CTP-HBcAg18-27-Tapasin was (61.12±10.25) pg/mL,which was higher than those induced by 10 μg/L CTP-HBcAg18-27-Tapasin (50.43±10.42) pg/mL,10μg/L CTP-HBcAg18-27 (40.17±8.54) pg/mL and medium control (30.51±8.03) pg/mL (F=15.85,P=0.030 and 0.037).The proliferation of T lymphocytes induced by CTP- HBcAg18-27 -Tapasin was higher than control groups.The amounts of cytotoxic T lymphocyte (CTL) induced by 50 μg/L CTP-HBcAg18-27-Tapasin [(2.05±0.41) ％] and 10 μg/L CTP-HBcAg18-27-Tapasin [(1.06 ±0.10 )％] were both significantly higher than the 10 μg/L CTP-HBcAg18-27 group [(0.45±0.11)％] and medium group [(0.09±0.02)％,F=60.22,P=0.003].Conclusions CTP HBcAg18- 27 Tapasin could promote the differentiation and maturation of DC,and enhance the ability of DC stimulating T lymphocytes proliferation and increase CTL expression effectively.

Hepatitis B core antigen expression pattern predicts response to lamivudine therapy in patients with chronic hepatitis B / 대한간학회지

BACKGROUNDS/AIMS: Negative hepatitis B core antigen (HBcAg) staining in hepatocytes is indicative of viral replication by an active immune response. HBcAg is expressed mainly in the cytoplasm in patients with active hepatitis and hepatocyte regeneration, and mainly in the nuclei of hepatocytes in patients with minimal liver injury in the absence of hepatocyte regeneration. The aim of this study was to elucidate whether the existence and expression pattern of HBcAg predicts the response to antiviral treatment. METHODS: The study involved 58 patients with biopsy-proven chronic hepatitis B who were treated with lamivudine. Hepatitis B e antigen (HBeAg), antibody to HBeAg, hepatitis B virus DNA, and alanine aminotransferase in serum were recorded every 3 months. The inflammation grade and the fibrosis stage of chronic hepatitis were scored from 0 to 4 according to lobular inflammation, portal inflammation, periportal inflammation, and fibrosis. RESULTS: The 58 patients included 49(84%) HBcAg-positive patients, with HBcAg staining confined to the cytoplasm in 15(31%) and in both cytoplasm and nuclei in 34(69%). The grade of lobular inflammation and the total histology score were significantly higher in patients with cytoplasmic expression of HBcAg than in HBcAg-negative patients (lobular inflammation: 2.9 vs 2.1, P=0.02; total histology score: 12.2 vs 10.3, P=0.04). The virologic responses at 3, 6, 9, and 12 months differed significantly between the cytoplasmic and mixed expression groups (P<0.01). CONCLUSIONS: The expression pattern of HBcAg (including its possible absence) before initial therapy appears to predict the response to antiviral treatment.

Associations of Expressions of HBcAg and HBsAg with the Histologic Activity of Liver Disease and Viral Replication

BACKGROUND/AIMS: Subcellular localization of hepatitis B virus (HBV) core antigen (HBcAg) and HBV surface antigen (HBsAg) is known to be related to the activity of liver disease and the level of HBV replication. The aim of this study was to determine the correlation between histologic activity, viral replication, and the intracellular distributions of HBcAg and HBsAg. METHODS: We enrolled 670 patients with chronic hepatitis B who underwent liver biopsy at Bundang CHA hospital between 1997 to 2007. The data from medical records were reviewed retrospectively. RESULTS: The stage of fibrosis was higher (3.31+/-1.34 vs. 2.43+/-1.39, mean+/-SD, p0.05). CONCLUSIONS: These observations suggest that the histologic activity of hepatitis is higher and viral replication is lower in cHBcAg positive patients than in those with nHBcAg.

The Study of Expression HBcAg and HBsAg preS1 Epitope Peptide Fusion Protein / 中国生物工程杂志

To construct, express and purify fusion protein containing HBcAg and HBsAg preS1 epitope peptide for the purpose of investigating a novel HBV vaccine with both prophylactic and therapeutic functions. Using DNA recombinant technology, prokaryotic expression plasmid pBTcs1 expressing HBcAg and HBsAg pre-S1 epitope peptide fusion protein was constructed. After expressed in E.coli. HB101, the production BTcs1 was purified by sucrose density gradient ultracentrifugation and identified by SDS-PAGE, SEC, Western-blot and electron microscope. The results indicated that expression plasmid pBTcs1 was constructed successfully, and 20~25 mg purified BTcs1 fusion protein was obtained from 1L LB culture. Result of DOT-BLOT indicated that the distribution of BTcs1 was mainly in 30~50% sucrose, the purity of BTcs1 was greater than 95% by SDS-PAGE and SEC analysis. BTcs1 could probe with specific antibodies at 28 kDa by Western-blot, BTcs1 could also self assemble VLP by electron microscope analysis, its diameter was 30~34 nm approximately. The present study lay a foundation for further research functions and applications of BTcs1.

