Core Promoter Mutations Research Articles

Association of hepatitis B virus mutations in basal core promoter and precore regions with severity of liver disease: an investigation of 793 Chinese patients with mild and severe chronic hepatitis B and acute-on-chronic liver failure

ObjectiveTo investigate the features of hepatitis B virus (HBV) basal core promoter/precore (BCP/PC) mutations and genotypes in a large number of mild/severe chronic hepatitis B (CHB-M/CHB-S), and acute-on-chronic liver failure (ACLF) patients and analyze the clinical implications of the virologic features.Patients and methodsSera of 793 (325 CHB-M, 170 CHB-S, and 298 ACLF) patients admitted to or who had visited Beijing 302 Hospital from January 2005 to December 2008 were collected and successfully amplified for the HBV BCP/PC and a 1225-bp-long S/Pol (nt 54–1278) gene regions. Biochemical and serological parameters and HBV DNA level were routinely performed. Viral DNA was extracted and subjected to a nested PCR. Genotypes/subgenotypes were determined based on complete genomic sequence or on analysis of the 1225-bp-long S/Pol-gene sequence. HBV genotyping was performed by direct PCR sequencing followed by molecular evolutionary analysis of the viral sequences. A P value of <0.05 (two-sided) was considered to be statistically significant.ConclusionsOur findings suggest that CHB patients infected with BCP/PC mutant viruses are more susceptible to severe hepatitis and ACLF than those with the BCP/PC wild-type virus and that ACLF patients with PC mutant viruses have an increased risk of death. As such, the HBV PC mutation is a potential predictive indicator of ACLF outcome.

Résultats de trois méthodes pour la détection de la mutation précore G1896A du virus de l’hépatite B chez les donneurs de sang français : PCR temps réel, séquençage et test Inno-LIPA

Aim To screen hepatitis B virus (HBV) genotypes and associated basal core promoter (BCP; T1762A/A1764) and precore (PC; A1896) mutations among the 100 HBV surface antigen (HBsAg) positive voluntary blood donors in France. Methods HBV genotypes were determined by using direct sequence analysis. Three methods were used to detect G1896A mutation: non-commercial real-time PCR (PCRTR°, line probe assay (InnoLiPA HBV PreCore, INNOGENETICS ®) and direct sequencing of precore gene. HBV viral load was quantified with two commercial real-time PCR (COBAS ® AmpliPrep/COBAS ® TaqMan ® HBV Test/Roche and Real Time HBV/M2000/Abbott). Results The mean age of donors was 30 (18–64). Patients were from Africa (42%), Europa (50%), and Asia (8%). HBV/D was the most predominant (37%) genotype followed by HBV/A (31%) and HBV/E (22%). PC and BCP mutants were found in 57% with Inno-LIPA HBV test and 59% with both PCRTR and sequencing methods. A significant difference in the viral load of blood donors with wild and PC mutants was observed with the Taqman Cobas real time PCR (3,19 Log 10 UI/ml versus 4,93 Log 10 UI/ml, p < 0.05). Precore phenotype determination was in agreement with the three PC mutation detection methods in 56% of cases. Conclusions Non-Caucasian genotype E was present in the French blood donors. PC mutation was more common than BCP mutations in this study. As HBV infected blood donors were more often asymptomatic carriers, we could speculate that the G1896A mutation may favour the asymptomatic state, supporting previous observations.

Microarray for Hepatitis B Virus Genotyping and Detection of 994 Mutations along the Genome

Genome analysis of hepatitis B virus (HBV) in patient sera is helpful for monitoring treatment. We developed an improved version of a DNA microarray to identify HBV genotypes and to detect mutations of interest in the S, Pol, Core, and X genes. It includes an automated software analysis of fluorescence values for simpler, more robust data interpretation. In this version, probes were added to identify genotype H, to analyze 155 additional positions, and to detect 561 additional polymorphisms. Sequences were added to the alignments to resolve hybridization problems due to natural polymorphisms in the vicinity of important codons. The duplex PCR protocol allowed whole-genome analysis in a single tube. An alternative nested-PCR protocol allowed genotyping and mutations in S and reverse transcriptase (rt) genes in patients with low viral loads, as demonstrated in patients with less than 400 HBV genome copies/ml. Reproducibility was high, with variation coefficients lower than 3%. Only 0.57% of 20,771 codons from 253 samples could not be identified. The concordance with Sanger sequencing for the identification of codons improved from 92.8% to 95.7% with the improved version. Concordance was higher than 91% for codons associated with resistance to lamivudine, emtricitabine, telbivudine, famciclovir, entecavir, and tenofovir with vaccine escape and for pre-Core mutants. Concordance was lower for adefovir resistance mutations (68.6%) and mutations in the basal core promoter (60.3%), probably because hybridization efficiency was affected by the low GC content of the probes. A concordance of 93.7% with sequencing for genotype identification was observed in 190 specimens, lower than that obtained with the first version, possibly due to mixed virus populations.

