Hepatitis B Virus DNA Load Research Articles

Use of tumor necrosis factor α inhibitors in hepatitis B surface antigen-positive patients: a literature review and potential mechanisms of action

As a class, tumor necrosis factor (TNF)-alpha inhibitors have provided clinicians significant control over chronic inflammatory diseases. With their widespread use has come the emergence of new side effects such as the reactivation of latent infections. One such infection that may reactivate is the hepatitis B virus (HBV). It is currently unknown if HBV reactivation is a class effect or attributable to a particular TNF-alpha inhibitor. To answer this question, a comprehensive literature review to identify trends in related cases was performed. A systemic literature review was performed using the PubMed and Medline databases (1996 to January 2010) searching for the index term "Hepatitis B" combined with the terms "tumor necrosis factor," "TNF-alpha inhibitors," "etanercept," "adalimumab," "certolizumab," and "golimumab." All relevant articles in English were reviewed, and secondary references of interest were also retrieved. Thirty-five cases with hepatitis B surface antigen (HBsAg) positivity known prior to initiation of TNF-alpha inhibitors were identified. Infliximab was used in 17 cases, etanercept in 12 cases, and adalimumab in 6 cases. All six cases of clinically symptomatic hepatitis were associated with infliximab therapy. Infliximab was associated with the most cases of greater than 2-fold increase in alanine aminotransferase (six of nine cases) and greater than 1,000-fold increase in HBV DNA load (three of four). The two deaths reported occurred with infliximab therapy. Potential mechanisms of action for the reported observations include differences in molecular design, route of administration, and potency in clearing TNF-alpha. In patients with a positive HBsAg prior to starting a TNF-alpha inhibitor, infliximab has the most reported cases associated with HBV reactivation. While such reactivation may be due to a variety of reasons, clinicians prescribing TNF-alpha inhibitors to HBsAg-positive patients should consider prophylactic antiviral therapy and close monitoring for any clinical or serological evidence of hepatitis.

Core Antigen Expression Is Associated with Hepatic Necroinflammation in e Antigen-Negative Chronic Hepatitis B Patients with Low DNA Loads

Intrahepatic hepatitis B virus (HBV) core antigen (HBcAg) is a hallmark of viral replication in hepatitis B virus e antigen (HBeAg)-positive chronic hepatitis B (CHB). The aim of this study was to evaluate the role of HBcAg in HBeAg-negative CHB. One hundred six HBeAg-negative CHB patients who underwent ultrasonographically guided liver biopsy were reviewed for their HBV DNA load and clinical and histological data. Factors associated with the expression of intrahepatic HBcAg were analyzed. Among the patients, 35 (33%) were positive for HBcAg by immunohistostaining. In patients whose HBV DNA loads were higher than 10(7) copies (cp)/ml, nearly one-half (52%) had detectable HBcAg. Compared with HBcAg-negative patients, HBcAg-positive patients had higher serum alanine transaminase (ALT) and HBV DNA levels and more-severe hepatic necroinflammation. High serum ALT level (>160 U/liter) and HBV viral load were the determinants of HBcAg expression in multivariate analysis. Large amounts of HBcAg expression were frequently detected in patients with high DNA loads, and the patterns of HBcAg distribution were not related to histological activity or HBV DNA levels. In patients with lower HBV DNA loads, the expression of HBcAg was the key factor associated with active hepatic necroinflammation (hazard ratio = 11.25; 95% confidence interval [CI], 1.42 to 89.26; P = 0.022). In conclusion, the expression of HBcAg is not frequent in HBeAg-negative CHB. The expression of intrahepatic HBcAg indicates active hepatic necroinflammation, even in patients with low HBV DNA load. Both HBV viral load and HBcAg expression have implications in the pathogenesis of HBeAg-negative CHB.

Polymerase region mutations and hepatitis B virus genotypes in chronic hepatitis B patients with poor response to lamivudine