Relevancy between Liver Injury, Serum HBV-DNA, and Intrahepatic HBcAg in Young Male Chronic HBV Carriers / 대한소화기학회지

BACKGROUND/AIMS: Although the viral load is correlated with HBcAg, liver injury was not correlated to viral load in HBeAg positive patient. We aimed to study the inter-relationship of clinical parameters such as the level of HBV-DNA, the level of aminotransferase, intrahepatic expression of HBcAg and severity of histological liver damage in the young male chronic HBV carriers according to HBeAg status. METHODS: The study group included 85 young male patients (mean age: 19.8) with biopsy-proven chronic hepatitis B (HBeAg-positive group: n=60, HBeAg-negative group: n=25). RESUTLS: Serum levels of HBV-DNA and the expression of intrahepatic HBcAg in the HBeAg-positive group were significantly higher than in the HBeAg-negative (p<0.001), but fibrosis score was lower (p<0.01). Serum levels of HBV-DNA positively correlated with lobular activity, portal/periportal activity, biochemical activities in the HBeAg-negative group but negatively correlated in the HBeAg-positive group. There were no significant differences in histological activity according to the pattern of expression of intrahepatic HBcAg in both groups. The lobular activity correlated positively with biochemical activity in both groups, and portal/periportal activity correlated with biochemical activity only in the HBeAg-positive group. CONCLUSIONS: There are close correlations among liver injury, intrahepatic expression of HBcAg, and detectable HBV-DNA in the young male chronic HBV carriers with HBeAg-negativity, but in the HBeAg-positive group, the correlations are diversified.

A Study on Relationship between Serum HBV DNA and HBsAg , HBcAg Expression in the Hepatocytes in Chronic Hepatitis B Patients / 中国医师杂志

Objectives To study the relationship between serum HBV DNA and HBsAg, HBcAg expression in the hepatocytes in chronic hepatitis B(CHB) patients. Methods Quantitative polymerase chain reaction was used to assay the content of serum HBV DNA in 170 CHB patients. The expression of HBsAg and HBcAg in hepatocyctes was detected by immunohistochemical staining using liver biopsy. Results Although serum HBV DNA was negative, the expression of HBsAg in hepatocyctes was still observed in the CHB patients. In the CHB patients with lower level of serum HBV DNA, the positive rate of HBsAg expression in hepatocyctes was also lower (P

The Correlation Between The Histologic Activity and Fibrosis and The Distribution of Intrahepatic HBsAg and HBcAg in Patients With Chronic Hepatitis B / 대한간학회지

BACKGROUND/AIMS: The purpose of this study was to assess the correlation between histologic activity and fibrosis and the distribution of intrahepatic hepatitis B core antigen (HBcAg) and surface antigen (HBsAg) in patients with chronic hepatitis B. METHODS: 141 patients (M:F=141:27) with biopsy-proven chronic hepatitis B, abnormal liver function, and a positive HBV viral marker (serum HBeAg, serum HBV DNA) were enrolled. RESULTS: HBcAg was expressed in 96 of 141 patients (68.1%), nHBcAg in 23 (16.3%), cHBcAg in 58 (41.2%), and n-cHBcAg in 15 (10.6%). In the cases of HBsAg, 114 of 141 patients (80.9%) were expressed as cHBsAg, 2 (1.4%) as mHBsAg, and 16 (11.3%) as m-cHBsAg. The presence of intrahepatic HBcAg and HBsAg according to Gudat's classification was not correlated with activity and fibrosis. But the groups with nuclear expression of HBcAg revealed less inflammatory activity (grade, p=0.003), and less fibrotic stage (p = 0.002) than with cytoplasmic or no expression of HBcAg. HBsAg was not. CONCLUSIONS: These observations suggest that inflammatory activity and fibrosis of chronic hepatitis B are related to the presence of HBcAg in hepatocytes and the expression of HBcAg. This is a very important finding in hepatocytolysis.

