Mutation at codon 130 in hepatitis B virus (HBV) core region increases markedly during acute exacerbation of hepatitis in chronic HBV carriers

Mutations within T-cell or B-cell epitopes are suggested to have some influence on the clinical course of chronic hepatitis B virus (HBV) infection. To investigate the relationship between liver cell injury and heterogeneity of the HBV core gene, we focused on the sequence of codon 130, which is located on both T- and B-cell epitopes, and serially analyzed the proportion of mutant virus (core130Thr) to wild-type virus (core130Pro) during the exacerbation of chronic hepatitis B. Sera obtained serially from five HBV carriers who had exacerbation of hepatitis, and three asymptomatic HBV carriers (ASCs) with persistently normal serum aminotransferase (ALT) values were studied, using the restriction fragment length polymorphism (RFLP) method. Core130Pro predominated in the sera in the remission state, but core130Thr increased markedly in parallel with ALT elevation and decreased again after the ALT peak, followed by the predominance of core130Pro, in all the five patients. In one patient, the ratio of core130Thr/core130Pro (Thr/Pro) was more than 70% at the ALT peak. On the other hand, in sera from the three ASCs core130Pro always predominated, and no divergence was identified in the ratio of Thr/Pro. Our data suggest that codon 130 is one of the most important immunogenic regions in the HBV core gene and that elevation of Thr/Pro could be the result of immune selection. Received: January 21, 2000 / Accepted: October 6, 2000     

HBV携带者T细胞亚群状况与肝组织病理学对比

目的:研究HBV携带者的细胞免疫功能变化及其与血清HBVDNA,HBeAg的关系,并对T细胞亚群与肝组织病理学改变进行对比分析,探讨HBV携带及其所导致肝组织病理改变的机制.方法:应用流式细胞仪检测109例HBV携带者和40例健康对照外周血T细胞亚群百分率,并对其中28例行肝组织病理学检查,ELISA法检测HBV标志物,PCR法检测HBVDNA.结果:HBV携带者外周血CD3 、CD4 细胞百分率及CD4 /CD8 比值较正常对照组显著降低(67.2±9.0%vs71.0±3.5%,P<0.05;33.1%±6.6vs40.3±2.8%,P<0.001;1.10±0.36vs2.01±0.19,P<0.001),CD8 细胞百分率明显升高(33.8±8.4%vs20.2±1.9%,P<0.001).HBVDNA( )组及HBeAg( )组分别与HBVDNA(-)组及HBeAg(-)组比较,CD3 细胞无统计学差异,CD4 细胞显著降低(31.2±6.3%vs37.2±5.4%,P<0.001;31.0±6.0%vs35.8±6.5%,P<0.001),CD8 细胞明显升高(36.7±8.4%vs27.9±4.2%,P<0.001;37.3±8.4%vs29.5±6.0%,P<0.001),CD4 /CD8 比值明显降低(0.91±0.32vs1.35±0.26,P<0.001;0.89±0.30vs1.26±0.33,P<0.001).HBVDNA( )组肝病理组织学改变68.8%达到G1S1及以上程度,而HBVDNA(-)组仅为16.7%,二组间差异显著(c2=5.57,P<0.01).G1S1组CD3 细胞较G1S0组明显降低(F=2.919,P=0.047),CD4 细胞降低(P>0.05).G2S1组与G1S0组相比,CD3 细胞及CD4 细胞百分率有降低趋势,CD8 细胞百分率有升高趋势,CD4 /CD8 比值有明显降低趋势.结论:HBV感染可导致HBV携带者细胞免疫功能改变,HBVDNA复制增加可进一步加重HBV携带者的细胞免疫功能紊乱,并加重肝组织损害.     

Precore region mutation in hepatitis B virus genome in early stage of infection: A study in hepatitis B e antigen-positive young carriers