To investigate the mutations in Polymerase region and hepatitis B virus (HBV) genotypes in chronic hepatitis B patients with poor response to Lamivudine treatment. 631 chronic hepatitis B patients with poor response to Lamivudine were recruited in this study. Real-time PCR and DNA sequencing were used to determine HBV genotypes; direct sequencing was performed to detect mutations, and real-time PCR was used to quantify HBV DNA load. Mutations in polymerase region were investigated in different HBV genotypes. 272 patients were infected with HBV of genotype B, and 359 patients were infected with HBV of genotype C. The mean age of patients infected with HBV of genotype C (39.1+/-11.4 years old) were significant higher than that of patients infected with HBV of genotype B (33.7+/-9.7 years old) (t = -6.55, P less than 0.01). The patients infected with HBV of genotype C had relatively higher HBV DNA load [(5.96+/-1.22) log10 copies/ml] than the patients infected with HBV of genotype B [(5.58+/-1.21) log10 copies/ml] (t = -2.01, P less than 0.05). The overall incidence rate of A181V/T mutation in genotype C (5.3%) was significantly higher than that in genotype B (0.4%) (x2=12.23, P less than 0.01), but the incidence rate of M204I/V, L180M, T184A/G/I/S, S202G/I and V173L mutations was not significantly different between genotype B and C (each P more than 0.05). M204I mutation in genotype B (20.6%) was more frequent than that in genotype C (13.9%) (x2=4.91, P less than 0.05). The Lamivudine resistance mutations were not significantly different between genotype B and genotype C (x2 = 0.00, P more than 0.05). The incidence rate of lamivudine resistance mutation is not significantly different between genotype B and genotype C, but patients infected with HBV of genotype C have higher HBV DNA load than patients infected with HBV of genotype B.

Influential factors of prognosis in lamivudine treatment for patients with acute‐on‐chronic hepatitis B liver failure

Chronic hepatitis B virus (HBV) infection is a major global health issue, and the prognosis of patients with HBV-associated acute-on-chronic hepatic failure (ACLF) is extremely poor. In this study, the efficacy of lamivudine was investigated in patients with ACLF. The effects of HBV DNA load and its related factors on the prognosis were also further explored. A matched retrospective cohort study using data on ACLF patients derived from our hospital database was conducted. One hundred and thirty patients receiving lamivudine were selected into the lamivudine treatment group with another 130 without lamivudine treatment studied as control. They were matched for sex, age and imaging finding with the lamivudine treatment group. All the patients were followed up for 3 months and the survival rates were compared. The influential factors on the mortality were studied by the Cox proportional hazards model. The cumulative survival rates of patients in the lamivudine group were higher than those of the control group (chi(2) = 9.50, P = 0.0021). The mortality of patients in the high virus load group (71/95, 74.7%) was higher than that of those in the low virus load group (15/29, 51.7%) (chi(2) = 5.536, P = 0.019). For patients with a Model for End-Stage Liver Disease (MELD) score of 20-30 by week 4, the mortality of those with HBV DNA that was undetectable or declined for more than 2 log(10) (2/12, 16.7%; 18/40, 45.0%) was lower than that of those with a less than 2 log(10) decline (18/23, 78.3%) (chi(2) = 10.106, P = 0.001). In the Cox proportional hazards model, for patients with a MELD score of 20-30, treatment method (P = 0.002), pretreatment HBV DNA load (P = 0.007) and decline of HBV DNA load during therapy (P = 0.003) were independent predictors; for those with a MELD score of above 30, MELD score (P = 0.008) was the only independent predictor. Lamivudine can significantly decrease the 3-month mortality of patients with a MELD score of 20-30, and a low pretreatment viral load and rapid decline of HBV DNA load are good predictors for the outcome of the treatment.

Serum Hepatitis B Surface Antigen and Hepatitis B E Antigen Titers: Disease Phase Influences Correlation With Viral Load and Intrahepatic Hepatitis B Virus Markers

Although threshold levels for hepatitis B surface antigen (HBsAg) and hepatitis B e antigen (HBeAg) titers have recently been proposed to guide therapy for chronic hepatitis B (CHB), their relationship to circulating hepatitis B virus (HBV) DNA and intrahepatic HBV replicative intermediates, and the significance of emerging viral variants, remains unclear. We therefore tested the hypothesis that HBsAg and HBeAg titers may vary independently of viral replication in vivo. In all, 149 treatment-naïve CHB patients were recruited (HBeAg-positive, n = 71; HBeAg-negative, n = 78). Quantification of HBeAg and HBsAg was performed by enzyme immunoassay. Virological characterization included serum HBV DNA load, HBV genotype, basal core promoter (BCP)/precore (PC) sequence, and, in a subset (n = 44), measurement of intrahepatic covalently closed circular DNA (cccDNA) and total HBV DNA, as well as quantitative immunohistochemical (IHC) staining for HBsAg. In HBeAg-positive CHB, HBsAg was positively correlated with serum HBV DNA and intrahepatic cccDNA and total HBV DNA (r = 0.69, 0.71, 0.76, P < 0.01). HBeAg correlated with serum HBV DNA (r = 0.60, P < 0.0001), although emerging BCP/PC variants reduced HBeAg titer independent of viral replication. In HBeAg-negative CHB, HBsAg correlated poorly with serum HBV DNA (r = 0.28, P = 0.01) and did not correlate with intrahepatic cccDNA nor total HBV DNA. Quantitative IHC for hepatocyte HBsAg confirmed a relationship with viral replication only in HBeAg-positive patients. The correlation between quantitative HBsAg titer and serum and intrahepatic markers of HBV replication differs between patients with HBeAg-positive and HBeAg-negative CHB. HBeAg titers may fall independent of viral replication as HBeAg-defective variants emerge prior to HBeAg seroconversion. These findings provide new insights into viral pathogenesis and have practical implications for the use of quantitative serology as a clinical biomarker.