An Immunohistochemical Study of the Expression of Hepatitis B Virus Antigens and p53 in Patients with Cutaneous Vasculitis Accompaied by Hepatitis B Surface Antigen-positive Hepatopathy / 대한피부과학회지

BACKGROUND: Cutaneous vasculitis associated with viral hepatitis seems to occur as a hypersensitivity reaction against the circulating viral antigens. Hepatitis B virus(HBV)-encoded X antigen(HBxAg) is known to participate in the carcinogenesis of hepatocellular carcinoma(HCC) by the inactivation of p53. However, HBxAg has been found in chronic infiammatory lesions without the overexpression of p53. Accordingly, not only EBsAg and HBcAg but also HBxAg may be involved in HCC-associated cutaneous vasculitis, regardless of the alteration of p53. OBJECTIVE: This study was conducted to investigate the expression of HBV-encoded antigens in cutaneous vasculitis accompanied by HBV hepatopathy. Additionally, we have compared the expression of 3 HBV antigens and p53 between vasculitic patients with HCC and in others showing HCC-non-associated vasculitis. METHODS: Immunohistochemically, we examined the expression of HBsAg, HBcAg, and HBxAg in the tissue specimens taken from the vasculitic lesions of the 33 HBsAg-positive enrolled patients with cutaneous vasculitis proven by skin biopsy. RESULTS: 1. The immunohistochemical positivity rate to HBsAg in vasculitic patients with HBV hepatopathy was 66.7% overall. It was 90% in HCC-associated vasculitic subjects and 56.5% in the vasculitic subjects without HCC, respectively. 2. We found the expression of HBxAg in 80% of the vasculitic subjects showing HCC. The vasculitic patients without HCC showed 17,3% of the positivity rate to HBxAg. 3. We could not find the overexpression of p53 in the vasculitic tissue specimens of the HCC patients without the cutaneous metastasis from primary HCC. CONCLUSION: HBsAg, HBcAg and HBxAg may participate in the pathogenesis of cutaneous vasculitis with HBV hepatopathy, regardless of tumorigenesis.

Persistent Negative Anti-Bc in Chronic Hepatitis B Virus Carrier: linical Review, HBcAg and Nucleotide Sequence of It's Epitope in HLA A2 Heterozygotes / 대한간학회지

BACKGROUND/AIMS: HBcAg is the most immunogenic HBV component and anti-Bc usually persists irrespective of ongoing liver disease or clearance of the virus in human. Therefore anti-Bc is considered as the most sensitive and occasionally the only marker of the HBV infection. Nevertheless, there are a few HBsAg carrier with persistent negative anti-Bc. The epitope which responds to HBcAg is recently defined in HLA A2 from acute viral hepatitis patient due to HBV. So we studied the clinical and laboratory features and nucleotide sequence of HBcAg corresponding to HLA A2 in the HBsAg carrier with persistent negative anti-Bc. METHODS: The subject of these study consists of eight HBsAg chronic carriers with persistent negative anti-Bc. We followed up the clinical features and serological markers of HBV infection and determined the amount of humoral immunoglobulin, HBV DNA and HBcAg when we performed the HLA class I typing and sequencing analysis of core of HBV. Control cases were selected from 3 HLA A2 heterozygote cases with chronic HBsAg carriers with anti-Bc. RESULTS: All subjects had the HBsAg persistently and good health conditions with normal ranges of aminotransferase and humoral immunoglobulin. One of them was converted to anti-Bc-ositive during follow-p period. The level of HBV DNA in serum was higher than 1.2 pg/mL in 7 of 8 chronic HBV carriers. There was a trend of differences between chronic anti-Bc negative carriers and converted one case to anti-Bc positive in the serum of HBcAg and HBV DNA(p=0.06). But strong positive correlation was observed between the amount of HBcAg and HBV DNA in sera. The core portion of HBV was amplified in 4 of 6 HLA A2 heterozygotes by single PCR. When sequenced the PCR products of the above 4 chronic anti-Bc negative HBV carriers and 3 control cases directly, there were no significant difference in the nucleotide and amino acid sequence at the HBcAg epitope which corresopond to class 1 HLA A2. CONCLUSIONS: Our results show that persistent anti-Bc negative chronic HBV carriers may be caused by large amounts of HBV DNA and HBcAg in their sera and not by variants of HBV. These suggested that active viral replication was going on, but are undetectable by the available commercial tests due to binding with excessive amount of HBcAg in the HBV carriers with persistent negative anti-Bc.