To determine when the precore mutation at the 83rd nucleotide occurs, leading to the formation of a stop codon in the hepatitis B virus genome in carriers, which would indicate the presence of antibody to hepatitis B e antigen (anti-HBe), we investigated this mutation by direct sequencing and subcloning in 22 young hepatitis B antigen (HBeAg) (+) carriers. These subjects were 7–17 years old and were found during a survey for hepatitis B surface antigen (HBsAg) in three elementary schools, a junior high, and a senior high school. None of these carriers had clinical manifestations, although one-third of them had elevated serum alanine aminotransferase levels. All were HBeAg-positive by radioimmunoassay (RIA), and 6 of them had preserved titers of anti-HBe at the same time. Precore mutations were found in 4 subjects (18.2%), with predominance of the wild type. Although 3 of these 4 had preserved titers of HBeAb, the other had no HBeAb titers. In an other 3 subjects with preserved titers of HBeAb, the precore mutation was not detected, even after the subcloning of viral DNA. The remaining 15 subjects with HBeAg showed no precore mutation. Subjects with ALT levels exceeding 100IU/l were all HBeAg-positive without the mutation. It was clear that the precore mutation itself occurred in the subjects at an early age during the course of infection. However, the chronological relationship between the emergence of the precore mutation and the onset of hepatitis requires further study.     

慢性HBV携带者肝组织Fas和Fas配体表达变化及其意义

目的：观察慢性HBV携带者（chronic asymptomatic HBV carrier,ASC）肝组织Fas和Fas配体（FasL）的表达和血清可溶性Fas（sFas）水平的变化。方法对120例ASC行肝穿刺,采用免疫组织化学法检测肝组织Fas和FasL表达,采用ELISA法检测血清sFas水平。结果120例ASC肝组织完全正常者仅占11%,轻度肝炎占59%,而中、重度肝炎占30%;表现正常的肝组织无Fas和FasL表达,轻度肝炎肝组织多无表达或仅限于界面炎症区的肝细胞及淋巴细胞阳性表达,而中、重度肝炎肝组织Fas和FasL多为阳性,Fas强阳性率分别达34.8%和69.2%,FasL强阳性率分别达30.4%和53.8%,均明显高于正常组和轻度炎症组（P〈0.05）,同时中、重度肝炎组间也存在显著性差异（P〈0.05）,其分布除界面炎症区外,肝小叶中也弥漫分布;肝组织表现正常的ASC血清sFas水平为（1.68±0.33） ng/L,而轻、中、重度肝炎患者则分别为（2.58±0.31）ng/L、（3.94±0.21）ng/L和（5.94±0.26）ng/L,均较正常组显著升高（F=218.01,P〈0.01）,各组间sFas水平也均具有统计学差异（P〈0.01）。结论 Fas和FasL介导的肝细胞凋亡在慢性HBV携带者肝组织病变中可能起到了重要作用。     

Variations of hepatitis B virus core gene sequence in Western patients with chronic hepatitis B virus infection

Heterogeneity of the hepatitis B virus (HBV) core gene has been reported to be associated with the presence of active liver disease in Japanese patients with chronic HBV infection. This study evaluated the significance of HBV core gene heterogeneity in Western patients with chronic HBV infection. The hepatitis B virus precore/core gene from 45 patients (inactive:active liver disease ratio 16:29) was amplified from serum by polymerase chain reaction (PCR). Gel electrophoresis was employed to detect large deletions. The PCR amplicons from 13 patients (all HBV serotype adw but with a different spectrum of liver disease) were cloned and sequenced. Hepatitis B surface antigen (HBsAg) serotypes were tested by enzyme immunoassay (EIA) and hepatic expression of HBV antigens was assessed by immunohistochemistry. The HBV core gene was amplified from the serum of all 45 patients. Three patients had mixed infection with both precore mutant and wild-type HBV and all three had active liver disease. No patient had a large deletion of the HBV core gene. Hepatitis B virus core gene sequence variations were more common in the midcore region and there was no difference in the number of silent and missense substitutions between those with inactive and active liver disease. There was no correlation between the nucleotide or encoded amino acid substitutions and the clinical and biochemical parameters, including the subsequent response to interferon-α therapy ( n =37) or hepatic HBV antigen expression. Variation of the HBV core gene was not found to be preferentially associated with active liver disease in Western patients with chronic HBV infection. The pattern of hepatitis B core gene variation is in accord with the genomic organization of HBV.     