Hepatitis B virus-DNA level and basal core promoter A1762T/G1764A mutation in liver tissue independently predict postoperative survival in hepatocellular carcinoma

Hepatitis B virus (HBV) is a major etiological factor of hepatocellular carcinoma (HCC). However, the postoperative prognostic value of the virological factors assayed directly from liver tissue has never been investigated. To address this issue, 185 liver samples obtained from the noncancerous part of surgically removed HBV-associated HCC tissues were subjected to virological analysis. Assayed factors included the amount of HBV-DNA in the liver tissues; genotype; and the presence of the HBV precore stop codon G1896A mutation, basal core promoter A1762T/G1764A mutation, and pre-S deletions/stop codon mutation. All virological factors and clinicopathological factors were subjected to Cox proportional hazard model analysis to estimate postoperative survival. It was found that an HBV-DNA level >3.0 × 10(7) copies/g of liver tissue and the presence of the basal core promoter mutation independently predicted disease-free (adjusted hazard ratio 1.641 [95% confidence interval (CI) 1.010-2.667] and 2.075 [95% CI 1.203-3.579], respectively) and overall (adjusted hazard ratio 2.807 [95% CI 1.000-7.880] and 5.697 [95% CI 1.678-19.342], respectively) survival. Kaplan-Meier survival analysis indicated that in-frame, short stretch (<100 bp) pre-S deletions, but not large fragment (>100 bp) pre-S deletions, were significantly associated with poorer disease-free (P = 0.005) and overall (P = 0.020) survival. A hot deletion region located between codons 107 and 141 of the pre-S sequence was identified for the short stretch pre-S deletion mutants. The amount of HBV-DNA in liver tissue and the presence of the basal core promoter mutation were two independent predictors for postoperative survival in HCC. A short stretch pre-S deletion located between codons 107 and 141 was strongly associated with a poorer postoperative prognosis.

Hepatitis B genotype G and high frequency of lamivudine-resistance mutations among human immunodeficiencyvirus/hepatitis B virus co-infected patients in Brazil

In this study, we evaluated the hepatitis B virus (HBV) genotype distribution and HBV genomic mutations among a group of human immunodeficiency virus-HBV co-infected patients from an AIDS outpatient clinic in São Paulo. HBV serological markers were detected by commercially available enzyme immunoassay kits. HBV DNA was detected using in-house nested polymerase chain reaction and quantified by Cobas Amplicor. HBV genotypes and mutations in the basal core promoter (BCP)/pre-core/core regions and surface/polymerase genes were determined by sequencing. Among the 59 patients included in this study, 55 reported prior use of lamivudine (LAM) or tenofovir. HBV DNA was detected in 16/22 patients, with a genotype distribution of A (n = 12,75%), G (n = 2,13%), D (n = 1,6%) and F (n = 1,6%). The sequence data of the two patients infected with genotype G strongly suggested co-infection with genotype A. In 10 patients with viremia, LAM-resistance mutations in the polymerase gene (rtL180M + rtM204V and rtV173L + rtL180M + rtM204V) were found, accompanied by changes in the envelope gene (sI195M, sW196L and sI195M/sE164D). Mutations in the BCP and pre-core regions were identified in four patients. In conclusion, genotype G, which is rarely seen in Brazil, was observed in the group of patients included in our study. A high prevalence of mutations associated with LAM-resistance and mutations associated with anti-HBs resistance were also found among these patients.