Th1 and Th2 Immune Response in Chronic Hepatitis B Patients during a Long-Term Treatment with Adefovir Dipivoxil

Adefovir dipivoxil treatment has significantly improved the outcome of chronic hepatitis B virus (HBV) infection. However, it remains largely unknown how immune system responds to the treatment. Chronic HBV patients were treated with adefovir dipivoxil and examined for serum HBV DNA loads, cytokines, and T helper (Th1) and 2 (Th2) cytokine producing T cells during 104 weeks of the treatment. Th1/Th2 cytokines producing T cells were significantly lower in chronic HBV patients as compared to normal individuals. Adefovir dipivoxil treatment led to the increase of Th1/Th2 cytokines producing T cells and serum cytokine levels in association with the decline of HVB DNA load. In contrast, Th1/Th2 cytokines producing T cells remained lower in one patient detected with adefovir dipivoxil resistant HBV A181T/V mutation. This study has established inverse correlation of the increase of Th1/Th2 immunity and the decline of HBV DNA load in chronic HBV patients during adefovir dipivoxil treatment.

Risk of Hepatitis B Virus (HBV) Reactivation and the Role of Anti-Viral Prophylaxis in Multiple Myeloma Patients with HBV Infection in the Era of Novel Therapies.

Abstract 3882Poster Board III-818 IntroductionPatients with hepatitis B virus (HBV) infection, defined by the presence of HBV surface antigen (HBsAg), have an increased risk of HBV reactivation when they are on immunosuppressive treatment for multiple myeloma (MM). Although there is no guideline for MM patients with HBV infection, current lymphoma guidelines do recommend that these patients should receive antiviral prophylaxis during and after chemotherapy. Of late, the advent of bortezomib in the management of MM has resulted in a high reported incidence of variecella-zoster reactivation. The risk of HBV reactivation in MM patients with HBV infection undergoing treatment has not been previously studied. As HBV infection is endemic in Asia, we sought to evaluate the prevalence of HBV infection in our patients, the incidence of its reactivation especially in patients receiving bortezomib and the role of anti-viral prophylaxis. MethodsPreviously untreated MM patients diagnosed from 2000-2008 who were tested for HBsAg in our institution were included. Hepatitis attributable to HBV reactivation was defined as an increase in HBV DNA levels of tenfold, or an absolute increase greater than105 copies/ml in the HBV DNA level. HBV infected patients were prospectively followed. 33% of all patients have been exposed to bortezomib, while 26% received high dose therapy with autologous stem cell transplantation (HDT/ASCT). Results243 untreated MM patients were identified. The prevalence of HBV infection is 5.8% (14/243). 6 (43%) HBV infected patients had detectable HBV DNA viral load (>3 log) at baseline. All 6 patients had normal baseline liver function tests and received lamivudine prophylaxis. All 14 HBV infected patients went on to receive systemic therapy for MM, with continual monitoring of HBV DNA viral load and liver enzymes for viral reactivation. 4 patients with undetectable HBV DNA load did not receive anti-viral prophylaxis. Of these 14 patients, 3 (21%) who had been on lamivudine prophylaxis had reactivation of the virus, with 1 dying from it, and 1 having emergence of a mutant viral strain. Two of them had no detectable viral load at presentation. Two patients reactivated 3 and 5 months after HDT/ASCT, while 1 reactivated immediately after a bortezomib/ doxil salvage regimen. ConclusionThe risk of HBV reactivation appeared to be commonest during the immune reconstitution phase after HDT/ASCT. Although the majority of patients with HBV infection and not receiving HDT/ASCT do not reactivate, the risk may not negligible when bortezomib is used (7%). Undetectable HBV DNA and the use of anti-viral prophylaxis do not appear to preclude reactivation. The optimal use of anti-viral prophylaxis, particularly if bortezomib is given, should be further evaluated. This is particularly relevant in the current era where bortezomib plays a dominant role in the treatment of MM, and especially in endemic regions where the incidence of HBV infection is high. Disclosures:No relevant conflicts of interest to declare.