经树突状细胞递呈的HBcAg特异性细胞毒T淋巴细胞的体外研究

目的　从人的外周血树突状细胞 (DC)的抗原递呈方面研究慢性乙型肝炎的发病机制 ,并诱导出针对HBcAg特异性的细胞毒T淋巴细胞 (CTL)。方法　取健康人DC和患者DC ,比较两组的抗原递呈功能是否存在差异。以HBcAg体外冲击致敏DC ,与自体T淋巴细胞共育 ,诱导出HBcAg特异性CTL ;以HepG2细胞为对照靶细胞 ,转染HBVDNA的HepG2 2 15细胞为靶细胞 ,分别测定CTL在效靶比为 2∶1、6∶1和 2 0∶1时对HepG2细胞的非特异性杀伤率及对HepG2 2 15细胞的特异性杀伤率 ,并比较患者组与正常组特异性杀伤率的差异。结果　患者DC的抗原递呈功能(10 99.2 6 7± 2 39.12 ,1374 .8± 36 4 .15 5 ,2 717.78± 15 89.72 )较健康人 (314 7.933± 72 6 .5 7,384 3.0 0 0±10 6 0 .85 ,5 4 86 .86 7± 1790 .6 4 )弱 ;健康组与患者组CTL对HepG2各效靶比的非特异性杀伤作用差异无显著性。健康组与患者组CTL对HepG2 2 15的特异性杀伤作用差异有显著性 ;患者组CTL的活性 (7.1± 4 .33,15 .6 8± 3.3,2 7.6 6± 4 .5 9)较健康组 (2 0 .76± 6 .0 8,33.97± 8.0 0 ,4 9.6 3± 9.4 8)弱。结论　用HBcAg体外负载患者DC ,能诱导出抗原特异性的CTL ,这些CTL能特异性地杀伤相应靶细胞。     

High risk of hepatitis B‐virus reactivation after hematopoietic cell transplantation in hepatitis B core antibody‐positive patients

We investigated the serological changes in hepatitis B virus (HBV)‐related markers in 55 and 26 hepatitis B surface antigen (HBsAg)‐negative patients undergoing allogeneic and autologous stem cell transplantation, respectively, over the past 4 yr. Five of the 17 allogeneic and one of the five autologous patients with pretransplant anti‐hepatitis B core antigen antibodies (anti‐HBc) were HBsAg‐positive after transplantation, whereas none of the patients negative for anti‐HBc were HBsAg‐positive in both groups. All patients who became HBsAg‐positive received steroid‐containing immunosuppressive therapy for chronic graft versus host disease (GVHD) or myeloma. Four of the six patients developed flare of HBV hepatitis, and two patients did not. One patient developed fulminant hepatitis treated with lamivudine and plasma exchange. Other five patients received entecavir from the detection of HBsAg. Although HBV‐DNA levels became below the limit of detection in all patients, HBsAg positivity remained in three patients after 6 months of treatment. We concluded that anti‐HBc positivity is a risk factor for reactivation of HBV after both autologous and allogeneic transplantation, and HBV‐related markers should be monitored regularly in these patients. We also stress the efficacy of pre‐emptive use of antiviral agents in controlling HBV replication and limiting hepatic injury due to reactivation of HBV in these patients.     

Immune responses in hepatitis C virus infection and mechanisms of hepatitis C virus persistence

Immune responses against hepatitis C virus (HCV) play a crucial role in the pathogenesis of chronic hepatitis C. HCV infection often persists and leads to chronic hepatitis and eventually cirrhosis. Accumulated data suggest that HCV proteins suppress host immune responses through the suppression of functions of immune cells, such as cytotoxic T lymphocytes, natural killer cells, and dendritic cells. They also suppress the type 1 interferon signaling system. The resulting insufficient immune responses against HCV lead to the sustained infection. The appropriate control of immune responses would contribute to the eradication of HCV and the improvement of hepatitis, but there are still many issues to be clarified. This review describes the scientific evidence to support these emerging concepts, and will touch on the implications for improving antiviral therapy.     

Comparison of pre-core/core hepatitis B virus region in liver tissue and serum from patients with chronic hepatitis B infection.

We examined the relationship between HBV-DNA isolated from the liver and from the serum in patients with various serological characteristics of chronic hepatitis B infection. Amplification and direct sequencing of the HBV pre-core/core region was carried out in 9 patients who were seropositive for HBsAg and HBV-DNA--4 HBeAg positive and 5 anti-HBe positive. Complete sequence identity was observed between HBV-DNA isolated from the serum and the liver in individual patients. In addition, shortened forms of the HBV core ORF were detected in patients with chronic active hepatitis, but not in patients with chronic persistent hepatitis.     