Clinical and virological characteristics of chronic hepatitis B with concurrent hepatitis B E antigen and antibody detection

The concurrent detection of hepatitis B e antigen (HBeAg) and its corresponding antibody (anti-HBe) in patients with chronic hepatitis B virus (HBV) infection is well established but the clinical features remain poorly understood. Demographic information, clinical and laboratory data were collected from 1624 consecutive inpatient records of patients with chronic hepatitis B. Viral genotype, basic core promoter and precore mutations were determined by direct sequencing. In vitro HBeAg and anti-HBe binding experiments were conducted with three pairs of HBeAg-positive and anti-HBe-positive serum samples, which were mixed at variable ratios and incubated at 37°C for 3-24h. Of the 1624 chronic patients, 169 (10.4%) had concurrent HBeAg and anti-HBe positivity, and this was associated with intermediate age and HBV-DNA load, higher alanine aminotransferase level and more pronounced liver damage compared with HBeAg-positive or anti-HBe-positive patients alone. HBeAg and anti-HBe titres (median and interquartile range, S/CO) in the concurrent positive group were 4.2 (1.8-9.6) and 0.54 (0.27-0.72), which were closer to their respective cut-off values than those of HBeAg-positive or anti-HBe-positive groups alone. For the cases successfully sequenced, 110/134 (82.1%) harboured T1762/A1764 or/and A1896 mutants. The binding experiments showed that HBeAg and anti-HBe could be concurrently observed provided an optimal ratio (HBeAg to anti-HBe) was chosen. In antiviral treatment-naive patients, concurrence of HBeAg and anti-HBe was not uncommon, and such patients had profound liver disease. An optimal ratio between HBeAg and anti-HBe led to their concurrent detection when sera were tested by sensitive assays.

Hepatitis B virus genotypes and precore and core mutants in UAE patients

BackgroundKnowledge of the HBV genotype with which a patient is infected is crucial information for a physician to have when planning clinical treatment for that patient. Previous studies have suggested that there are possible differences in the pathogenicity and therapeutic response of different HBV genotypes. However, the prevalence of the various HBV genotypes and Precore and Core mutations is unknown in the UAE. Therefore, we sought to determine the prevalence of the different HBV genotypes in the UAE population.Methodology/Principal FindingsA total of 88 HBsAg-positive patients were included in the study.A method for genotyping and subtyping HBV by partial HBsAg gene sequencing using primers that are complementary to all known genotypes was used. Precore and core region of these viruses were also sequenced in 88 patients.HBV genotype D was the most prevalent (79.5%) genotype identified in our study population, followed by genotypes A (18.2%) and C (2.3%). The following subtypes were isolated: ayw2 (80.7%), adw2 (14.8%), and adw (2.3%). The HBV-DNA viral load was higher in HBeAg-positive patients than it was in patients who were HBeAg-negative. Precore mutants were found in 51 (58.0%) of 88 patients. Mutations in the basal core promotor were found in 22 (25.3%) of 88 patients.Conclusion/SignificanceHBV infection is a major health problem in the UAE, and while genotypes B and C are the most prevalent HBV genotypes in the Asian population, our study reveals that genotype D is the predominant genotype that is present in the UAE. More patients were HBeAg-negative than were HBeAg-positive in our study sample, which could be due to the duration of infection of the included patients. Additionally, the viral loads of the HBeAg-positive patients were higher those of the HBeAg-negative patients. Analysis of nucleotide 1858 showed presence of thymine in all patients with genotypes C, and D and in a few patients with genotypes A. This nucleotide was closely related to the presence of precore mutants. Mutations in the basal core promoter were found in 22 of 88 (25.3%) samples. These mutations were more frequent in patients infected with genotype A (37.5%) and not found in patients infected with genotype C.

The association of hepatitis B virus genotype and the basal core promoter mutation in Qidong, China

To investigate the prevalence of hepatitis B virus (HBV) genotypes and the association with hepatocellular carcinoma (HCC) or basal core promoter (BCP) mutation in Qidong, China. The whole genome of HBV or X gene sequences were obtained from serum samples of HBV infected patients by using PCR and direct sequencing methods. Phylogenetic tree was constructed to determine the genotypes or subgenotypes of HBV. According to the phylogenetic tree constructed from full-length sequence of HBV, genotype C2 was predominant in Qidong area. It was prevalent in 44 out of the 48 cases (91.7%), whereas genotype B2 only existed in 4 cases (8.3%). No other genotypes or recombinant types were found in Qidong patients. The result of genotyping based on X gene sequence confirmed the above observation. In a total of 182 samples, 169 (92.9%) showed genotype C2 and 10 (5.5%) showed genotype B2. There were 3 (1.6%) patients showed a coinfection with C2 and B2. The infection rate of genotype C in Qidong was significantly higher than that in neighboring city Shanghai (chi(2) = 12.252, P less than 0.01). There was no significant difference of genotype distribution between HCC and chronic hepatitis groups (P is more than 0.05). The frequency of T1762/A1764 double mutation in genotype C2 (70.3%) was significantly higher than that in genotype B2 (30.8%, P less than 0.05). The other two types of point mutation which also occurred in BCP, i.e. T1766 and A1768, were only seen in genotype C2. (1) Genotype C2 is the predominant genotype in Qidong, China. (2) There is no association between genotype C and HCC in Qidong. (3) Genotype C has a higher prevalence of BCP mutation than genotype B.