Hepatitis B virus genotypes and evolutionary profiles from blood donors from the northwest region of China

Hepatitis B virus (HBV) is prevalent in China and screening of blood donors is mandatory. Up to now, ELISA has been universally used by the China blood bank. However, this strategy has sometimes failed due to the high frequency of nucleoside acid mutations. Understanding HBV evolution and strain diversity could help devise a better screening system for blood donors. However, this kind of information in China, especially in the northwest region, is lacking. In the present study, serological markers and the HBV DNA load of 11 samples from blood donor candidates from northwest China were determined. The HBV strains were most clustered into B and C genotypes and could not be clustered into similar types from reference sequences. Subsequent testing showed liver function impairment and increasing virus load in the positive donors. This HBV evolutionary data for China will allow for better ELISA and NAT screening efficiency in the blood bank of China, especially in the northwest region.

Risk factors for paternal vertical transmission of hepatitis B virus.

To explore the risk factors of and the influence of different hepatitis B virus (HBV) DNA load on paternal vertical transmission of HBV. Totally, 161 HBsAg negative women, whose husband was HBsAg positive, attended the antenatal clinics of the Provincial Maternity and Child Health Hospital of Fujian from September 2007 to December 2008 and their newborns were selected, and the epidemiologic information, the duration of being a HBV carrier, the first class HBV family history of the fathers, HBV markers, HBV DNA load, HBsAb of the gravidas, the outcomes of the newborns were all collected. Cord blood was sampled after delivery for HBV DNA quantification and those with HBV DNA load >/= 1.0 x 10(3) copy/ml were chosen as the case group and those < 1.0 x 10(3) copy/ml as control. (1) Among the 161 newborns, 36 HBV DNA positive cord blood samples were detected, giving a rate of 22.4% (36/161) for paternal vertical transmission of HBV. The HBV DNA positive rate in cord blood was 32.0% (23/72) in HBeAg-positive fathers and 14.6% (13/89) in HBeAg-negative fathers. (2) Univariate analysis showed that HBeAg-positive, HBV DNA positive, first class family history of HBV and the duration of being a HBV carrier of the fathers were risk factors of paternal HBV vertical transmission [chi(2) = 6.892, 29.916, 29.499 and 23.821, OR = 2.7, 5.2, 8.3 and 1.4 (P < 0.01)]. (3) Multivariate analysis found that paternal serum HBV DNA positive and the first class family history of HBV of the father side were risk factors of paternal vertical transmission of HBV (OR = 11.1, 95%CI: 4.6 - 27.1; OR = 17.1, 95%CI: 3.5 - 82.6). (4) According to the different serum HBV DNA load of the HBsAg-positive father, 7 groups were divided. A dose dependent effect was found that the HBV DNA positive rate of the cord blood increased with the rising of HBV DNA load. No HBV DNA positive cord blood was detected when paternal HBV DNA load was < 1.0 x 10(4) copy/ml, while 100% of the cord blood were positive when paternal HBV DNA load >/= 1.0 x 10(8) copy/ml. (5) The average birth weight of the newborns in the two groups was the same (3.3 +/- 0.4) kg. And the delivery mode, gestational age at delivery, height and Apgar score of the newborns at 1 minute, neonatal pathological jaundice and other complications had no significant difference between the two groups (P > 0.05). No relationship was found between the neonatal outcomes and the paternal HBV vertical transmission (P > 0.05). HBV DNA load in the serum of HBsAg-positive father, and the paternal first class family history of HBV are risk factors of paternal HBV vertical transmission. When the serum HBV DNA load in HBsAg-positive father is >/= 1.0 x 10(7) copy/ml, the possibility of paternal vertical transmission of HBV would increase.