The Expression Rate and Pattern of HBcAg and HBsAg in the Hepatocytes According to the Histologic Activity of Cirrhosis

Since the discovery of hepatitis B virus as one of the causes of hepatitis, liver and hepatocellular carcinoma, many hepatitis B viral markers that appear in infected individuals have been discovered and many efforts to understand the relationship between the emergence of viral markers and the progression of hepatitis have been performed. Gudat (1975) compared the expression of HBcAg and HBsAg in various conditions and stages of hepatitis but the pattern of expression of viral markers and its significance have not been understood. Recently it was found by mierocytotoxicity assay that HBcAg might be the target of T lymphocytes. This study attempted to identify any correlation of the tissue expression rate and pattern of HBcAg and HBsAg with the histologic activity of 46 cases of liver cirrhosis using immunohistochemical staining. The expression rate and pattern of HBcAg and HBsAg in relation to the nodular size and positivity of serum HBeAg were also compared. The results were as follows; 1) The expression rate of HBcAg in the liver was 41.3% (19/46). and that of HBsAg was 67.4% (31/46). 2) The histologic activity of liver cirrhosis appeared to be correlated with the expression of HBcAg, especially cytoplasmic HBcAg. 3) The positivity of serum HBeAg was significantly higher in active liver cirrhosis. 4) There was no relationship between the tissue expression of HBsAg and the histologic activity of liver cirrhosis. relationship existed between the nodular size and expression rate and pattern of HBcAg and HBsAg. This study suggests that the tissue HBcAg, especially the cytoplasmic HBcAg is the most likely factor determining the histologic activity of liver cirrhosis, and that the cytoplasmic HBcAg may be the ultimate cause and target of most host immune response.

The Tissue Expression of HBsAg and HBcAg in Hepatocellular Carcinoma and Peritumoral Liver

To evaluate the tissue expression rate and pattenr of HBsAg and HBcAg in tumors and peritumoral livers, an immunohistochemical study was undertaken on 47 surgically resected hepatocellular carcinomas(HCCs). The results are as follows. 1. Patient's sera were positive for HBsAg in 40 cases(85.1%). In the remaining 7 cases, the tumor and peritumoral liver expressed neither HBcAg nor HbSaG, suggesting that they were caused by other etiologies than hepatitis B virus. 2. The peritumoral liver had HBsAg and HBcAg in 95.0% and 27.5% among the 40 cases, respectively. But the tumor expressed HBsAg in 50.0% and HBcAg in none. 3. The expression of HBsAg within the tumor and both HBsAg and HBcAg in the peritumoral liver tended to be more frequent in the pretreated cases before surgery. 4. Edmondson-Steiner grade IV tumors revealed a lower expression rate of HBsAg than the low grade tumors(p<0.05). Incases with cirrhosis at peritumoral tissues, HBcAg was less frequently found than in those without cirrhosis. The majority of tissue HBsAg and HBcAg was represented as groups of positive cells. These results suggest that, during the development and progression of HCCs, the HBcAg containing cells are repeatedly removed and the HBcAg negative cells are selected, because cellular expression of HBcAg is the target of host immune response.

Expression pattern of Hepatitis B Viral Core Antigen (HBcAg) and Surface Antigen (HBsAg) in Liver of the Inactive HBsAg Carriers

To understand better the complex natural course of HBV infection, the expression patterns of HBcAg and HBsAg in the liver of 51 inactive serum HBsAg carriers (24 CPH and 27 NPD) were studied by immunohistochemical methods. The inactive serum HBsAg carriers were devided into 3 groups by the following expression patterns of serum HBeAg/anti-HBe status and tissue HBcAg and HBsAg. Pattern A (18 cases) : HBeAg+, cHBcAg+ (94.4%), mHBsAg+ (61.1%), pATTERN B (14 cases) : anti-HBe+, nHBcAg+, cHBsAg+, Pattern C (19 cases) : anti-HBe+, HBcAg-, cHBsAg+ (89.5%). There were no significant differences between CPH and NPD, lthough the core free pattern was more common in the latter. The cHBcAg was expressed in 17 of 18 (94.4%) HBeAg seropositive cases but only one of 33 cases with serum anti-HBe, suggesting that the cHBcAg is intimately related to HBeAg. Since the inactive HBsAg carriers also expressed cHBcAg and/or mHBsAg, the necro-inflammatory activity of HBV infected liver is assumed to depend on the host immune response rather than their presence alone

Localization Forms and Clinical Implication of HBsAg、HBcAg in Liver Tissue with Chronic Hepatitis B / 昆明医科大学学报