Genetic variations of hepatitis B virus and serum aflatoxin-lysine adduct on high risk of hepatocellular carcinoma in Southern Guangxi, China

Southern Guangxi area is one of the endemic areas for hepatocellular carcinoma (HCC) in China. This study evaluates the roles of genetic variations of hepatitis B virus (HBV) and aflatoxin B(1) (AFB(1)) exposure in the formation of HCC in this high-risk area. The study recruited 60 HCC patients and 120 age-, gender-, and residency-matched controls. HBV genotype and basic core promoter (BCP) mutations were determined by nested-PCR/direct sequencing. Serum AFB(1)-lysine adduct was measured by high performance liquid chromatography-fluorescence detection. HBV genotype C was predominant in 75.0% of cases and 84.2% of controls. The 1762(T)/1764(A) double mutations, 1753(V) mutations, and 1752(V) mutations were associated with HCC risk evidenced by the adjusted odds ratio (OR) [95% confidence interval (95% CI)] of 3.89 (1.40-10.77), 2.87 (1.49-5.49), and 5.96 (1.75-20.25), respectively. The adjusted OR (95% CI) was 6.94 (1.68-27.78) for subjects with 1762(T)/1764(A) double mutations and high AFB(1)-lysine adduct level; 2.01 (0.24-14.29), for those with only 1762(T)/1764(A) double mutations; and 4.26 (1.16-15.38) for those with only high AFB(1)-lysine adduct level, respectively. The adjusted OR was 5.13 (1.79-14.71) for subjects with 1753(V) mutations and high AFB(1)-lysine adduct level; 1.20 (0.47-3.08) for those with only 1753(V) mutations, and 2.28 (1.01-5.31) for those with high AFB(1)-lysine adduct level, respectively. These data confirmed the association of BCP mutations with HCC risk and the additive effects of 1762(T)/1764(A) double mutations and 1753(V) mutations with dietary AFB(1) exposure in this high-risk area for HCC.

Main mutations in the hepatitis B virus basic core promoter (A1762T/G1764A) before HBeAg loss are markers that identify patients who will require long‐term treatment

Some patients continue to have detectable HBV-DNA levels with liver disease progression after hepatitis B e antigen (HBeAg) loss. It is important to identify these patients, candidates for long-term treatment. To evaluate hepatitis B virus (HBV) genotype and the main mutations in the basic core promoter (BCP, A1762T/G1764A) and precore (G1896A) sequences as markers of persistent HBV-DNA after HBeAg loss. We analysed 60 serum samples from 20 Caucasian, HBeAg-positive, chronic hepatitis B patients, who lost HBeAg and were followed-up longitudinally. HBV genotype and precore and BCP mutations were determined before, at the time of, and after HBeAg loss. After HBeAg loss, eight (40%) patients continued to have undetectable HBV-DNA and 12 (60%) had persistent HBV-DNA (median level 4.7 log(10) copies/mL). The presence of BCP mutations prior to therapy was the only variable associated with persistently detectable viraemia (P = 0.017). Four patients with genotype A and no mutations in the BCP region experienced hepatitis B surface antigen (HBsAg) loss after a mean period of 35 months from baseline. Main BCP mutations in HBeAg-positive patients are useful markers to identify patients who will not have sustained virological suppression after HBeAg loss and therapy discontinuation and could benefit from long-term treatment.