Inhibition of STAT1 methylation is involved in the resistance of hepatitis B virus to Interferon alpha

As a major therapy for hepatitis B virus (HBV) infection, Interferon alpha (IFN-alpha) triggers intracellular signal transduction including JAK-STAT pathway to produce various antiviral effector mechanisms. However, patients with chronic hepatitis B usually show low response to IFN-alpha treatment and the underlying mechanism remains unclear. In the present study, HepG2 and HepG2.2.15 cells were used to examine the Type I IFN receptors expression, phosphorylation and methylation of STAT1. STAT1–PIAS1 interaction in cells was tested by protein co-immunoprecipitation. The potential improvement of S-adenosylmethionine (SAM) in the antiviral effect of IFN-alpha was also investigated. Our data demonstrated that both chains of the Type I IFN receptors were expressed for a much higher extent in HepG2.2.15 cells than in HepG2 cells. HBV inhibited dramatically the methylation rather than the phosphorylation of STAT1, which was consistent with an increased STAT1–PIAS1 interaction. Combined with IFN-alpha, SAM treatment effectively improved STAT1 methylation and attenuated STAT1–PIAS1 binding, followed by increased PKR and 2′,5′-OAS mRNA expression, thus significantly reducing the HBsAg, HBeAg protein levels and HBV DNA load in the supernatant of HepG2.2.15 cells. Less STAT1 methylation and subsequent increased STAT1–PIAS1 interaction are involved in the mechanism of the IFN-alpha-antagonistic activity of HBV. By improving STAT1 methylation, SAM can enhance the antiviral effect of IFN-alpha.

Hepatitis B Virus Load in Serum Does Not Reflect Histologic Activity in Patients With Decompensated Cirrhosis

Little is known about whether histologic data can predict which patients with hepatitis B virus (HBV)-related decompensated cirrhosis will respond to antiviral therapies. We assessed the relationship between serum HBV DNA load and histologic activity by analyzing liver specimens from patients with decompensated cirrhosis. The study included 72 consecutive patients who underwent liver transplantation for HBV-related decompensated cirrhosis between November 2000 and March 2008. None of the patients had received nucleoside or nucleotide analogues more than 2 weeks before transplantation. Serum HBV DNA levels at the time of transplantation were compared with histologic activity in explanted liver specimens. The median HBV DNA level of the 72 patients was 5.40 log(10) copies/mL (range, 1.45-8.00 log(10) copies/mL). There were no differences in HBV DNA level between patients grouped according to lobular or portoperiportal activity (P = .678, P = .291, respectively). Of 16 patients (22.2%) with HBV DNA levels less than 2000 copies/mL, 8 patients (50.0%) had moderate or severe portoperiportal activity; their median alanine aminotransferase level was 30.5 U/L (range, 12-135 U/L). HBV DNA load does not reflect histologic activity in patients with HBV-related decompensated cirrhosis. Although patients with decompensated cirrhosis might have normal levels of alanine aminotransferase and a low level of viremia (<2000 copies/mL), they still can have significant portoperiportal activity.

Molecular and serological characterization of hepatitis B virus genotype A and D infected blood donors in Poland

Hepatitis B virus (HBV) genotypes have distinct geographical distributions and influence severity of clinical outcome and response to antiviral therapies. HBV polymorphism in HBV surface antigen (HBsAg) positive first time blood donors from Poland was examined. HBV serological markers and HBV DNA were tested in 170 samples. Whole genome (n = 53) or specific region sequences: pre-S/S and basic core promoter/precore (BCP/PC) region (91 and 154 samples, respectively) were phylogenetically analyzed. The median age of infected donors was 21 years. Anti-HBs, anti-HBe and hepatitis B e antigen were detected in 5%, 92.4% and 10.5% of tested donors, respectively. The HBV DNA load ranged between unquantifiable and 3.1 x 10(10) IU/mL (median: 4.10 x 10(3) IU/mL). Genotypes A2 (81.2%) and D (18.8%) co-circulated. Phylogenetic analyses revealed differences between the genotypes. Viral load and level of HBsAg tended to be lower in genotype D. The median HBsAg/HBV DNA ratio expressed in IU/mL was one for both genotypes, but very low or very high ratios appeared more frequent in genotype D infections. Higher amino acid variability in the surface proteins (median: 4%vs 1.5%; P = 0.01) and in the major hydrophilic region was observed in genotype D (P = 0.01). BCP/PC region analysis revealed the double mutation 1762T/1764A in 49/125 (39.2%) genotype A2 and 6/29 (20.7%) genotype D strains (P = 0.08). Mutations in PC and BCP regions correlated neither with HBsAg nor HBV DNA levels. HBV genotype A2 is dominant in HBsAg positive donors in Poland. Minority genotype D strains are significantly more substituted than genotype A2 strains potentially affecting the course of infection.