Amino acid substitutions in the S region of hepatitis B virus in sera from patients with acute hepatitis

Background: An increase in the number of acute hepatitis patients with hepatitis B virus (HBV) of non-indigenous genotypes may reduce the efficacy of vaccination against HBV. Methods: We have determined the amino acid (aa) sequences in the major hydrophilic region (MHR) in the S region of HBV in patients with acute hepatitis B and compared those with the ones from HBV strains used for the production of HBV vaccines commercially available in Japan. Results: Of 48 patients studied, 11 were infected with genotype A, 11 with genotype B and 26 with genotype C HBV. The aa sequences of the nine genotype A isolates were the same as the aa sequence of J02205 which is used for the production of one of the commercially available recombinant vaccines. The aa sequences of the 11 genotype B isolates differed from the aa sequence of J02205 in two or three amino acids. Of the26 genotype C isolates, 22 had the same aa sequence as X01587 which is used for the production of another recombinant vaccine. The remaining genotype C isolates had aa substitutions at aa131, which have a potential to alter the hydropathy and the three-dimensional structure of the MHR. The differences among the three current HBV vaccines in aa sequences in the MHR theoretically alter the hydropathy and three-dimensional structure. Conclusion: Our results suggest that the transmission of HBV isolates with different genotypes or with aa substitutions in the MHR might reduce the efficacy of currently available HBV vaccines in the protection of HBV infections.     

Hepatitis B e antigen negative chronic active hepatitis: hepatitis B virus core mutations occur predominantly in known antigenic determinants

Summary. In chronic hepatitis B virus (HBV) infection seroconversion from hepatitis B e antigen (HBeAg) to hepatitis B e antibody (HBeAb) may be followed either by remission of the disease with low-level viraemia, or by continuing inflammation with high-level viraemia. In both situations the virus may acquire a mutation in the precore sequence which prevents it from encoding HBeAg. We now show that the number of amino acid substitutions in the HBV core is low in viral sequences from patients with HBeAg positive chronic liver disease and HBeAg negative HBeAb positive patients in remission, but the frequency of substitutions is high in HBeAg, negative HBeAb positive patients with active liver disease. Furthermore we show that these substitutions cluster in the promiscuous CD4⁺ T-helper-cell epitope and in HBV core/e antibody binding determinants, but are not found in regions recognized by major histocompatability complex (MHC) restricted cytotoxic T lymphocytes. Sequential viral sequences from patients before and after HBeAg/HbeAb seroconversion shows that core mutations arise either at the same time or after the precore stop mutation which prevents the virus from encoding HBeAg. These results are consistent with the hypothesis that after clearance of HBeAg, mutations in regions of the virus recognized by CD4⁺ helper T cells and B cells allow persistence of the HBe negative virus in HBeAb positive patients with viraemia and active hepatitis.     

B cell gene signature with massive intrahepatic production of antibodies to hepatitis B core antigen in hepatitis B virus–associated acute liver failure

Hepatitis B virus (HBV)-associated acute liver failure (ALF) is a dramatic clinical syndrome due to a sudden loss of hepatic cells leading to multiorgan failure. The mechanisms whereby HBV induces ALF are unknown. Here, we show that liver tissue collected at the time of liver transplantation in two patients with HBV-associated ALF is characterized by an overwhelming B cell response apparently centered in the liver with massive accumulation of plasma cells secreting IgG and IgM, accompanied by complement deposition. We demonstrate that the molecular target of these antibodies is the hepatitis B core antigen (HBcAg); that these anti-bodies display a restricted variable heavy chain (V_H) repertoire and lack somatic mutations; and that these two unrelated individuals with ALF use an identical predominant V_H gene with unmutated variable domain (IGHV1-3) for both IgG and IgM anti-HBc antibodies, indicating that HBcAg is the target of a germline human V_H gene. These data suggest that humoral immunity may exert a primary role in the pathogenesis of HBV-associated ALF.     