T1653 mutation in the enhancer II region of the hepatitis B virus genome in southern African Blacks with hepatocellular carcinoma

An increased incidence of C-to-T1653 transversion (T1653) in the enhancer II region of the core promoter of hepatitis B virus has been reported in Japanese and Chinese patients with hepatocellular carcinoma infected with genotypes B or C of the virus, but little information is available in patients infected with other genotypes. To document the prevalence of T1653 in Black Africans with hepatocellular carcinoma, in whom genotype A is the dominant genotype and subgenotype A1 the dominant subgenotype, and to correlate its presence with other core promoter mutations previously described in association with T1653. The presence of the mutations was determined in 84 patients with hepatitis B virus-induced hepatocellular carcinoma and 50 matched asymptomatic carriers of the virus by extracting viral DNA from serum, amplification by polymerase chain reaction assay, and nucleotide sequencing. T1653 was not found significantly more often in the cancer patients with genotype A and subgenotype A1 than in the controls. An association was found not only between T1653 and T1762, A1764 and dual T1762/A1764 in the patients with hepatocellular carcinoma, but also in the asymptomatic carriers. T1653 mutation of hepatitis B virus does not occur more often in Black African patients with hepatocellular carcinoma with genotype A and subgenotype A1 than in asymptomatic carriers of the virus. No correlation specific to hepatocellular carcinoma was found between T1653 and other core promoter mutations in these patients. The presence of the T1653 mutation did not influence the e antigen status of the patients.

A comparison of hepatitis B viral markers of patients in different clinical stages of chronic infection

PurposeHepatitis B viral markers may be useful for predicting outcomes such as liver-related deaths or development of hepatocellular carcinoma. We determined the frequency of these markers in different clinical stages of chronic hepatitis B infection.Methods We compared baseline hepatitis B viral markers in 317 patients who were enrolled in a prospective study and identified the frequency of these tests in immune-tolerant (IT) patients, in inactive carriers, and in patients with either hepatitis B e antigen (HBeAg)-positive or HBeAg-negative chronic hepatitis or cirrhosis.Results IT patients were youngest (median age 27 years) and HBeAg-negative patients with cirrhosis were oldest (median age 58 years) (p = 0.03 to <0.0001). The male to female ratio was similar both in IT patients and in inactive carriers, but there was a male preponderance both in patients with chronic hepatitis and in patients with cirrhosis (p < 0.0001). The A1896 precore mutants were most prevalent in inactive carriers (36.4%) and HBeAg-negative patients with chronic hepatitis (38.8%; p < 0.0001), and the T1762/A1764 basal core promoter mutants were most often detected in HBeAg-negative patients with cirrhosis (65.1%; p = 0.02). Genotype A was detected only in 5.3% of IT patients, and genotype B was least often detected in both HBeAg-Positive patients with chronic hepatitis and cirrhosis (p = 0.03). The hepatitis B viral DNA levels were lowest in inactive carriers (2.69 log10 IU/mL) and highest in IT patients (6.80 log10 IU/mL; p = 0.02 to <0.0001). At follow-up, HBeAg-positive and HBeAg-negative patients with cirrhosis accounted for 57 of 64 (89.1%) liver-related deaths (p < 0.0001).ConclusionDifferences in baseline hepatitis B viral markers were detected in patients in various clinical stages of hepatitis B virus infection. HBeAg-positive and HBeAg-negative patients with cirrhosis accounted for the majority of the liver-related fatalities.

710 RISK OF NON-HODGKIN LYMPHOMA AND CHOLANGIOCARCINOMA IN TAIWANESE PAROUS WOMEN WITH CHRONIC HEPATITIS B VIRUS INFECTION: A NATIONWIDE COHORT STUDY