Prevalence of human cytomegalovirus co-infection in patients with chronic viral hepatitis B and C: A comparison of clinical and histological aspects

BackgroundHuman cytomegalovirus (HCMV) is a common pathogen of severe disease in patients with impaired immune functions. Reactivation of HCMV in immunocompetent host is usually asymptomatic, but may deteriorate the prognosis of patient with chronic illness. ObjectivesThis study was conducted to detect HCMV infection in patients with chronic hepatitis B virus (HBV) and chronic hepatitis C virus (HCV) infections and to point out the effects of HCMV-HBV and HCMV-HCV co-infections on liver histology. Study designExpression of HCMV DNA was determined in liver tissue biopsies by real-time quantitative polymerase chain reaction (qPCR) method. There were 44 chronic HBV, and 25 chronic HCV patients within the study group. Control group consisted of 36 patients with hepatologic malignancies. ResultsHCMV infection was demonstrated in 52.3% of chronic HBV, and 36% of chronic HCV patients. Although alanine aminotransferase (ALT) levels of HCMV-infected HBV patients were decreased slightly, they were increased in HCV patients. Histologic activity scores (necroinflammation and fibrosis) of HCMV-positive patients were higher than that of HCMV-negatives in both HBV and HCV groups. Intrahepatic HBV DNA or HCV RNA loads of the corresponding study groups were decreased in HCMV-infected patients. ConclusionWe conclude that HCMV infection is common in chronic HBV and HCV patients, who can be regarded as patients at high risk for HCMV disease. Though the histological changes were more marked in liver, replication of HBV and HCV were inhibited in HCMV-positive cases.

Clinical efficacy of adefovir dipivoxil combined with anti-tuberculosis drugs inn treatment for chronic hepatitis B complicated with pulmonary tuberculosis

Objective It is purposed to evaluate clinical efficacy of adefovir dipivoxil combined with anti-tuberculosis (TB) drugs in treatment for chronic hepatitis B (HB) complicated with pulmonary TB. Methods Ninety-four eases of pulmonary TB complicated with chronic HB receiving anti-TB drug treatment at Liaocheng Hospital for Infectious Disease during September 2006 to September 2008 were divided into treatment and control groups matched on their pulmonary TB lesion, damage in liver function and HB virus (HBV) DNA load. Forty-seven cases in the treatment group were administered with anti-TB drugs, i.e., isoniazid (INH) 0. 3 g daily, rifapentine (RFT) 0.45 g twice a week, pyrazinamide (PZA) 1.5 g daily, streptomycin (SM) 0. 75 g daily or ethambutol (EMB) 0. 75 g daily, and regular liver-protection drags, as well as oral adefovir dipivoxil 10 mg daily for eight months, and another 47 cases in the control group were administered with the same anti-TB drugs and regular liver-protection drugs as the treatment group. Two to eight weeks after treatment, clinical symptoms, liver function, HBV DNA load, negative conversion of acid-fast staining bacteria in sputum smear, and absorption of pulmonary TB lesion were evaluated for all the patients in both groups. Data were analyzed with U-test and χ2-test, respectively. Results After treatment, damage in liver function worsened in three cases (6.4%), as compared to that in 47.3 percent of the control group (P<0.05). Clinical symptoms, including tiredness, loss of appetite, abdominal distension, uncomfortable in upper abdomen, and so on, appeared in three cases of the treatment group (6.4%) during anti-TB treatment, as compared to those in 18 cases of the control group (47.3%) (P<0.05). HBV DNA conversed to negative (less than 1.0×103 lg copies/ml) in 24 cases of the treatment group (51.1%), HBV load went down to 1.0×103-1.0×104 lg copies/ml in 23 cases (48. 9%), sputum smear bacteria conversed to negative in 41 cases (87. 2%) and pulmonary lesion absorbed in 45 cases (95.7%), all with statistically significance as compared to those in the control group (47. 3% , 0. 0% , 2. 6% , 39. 5% and 50. 0% , respectively, P <0. 05). Renal function was all in normal in both groups and no patient with drug resistant variant HBV was found during and after treatment, and no obviously adverse reaction was found in patients with adefovir dipivoxil. Conclusion Adefovir dipivoxil has a good safety and can reduce laver damage in anfi-TB drug treatment for patients of chronic hepatitis B complicated with pulmonary TB. Key words: Hepatitis B, chronic; Tuberculosis,pulmonary; Antitubercular agents

Viral suppression correlates with dendritic cell restoration in chronic hepatitis B patients with autologous cytokine‐induced killer cell transfusion