Risk of alanine aminotransferase flare-up among asymptomatic hepatitis C virus RNA carriers: a 10-year follow-up study

AIMS: The aim of this study was to determine the cumulative rate of flare-up of serum alanine aminotransferase (ALT) level during a 10-year follow-up period, and characterize the clinical, virologic features in 120 hepatitis C virus (HCV) RNA-positive asymptomatic carriers with persistently normal ALT levels for 6 months. RESULTS: All flare-up cases occurred during the first 5 years of the present study, 27.4% of carriers showed ALT flare-up during this period, but none in the second half of the study. Multivariate analysis showed that C100-3 antibody (Ab) and anti-human T cell leukemia virus (HTLV)-I Ab were two independent and significant predictors of ALT flare-up in hepatitis C virus (HCV) RNA asymptomatic carriers (P = 0.04, P = 0.03, respectively). Liver biopsy was performed in 44 patients (11 with flare-up of ALT level, whereas 33 had normal ALT levels). Histological features of chronic hepatitis with lymphoid infiltration in the portal tracts were commonly observed in all specimens, and no differences were noted between the flare-up ALT group and the persistently normal ALT group. CONCLUSIONS: Our results indicated that flare-up of ALT levels in asymptomatic HCV-RNA carriers with normal ALT levels occurs during the first 5 years of diagnosis, and that the presence of C100-3 and anti-HTLV-I antibodies are good predictors of a transient rise in ALT.     

High levels of serum hepatitis B virus DNA in patients with 'anti-HBc alone': role of HBsAg mutants

It remains unclear how the detection of hepatitis B core antibody (anti-HBc) in the absence of hepatitis B surface antigen (HBsAg) and antibody (anti-HBs) should be interpreted and whether all patients with this pattern need to be tested for hepatitis B virus (HBV)-DNA. This study aimed at reassessing the significance of 'anti-HBc alone' in unselected sera referred to the clinical laboratory and determining whether significant HBV viraemia can be found in this setting. Of the 6431 patients tested for HBsAg, total anti-HBc and anti-HBs in a Paris hospital over a 1-year period, 362 (5.6%) had 'anti-HBc alone' (24.8% of anti-HBc-positive patients). Only 11 of the 362 sera (3.0%) were found to be false positive. One patient was in the resolving phase of acute hepatitis B. HBV-DNA was detected in 10 of 362 (2.8%) patients, using a commercial standardized assay (threshold: 350 IU/mL). Viral loads exceeded 10(4) copies/mL in 6 of 10 patients. Mutations in the HBsAg immunodominant region were identified in seven of the viraemic patients. HBsAg was detected in only two cases when retested by one of the latest, multivalent assays. Neither human immunodeficiency virus nor hepatitis C virus serostatus distinguished between patients with and without HBV-DNA. In conclusion, 'anti-HBc alone' should be considered a risk marker for a so-called 'false occult' HBV infection with significant viraemia. Indeed, results in this hospital population indicate that a small proportion of patients with 'anti-HBc alone' have high viral loads, revealing the occurrence of infection with HBV mutants that escape detection even by multivalent HBsAg assays.     

Concentrating missense mutations in core gene of hepatitis B virus

To elucidate the temporal relationship between liver damage and mutation(s) in hepatitis B virus core gene, serial sera from a progressive liver disease patient and an asymptomatic carrier were studied. By direct sequencing, missense mutations in the core gene were only found in serum from the progressive liver disease patient during the period with frequent exacerbation. Using methods of cloning and sequencing, missense mutations were also found in clones derived from the progressive liver disease patient at a relatively early phase, but strains with a missense mutation from earlier sera did not exist in sera of a later period. Furthermore, there was a tendency of concentrating missense mutations in clones derived from the progressive liver disease patient. These data suggested that missense mutations in the core gene that occurred at an earlier phase might evoke an immune response to eliminate mutated virus and that concentrating missense mutations during a phase of exacerbation might be a result of adaptive mutation.     