response and improved clinical outcome in HBeAg positive patients. In previous studies, age, gender, ALT, AFP, fibrosis, HBV-DNA, precore/core promoter mutation, HLA, and genotype are considered to be important factors for HBeAgSC. The aim of our study is to identify predicting factors for HBeAgSC within a year. Methods: From 1991 to 2005, liver biopsies were performed on 851 Hepatitis B s antigen positive patients in our hospital. From this group, 234 HBeAg positive patients were chosen to be subjects for this study. In 60 (26%) patients, HBeAgSC was observed within a year after liver biopsy, but in the remaining 174 (74%) patients, HBeAgSC was not observed. We compared the following factors between the two groups; age; gender; ALT; platelets; albumin; AFP; pathological features; HBeAg; HBV DNA; HBcrAg levels; mutations in the precore (G1896A) / core promoter (A1762T / G1764A) domain; and antiviral treatments. Results: Age (37 vs. 37: n.s.); gender (M/F) (43/17 vs. 118/56: n.s.); median ALT level (213 vs.125 IU/L: p = 0.004); median platelets level (171 vs. 186×103/ml: n.s.); median albumin level (4.1 vs. 4.1 g/dl: n.s.); median AFP level (12.0 vs. 6.0: p = 0.048); F0–1/2–4 (13/47 vs. 75/99: 0.043); cell infiltration in portal area 0–1/2–4 (12/48 vs. 59/115: 0.043); piecemeal necrosis 0–1/2–4 (20/40 vs. 91/83: 0.011); lobular inflammation 0–1/2–4 (15/45 vs. 80/94: p = 0.004); median HBeAg level (99 vs. 888 s/co: p< 0.000); median HBV DNA level (7.2 vs.8.0 log copies/ml: p< 0.000); median HBcrAg level (7.2 vs. 8.1 LogU/ml: p < 0.000), antiviral treatment (yes/no) (29/31 vs. 62/112: n.s.). Under multivariate analysis, HBeAg (<500 s/co), lobular inflammation (≥2), precore mutant, core promoter mutant were identified as independent factors for HBeAgSC. Especially, the patients with 2 favorable predictors (HBeAg and lobular inflammation) achieved HBeAgSC in 55% of cases. But, the patients without 2 favorable factors experienced HBeAgSC in only 6 % of cases. Conclusions: Low serum HBeAg level (<500) and severe lobular inflammation (≥2) are independent factors for HBeAgSC within a year. HBeAgSC could be predicted by evaluation for these two factors.

711 HEPATITIS B VIRUS (HBV) INFECTION IN BELGIUM: RESULTS OF THE BELGIAN ASSOCIATION FOR THE STUDY OF THE LIVER (BASL) REGISTRY OF 1421 HBSAG CHRONIC CARRIERS

response and improved clinical outcome in HBeAg positive patients. In previous studies, age, gender, ALT, AFP, fibrosis, HBV-DNA, precore/core promoter mutation, HLA, and genotype are considered to be important factors for HBeAgSC. The aim of our study is to identify predicting factors for HBeAgSC within a year. Methods: From 1991 to 2005, liver biopsies were performed on 851 Hepatitis B s antigen positive patients in our hospital. From this group, 234 HBeAg positive patients were chosen to be subjects for this study. In 60 (26%) patients, HBeAgSC was observed within a year after liver biopsy, but in the remaining 174 (74%) patients, HBeAgSC was not observed. We compared the following factors between the two groups; age; gender; ALT; platelets; albumin; AFP; pathological features; HBeAg; HBV DNA; HBcrAg levels; mutations in the precore (G1896A) / core promoter (A1762T / G1764A) domain; and antiviral treatments. Results: Age (37 vs. 37: n.s.); gender (M/F) (43/17 vs. 118/56: n.s.); median ALT level (213 vs.125 IU/L: p = 0.004); median platelets level (171 vs. 186×103/ml: n.s.); median albumin level (4.1 vs. 4.1 g/dl: n.s.); median AFP level (12.0 vs. 6.0: p = 0.048); F0–1/2–4 (13/47 vs. 75/99: 0.043); cell infiltration in portal area 0–1/2–4 (12/48 vs. 59/115: 0.043); piecemeal necrosis 0–1/2–4 (20/40 vs. 91/83: 0.011); lobular inflammation 0–1/2–4 (15/45 vs. 80/94: p = 0.004); median HBeAg level (99 vs. 888 s/co: p< 0.000); median HBV DNA level (7.2 vs.8.0 log copies/ml: p< 0.000); median HBcrAg level (7.2 vs. 8.1 LogU/ml: p < 0.000), antiviral treatment (yes/no) (29/31 vs. 62/112: n.s.). Under multivariate analysis, HBeAg (<500 s/co), lobular inflammation (≥2), precore mutant, core promoter mutant were identified as independent factors for HBeAgSC. Especially, the patients with 2 favorable predictors (HBeAg and lobular inflammation) achieved HBeAgSC in 55% of cases. But, the patients without 2 favorable factors experienced HBeAgSC in only 6 % of cases. Conclusions: Low serum HBeAg level (<500) and severe lobular inflammation (≥2) are independent factors for HBeAgSC within a year. HBeAgSC could be predicted by evaluation for these two factors.