Myeloid and plasmacytoid dendritic cells (mDCs, pDCs) are functionally impaired in patients with chronic hepatitis B (CHB). Adoptive immunotherapy can suppress hepatitis B virus (HBV) replication in CHB patients, but whether it can restore the functionality of mDCs and pDCs remains unknown. Autologous cytokine-induced killer (CIK) cells obtained from 14 CHB patients were transfused back to patients, case by case, to observe the effect of CIK-cell treatment on the frequency and functionality of mDCs and pDCs in CHB patients during a 24-week follow-up investigation. Seven virological responders exhibited a sustained decrease in HBV load after CIK-cell transfusion; another seven non-virological responders showed only sustained high levels of HBV load during the 24-week period following CIK-cell transfusion. The rate of hepatitis B e antigen loss or seroconversion was also higher in virological responders than in non-virological responders. Importantly, we found that the frequency and cytokine-producing capacity of mDCs and pDCs increased significantly in virological responders, but not in non-virological responders. In addition, these patients exhibited a close correlation between restoration DC subsets and a decrease in HBV DNA load, rather than a change in the alanine aminotransferase level. Cytokine-induced killer-cell treatment reduced HBV DNA load in some CHB patients; the efficiency at least partially correlates with the restoration of frequency and functionality of mDCs and pDCs.

Peripheral T-lymphocyte subpopulations in different clinical stages of chronic HBV infection correlate with HBV load.

To characterize the peripheral T-cell subpopulation profiles and their correlation with hepatitis B virus (HBV) replication in different clinical stages of chronic HBV infection. A total of 422 patients with chronic HBV infection were enrolled in this study. The patients were divided into three stages: immune-tolerant stage, immune active stage, and immune-inactive carrier stage. Composition of peripheral T-cell subpopulations was determined by flow cytometry. HBV markers were detected by enzyme-linked immunosorbent assay. Serum HBV DNA load was assessed by quantitative real-time polymerase chain reaction. CD8(+) T-cells were significantly higher in patients at the immune-tolerant stage than in patients at the immune-active and -inactive carrier stages (36.87 +/- 7.58 vs 34.37 +/- 9.07, 36.87 +/- 7.58 vs 28.09 +/- 5.64, P < 0.001). The peripheral blood in patients at the immune-tolerant and immune active stages contained more CD8(+) T-cells than CD4(+) T-cells (36.87 +/- 7.58 vs 30.23 +/- 6.35, 34.37 +/- 9.07 vs 30.92 +/- 7.40, P < 0.01), whereas the peripheral blood in patients at the immune-inactive carrier stage and in normal controls contained less CD8(+) T-cells than CD4(+) T-cells (28.09 +/- 5.64 vs 36.85 +/- 6.06, 24.02 +/- 4.35 vs 38.94 +/- 3.39, P < 0.01). ANOVA linear trend test showed that CD8(+) T-cells were significantly increased in patients with a high viral load (39.41 +/- 7.36, 33.83 +/- 7.50, 31.81 +/- 5.95 and 26.89 +/- 5.71, P < 0.001), while CD4(+) T-cells were significantly increased in patients with a low HBV DNA load (37.45 +/- 6.14, 33.33 +/- 5.61, 31.58 +/- 6.99 and 27.56 +/- 5.49, P < 0.001). Multiple regression analysis displayed that log copies of HBV DNA still maintained its highly significant coefficients for T-cell subpopulations, and was the strongest predictors for variations in CD3(+), CD4(+) and CD8(+) cells and CD4(+)/CD8(+) ratio after adjustment for age at HBV-infection, maternal HBV-infection status, presence of hepatitis B e antigen and HBV mutation. Differences in peripheral T-cell subpopulation profiles can be found in different clinical stages of chronic HBV infection. T-cell impairment is significantly associated with HBV load.

Symptomatic Hepatitis B Virus (HBV) Reactivation despite Reduced Viral Fitness Is Associated with HBV Test and Immune Escape Mutations in an HIV‐Coinfected Patient

Two sequential hepatitis B virus (HBV) strains obtained before and during an icteric flare-up of an occult HBV infection in a patient coinfected with human immunodeficiency virus revealed HBV surface antigen (HBsAg) test escape mutations, although the patient had never received hepatitis B-specific immunoglobulin. In contrast to the high HBV DNA loads, recurrence of HBsAg, and resulting icteric hepatitis, phenotypic analysis of the mutated HBV strains revealed significantly reduced replication efficacies in vitro, compared with wild-type HBV. Therefore, immune escape in the transiently anti-HBs-positive patient appeared to be crucial for persistence and reactivation. Immune escape mutants evolved even without exogenous selective pressure, hampered detection in HBsAg screening, and might be transmitted during reactivation with high HBV loads.