The study reports on HBsAg and HBcAg in liver tissue samples of 36 patients with chronic hepatitis B were detected with ABC method. Among them 12 cases were HBsAg and/or HBcAg positive (32.4%). We found a high positive rate of replication signs of HBV with chronic active hepatitis indicating active replication of HBV. Also the extent of HBV replication was found parallel to that of active hepatopathy. Those with strong positive stain- ing always have middle or heavy chronic active hepatitis. In our studies active HBV replication and hepatopathy were observed. HBsAg in the liver mainly showed spread form of distribution and HBcAg mainly showed nuclear and cytoplasmic distribution, which was con- sistent with relevant publications. These results indicate that HBV replication is relevant to hepatopathy and its activity

Biologic Significance of Hepatocyte Hepatitis B Core Antigen Expression in Chronic Hepatitis B Virus Infection

To elucidate the biologic significause of hepatocyte B core antigen (HBcAg) expression and its relation to the natural course of hepatitis B virus (HBV) infection, we tried to correlate the patterns of HBcAg with the HBV replication state and with disease activity in 40 needle biopsies performed on hepatitis B surface antigen (HBsAg) carriers aged from 15 to 46 years. In 32 of 40 cases, HBcAg was present in the hepatocyte nucleus (nHBcAg), in the cytoplams (cHBcAg) or in both (mixed). Pure nHBcAg was seen only in minimal hepatitis, but a diffuse pattern of expression of cytoplasmic HBcAg and mixed cytoplasmic and nuclear expression of HBcAg were seen in active hepatitis. There was also a good correlation between liver HBcAg and serum HBeAg. Cases in which HBcAg expression were observed were positive for serum HBeAg (81%) and the cases negative for HBcAg were all positive for serum anti-HBe.

The humoral and cellular immune responses induced in H-2~b micee by nucleic acidvaccines encoding protein of hepatitis B virus / 中国免疫学杂志

Abstract Objective:To observe and confirm the specific humoral and cellurar immune responses induced by the nucleic acid vaccinesencoding HBV PreS2+S of subtypes adw and adr and encoding HBcAg in H-2b mice. Methods: Twenty-three C57BL/6 (H-2b) mice were ran-domly divided into four groups:group pJW4303(group P); group pJW4303/MHBs/adw(group W); group pJW4303/MHBs/adr(group R) andgroup pJW4303/MHBs/adr+pJW4303/HBc(group R+C) .The imunization method is that each mouse was injected(i.m) into the quadriceps femurs muscles with the corresponding 100?g(100?l) of plasmid DNA at 0,2,4 w.ELISA determined the level of anti-HBs and anti-HBc an-tibody in each mouse sera and the concentrations of IEN-? and IL-4 culture supernatant of spleen cells.Antigen specific proliferation of mousesplenocytes was measured by 3 H-TdR incorporating assay and the stimulation index(SI) was calculated.Results:After the last immunization,anti-HBs antibody had been detected in all mice of group W,group R and group R+C.The titers of anti-HBs were ranged from 22 329 to665.5 mU/ml.There are very significant differences between DNA vaccine groups (group W,group R,group R+C) and control group (groupP)(P0.05).Anti-HBs was first de-tected in group R+C.After the first immunization of nucleic acid vaccine of pJW4303/HBc,anti-HBc antibody appeared in all mice of groupR+C. Neither anti-HBs nor anti-HBc antibody appeared in all nice of group R+C.Neither anti-HBs nor anti-HBc was detected in group P.The stimulation index(SI) represent the capacity of antigen specific mouse proliferation of splenocytes. HBsAg-specific SIs of mouse splenocytesfrom group W,group R and group R+C is hipe than that in group P(P

DETECTION OF HBcAg WITH NITROCELLULOSE-ENZYME LINKED IMMUNO- SORBENT ASSAY (NC-ELISA) / 解放军医学杂志

A new ELISA method for the detection of HBcAg in serum has been established with nitrocellulose as the solid carrier.HBV-anti-HBs complex formed with patient's serum and horse anti-HBs was applied to the nitrocellulose membrane under vacuum, using a special apparatus, and then was fixed on the filter at room temperature over night. HBcAg was determined with HRP-anti-HBc after treatment with 3mol/L NaCNS. Comparison with the methods for serum HBc Ag previously described, this technique was simple in performance. Its sensitivity and specificity were satisfactory for clinical purpose. The results from the study of clinical specimens indicated that HBcAg was closely correlative to HBsAg, anti-HBc and HBV-PHSAR.