Level of Hepatitis B Virus DNA in Inactive Carriers With Persistently Normal Levels of Alanine Aminotransferase

Little is known about the level of hepatitis B virus (HBV) DNA in individuals with chronic, inactive HBV infections. Patients who test positive for the antibody to hepatitis B e antigen (anti-HBe) and have normal levels of alanine aminotransferase for more than 10 years have a low risk of HBV reactivation and are considered to be inactive carriers. We investigated HBV DNA levels in inactive carriers and identified factors that correlated with this state among anti-HBe-positive carriers with HBV DNA levels of 10(4) copies/mL or greater (5.26 copies/mL = 1 IU/mL). HBV DNA levels were assayed in 250 inactive carriers with persistently normal alanine aminotransferase levels for more than 10 years. Clinical and virologic features were compared between inactive carriers (with HBV DNA levels > or =10(4) copies/mL) and age-matched patients with HBe antigen-negative chronic hepatitis (controls, n = 90). The median level of HBV DNA among inactive carriers was 3.70 log(10) copies/mL (range, undetectable to 5.98 log(10) copies/mL). Ninety (36%) had levels of 10(4) copies/mL or greater. Compared with control patients, significant differences of inactive carriers included sex (more female patients), lower HBV DNA levels, and lower prevalence of genotype C virus and the basal core promoter mutation T1762/A1764. The prevalence of the precore mutation A1896 was similar between groups. Multiple logistic regression analyses identified male sex, HBV DNA levels greater than 10(5) copies/mL, and the basal core promoter mutation as independent factors that correlated with active disease. Nearly 40% of inactive carriers had HBV DNA levels of 10(4) copies/mL or greater. Female sex, HBV DNA levels of 10(4) to 10(5) copies/mL, and wild-type basal core promoter correlated with inactive carrier state.

Novel mutations in the calreticulin gene core promoter and coding sequence in schizoaffective disorder

We have recently reported the first case of mutation in the core promoter sequence of the human calreticulin gene in a family case of schizoaffective disorder. Remarkably, this gene coincides with a region of suggested linkage at 19p13.2, identified in a whole genome scan [Hamshere et al. (2005); Arch Gen Psychiatry 62;1081-1088]. The identified mutation was located at the conserved position -48 from the transcription start site, and was shown to be of functional effect, resulting in the aberrant expression of the gene. Following screening of the gene in 60 independent cases of schizoaffective disorder, we report novel germ-line mutations at positions -205 C > T and the conserved exon 5 (c: 682 C > T, pro228ser) in two unrelated cases of schizoaffective disorder. These mutations were disease-specific, and as for the -48 G > C mutation, neither was detected in a control population of 370 individuals, indicating a contribution of 3.17% in this sample series. To our knowledge, this is the first instance of disease-specific mutations in schizoaffective disorder, which warrants systematic screening of the regulatory and coding regions of the calreticulin gene in this disorder.

Molecular evolutionary analysis and mutational pattern of full‐length genomes of hepatitis B virus isolated from Belgian patients with different clinical manifestations

Molecular evolutionary patterns of 62 HBV full-length genomes obtained from Belgian patients were characterized. Phylogenetic analysis revealed diverse HBV subgenotypes including A2 and A6 (46.8%), D1-D4 (38.8%), E (9.7%), C1 (1.6%), and B2 (1.6%). The study population consisted of patients with different ethnic origin (Caucasian, Turkish, Asian, Arab, and African). One HBV D/C recombinant isolate was identified, which encoded subtype adw2. An HBV subgenotype D4 with an aberrant subtype ayw4 was detected. Although none of the genotypes was associated with a specific disease outcome, several nucleotide substitutions, deletions and insertions were observed within the HBV preS1/S and X genes, particularly among patients with active chronic hepatitis B infection and patients with cirrhosis. Within the immunological domain of the HBsAg gene, the most frequent substitutions were sT125M and sT118A. High rates of precore and basal core promoter mutations were detected in patients infected with genotype D of HBV. Almost half of the patients who received lamivudine therapy for at least 1 year had HBV variants associated with lamivudine drug resistance. In conclusion, the most common HBV genotypes in West Europe (A and D) also prevail in Belgium. The highest degree of genetic diversity was detected in HBV genotype D. In addition, this study reveals the circulation of exotic HBV genotypes B, C, and E in Belgium. J. Med. Virol. 82:379-389, 2010. (c) 2010 Wiley-Liss, Inc.