恩替卡韦治疗慢性乙型肝炎病毒携带者近期疗效观察

目的 观察恩替卡韦治疗慢性HBV携带者的近期疗效与安全陛,探讨慢性HBV携带者抗病毒治疗的临床意义. 方法 慢性HBV携带者47例,慢性乙型肝炎患者46例,诊断均符合“慢性乙型肝炎防治指南(2010年版)”.两组患者均给予恩替卡韦分散片0.5 mg/d口服治疗,观察两组患者治疗第4、12、24、48周血清学应答率、生物化学应答率与突破率的差异,并观察药物相关不良事件发生率.数据均使用SPSS17.0统计分析软件进行,分别采用t检验和x2检验.结果 第4、12、24、48周完全病毒学应答率:慢性HBV携带者组分别为14.9％、51.1％、76.6％和97.9％;慢性乙型肝炎患者组分别为,17.4％、63.0％、89.1％和100.0％,两组间各时间点比较,差异均无统计学意义.第4、12、24、48周部分病毒学应答率:慢性HBV携带者组分别为42.6％、57.4％、85.0％和100.0％;慢性乙型肝炎患者组分别为47.8％、65.2％、89.1％和100.0％,两组间各时间点比较,差异均无统计学意义.第4、12、24、48周HBeAg阴转率:慢性HBV携带者组分别为0、2.1％、4.3％和8.5％;慢性乙型肝炎患者组分别为4.4％、8.7％、13.0％和21.7％,两组间各时间点比较,差异均无统计学意义.第4、12、24、48周HBeAg血清学转换率:慢陛HBV携带者组分别为0、0、2.1％和6.4％;慢性乙型肝炎患者组分别为0、4.4％、10.9％和17.4％,两组间各时间点比较,差异均无统计学意义.HBsAg阴转率与血清学转换率,两组各观察时段均为0.慢性乙型肝炎患者组第4、12、24、48周ALT复常率分别为26.1％、65.2％、91.3％和97.8％.两组患者均无病毒学突破和生物化学突破病例.两组患者均未观察到肾毒性、骨髓抑制、横纹肌溶解或其他药物相关不良事件.结论 采用恩替卡韦分散片治疗慢性HBV携带者近期疗效好、安全.     

Clinical significance of antibody against hepatitis B virus core antigen in patients with hepatitis C virus‐related hepatocellular carcinoma

Purpose: We investigated the unsettled issue of whether seropositivity for antibody to hepatitis B core antigen (anti‐HBc) affects characteristics of hepatitis C virus (HCV)‐related hepatocellular carcinoma (HCC). Methods: Antibody status was determined by enzyme immunoassay in 243 patients with this cancer, and associations with clinicopathologic characteristics and outcome were analysed. Serum hepatitis B virus (HBV) DNA was determined by real‐time polymerase chain reaction. Results: Of 235 patients with unequivocal serologic status, 142 were seropositive and 93 were seronegative. Clinicopathologic characteristics and overall cumulative survival rates were comparable between the two groups. However, seropositivity tended to predict poor outcome for patients in Child class B or C (P=0.068), those in tumour‐nodes‐metastasis‐based stage 3 or 4 (P=0.081), those with tumours exceeding 25 mm (P=0.068), and those with a past history of clinical liver disease (P=0.088). Multivariate analysis identified serum albumin, portal vein tumour thrombosis, and tumour size as independent determinants of survival. Serum HBV DNA was below 1.7 log copies/ml in all 40 patients tested. Conclusions: Overall, the clinical features of HCV‐HCC were unaffected by seropositivity for anti‐HBc. Seropositivity tended to worsen prognosis for subgroup with poor hepatic reserve or advanced tumours.     

Comparison of complete sequences of hepatitis B virus genotype C between inactive carriers and hepatocellular carcinoma patients before and after seroconversion

Background Most patients who acquire chronic hepatitis B virus (HBV) infection by perinatal transmission become inactive carriers (IC) after hepatitis B e (HBe) antigen seroconversion, whereas some patients have persistent abnormal serum transaminase levels and develop hepatocellular carcinoma (HCC) in the anti-HBe-positive phase. The aim of this study was to investigate the HCC-related mutations of HBV. Methods Complete sequences of HBV were examined among eight IC and eight HCC patients infected with HBV genotype C before and after seroconversion. Results The frequency of the T1653 mutation tended to be higher among HCC patients after seroconversion (16.7% vs. 62.5%; P = 0.086). The prevalence of a basal core promoter double mutation (T1762/A1764) was high among both IC and HCC patients after seroconversion (83.3% vs. 87.5%; P = 0.825). Among the HCC patients, a pre-S deletion mutant was detected in 62.5% patients before seroconversion, and in 37.5% patients after seroconversion. The core deletion mutant was also detected in 50% of HCC patients only before seroconversion. Deletion mutants of the pre-S or core region before seroconversion were significantly associated with HCC patients (0% vs. 62.5%; P = 0.007, 0% vs. 50%; P = 0.021, respectively). Conclusions Our data showed a significant association of pre-S and core deletion mutants before seroconversion with HCC development. The T1653 mutation after seroconversion was frequently found in HCC patients infected with HBV genotype C. These results suggest that mutations may be predictive factor for development of HCC.