A study on the relationship between point mutation in pre-core region G1896A of hepatitis B virus and safety of breast feeding

To investigate the relationship between pre-core G1896A point mutation of hepatitis B virus (HBV) and safety of breast feeding. Serum and breast milk samples were collected from 62 pregnant women of HBV DNA positive/HBeAg negative. PCR-solid phase hybridization was used to detect the point mutation in pre-core region G1896A of HBV from pregnant women, and HBV DNA loads in sera and breast milk were determined by fluorescence quantitative PCR (FQ-PCR). The prevalence of point mutation was 61.3% (38/62) in 62 pregnant women with HBsAg positive/HBeAg negative. The positive rate of HBV DNA in breast milk of group with point mutation (28.9%) was similar to that of group without mutation (29.2%, chi2=0.0003, P>0.05). However, The positive rate of HBV DNA in breast milk of group with high HBV loads (56.0%) was significantly higher than that of group with low HBV loads (10.8%, chi2=14.79, P<0.01). The point mutation in pre-core region G1896A of HBV dose not affect the positive rate of HBV DNA in breast milk and higher HBV DNA loads in serum of pregnant women might increase the risk of mother-infant transmission.

A new method of concentrating hepatitis B virus (HBV) DNA and HBV surface antigen: an application of the method to the detection of occult HBV infection

The risk of post-transfusion hepatitis B virus (HBV) infection has been reduced after the implementation of HBV nucleic acid amplification technology (NAT). However, the problem of HBV DNA-positive and HBV surface antigen (HBsAg)-negative occult HBV infections remains to be solved. This is in part due to the HBV DNA load being too low to detect these occult HBV infections using mini-pool NAT. In Japan, the assay for the antibody against the HBV core antigen (anti-HBc) has not completely excluded occult HBV infection. To solve this problem, we have developed a new method of concentrating HBV DNA and HBsAg simultaneously to increase the sensitivity of detection tests. Virus concentration is achieved by the enhancement of the agglutination of viruses using poly-L-lysine in the presence of a bivalent metal. Poly-L-lysine-coated magnetic beads are used to shorten the time of each step of the concentration procedure. Seventy-seven anti-HBc-positive and HBsAg-negative donations were examined. HBsAg and anti-HBc were tested by enzyme immunoassay (EIA) (AxSYM; Abbott) and haemagglutination inhibition test (Japanese Red Cross), respectively. HBV surface antigen and HBV DNA levels were concentrated up to four- to sevenfold. Using this method, 35 of the 77 anti-HBc-positive and HBsAg-negative donors were HBV DNA-positive by individual NAT and a further five donors became HBV DNA-positive by HBV concentration. Twenty-seven of 40 occult HBV infections became HBsAg-positive by HBsAg concentration. Our new method of concentrating HBV and HBsAg increased the sensitivities of EIA and HBV NAT, and enabled us to detect 27 of 40 occult HBV infections by HBsAg EIA.

DIAGNOSING DIFFERENT STAGES OF HEPATITIS B INFECTION USING A COMPETITIVE POLYMERASE CHAIN REACTION ASSAY

Purpose: Different stages of hepatitis B virus (HBV) infection can be defined by serum HBV DNA levels. This study attempts to (1) investigate serum HBV DNA levels in inactive carriers and patients with chronic HBV (CHB) infection and (2) define cut-off value between inactive carriers and HBeAg (precore antigen of HBV) negative CHB patients in Indian population. Methods: One hundred and forty samples encompassing 42 inactive HBsAg carriers and 98 CHB patients (53 HBeAg-positive and 45 HBeAg-negative) were analysed. Serum HBV DNA levels were determined employing an in-house competitive polymerase chain reaction (cPCR) assay. Results: The HBeAg-positive patients were found to have the maximum median HBV DNA load, which was significantly higher than the HBeAg-negative ones (median; 1.25 ×108vs. 2.30 ×105copies/mL; P < 0.05). Interestingly, the latter group has significantly higher HBV DNA levels than the inactive carriers (median; 2.30 ×105vs. 4.28 ×103copies/mL; P < 0.05). The 2.5 ×104copies/ml HBV DNA levels were optimal for discriminating CHB patients (HBeAg-negative) from inactive carriers with 75.6 and 78.6% sensitivity and specificity, respectively. Conclusions: Despite the extensive overlapping of HBV DNA levels in inactive carriers and HBeAg negative CHB patients, 2.5 ×104copies/mL is the most favourable cut-off value to classify these individuals and would be imperative in the better management of this dreadful disease.