Features and clinical implications of hepatitis B virus genotypes and mutations in basal core promoter/precore region in 507 Chinese patients with acute and chronic hepatitis B

Background The association of hepatitis B virus (HBV) genotypes and basal core promoter (BCP) and precore (PC) mutations with the clinical characteristics is increasingly recognized. Objective To investigate virologic features and clinical implications of HBV genotypes, BCP and PC mutations between large-size patients with acute hepatitis B (AHB) and chronic hepatitis B (CHB). Study design One hundred and eighty-two AHB patients and 325 CHB patients were investigated. HBV genotypes and BCP/PC mutations were determined by direct sequencing. Mutations at 10 interested sites of the BCP/PC region were compared between the two groups of patients. Results AHB patients had a significantly higher ratio of genotype B to C than CHB patients (37.4–62.6% vs. 16.6–83.4%, P < 0.001). The prevalence of BCP/PC wild-type virus was 60.4% in AHB patients in contrast to 28.9% in CHB patients. Significantly lower prevalence of A1762T, G1764A, G1896A, and G1899A but higher prevalence of T1758C was found in AHB patients. Interestingly, T1758C and A1762T/G1764A appeared mutual restraint. Genotype B virus had lower BCP mutation frequency and similar PC mutation frequency compared to genotype C virus. AHB patients with BCP/PC mutant virus had higher viral load, whereas CHB patients with BCP/PC mutant virus had lower viral load and elevated alanine aminotransferase, in comparison with those with the wild-type virus. Conclusion Patients with genotype B virus, BCP/PC wild-type virus or T1758C mutant virus were more susceptible to develop AHB, whereas high prevalence of the BCP/PC mutations was associated with CHB development.

Response to Adefovir Depends on Mutation Patterns in Precore Region, Basal Core Promoter and Reverse Transcriptase, and On-Treatment Responses in Lamivudine-Resistant Chronic Hepatitis B Patients

Objective: In vitro studies showed that mutations in the basal core promoter (BCP) or precore (PC) region restore the replication inefficiency of the lamivudine-resistant mutant. The aim of this study was to clarify the effect of molecular characteristics on the antiviral response to adefovir in patients with lamivudine-resistant chronic hepatitis B (CHB). Methods: Sixty-six lamivudine-resistant patients who were treated with adefovir monotherapy were studied. Sequences of BCP, PC region and reverse transcriptase were determined before adefovir therapy. In patients with virologic breakthrough, reverse transcriptase sequencing was performed. Results: The cumulative probabilities of virologic response were 23.3, 46, 52.7 and 59.5% at years 1, 2, 3 and 4, respectively. PC mutation, the absence of compensatory mutations (rtL80I/V or rtV173L), and a decrease in serum hepatitis B virus (HBV) DNA by 3 log or greater at 6 months were independent predictors of virologic response. The cumulative probabilities of virologic breakthrough were 0, 12.9, 30.7 and 44.5% at years 1, 2, 3 and 4, respectively. BCP mutation and a less than 3 log decrease in serum HBV DNA at 6 months were 2 independent risk factors for virologic breakthrough. Conclusion: Response to adefovir depends on mutation patterns in the BCP, PC region and reverse transcriptase, and on-treatment decreases in serum HBV DNA in lamivudine-resistant CHB patients.

Drug-resistant and immune-escape HBV mutants in HIV-infected hosts

HIV-HBV-coinfected patients require optimal control of viral replication in order to prevent the development of severe comorbidities, such as liver cirrhosis and hepatocellular carcinoma. The genetic diversity of HBV is a poorly investigated factor of such viral replication in HIV-infected hosts. HBV genome diversity can be differentiated into two major aspects: genotypic and phenotypic. Genotypic diversity is more related to the natural history of HBV infection and genotypes are mostly determined by geographical origin of the hosts. Phenotypic diversity arises from attempts to escape from host immune surveillance (that is, precore, core and basal core promoter mutants), selection resulting from the use of treatments with weak genetic barrier (that is, pol mutants), exposure to hepatitis B immunoglobulin (that is, 'immune-escape' S gene mutants) or treatment-induced mutations from overlapping genes (that is, pol mutants inducing 'vaccine-escape' S gene mutants). pol mutations typically lead to uncontrolled viral replication, whereas S gene mutations can significantly alter hepatitis B surface antigen synthesis and reduce binding to antibodies, which renders individuals who are vaccinated or cured of HBV infection susceptible to infection. For patients coinfected with HIV, hepatitis B treatment options that aim to reduce the risk of HBV mutations from emerging must be seriously considered, not only from clinical but also public health perspectives.