Hepatitis B Virus DNA Load Research Articles

HBV Vaccine Escape Mutations in a Chronic Hepatitis B Patient Treated with Nucleos(t)ide Analogues

The hepatitis B virus (HBV) polymerase (pol) gene completely overlaps with the envelope (S) gene. Nucleos(t)ide analogue (NA) resistance mutations in the pol gene of HBV, either from selection of primary or secondary resistance mutations, typically result in changes in the overlapping hepatitis B surface antigen (HBsAg). Recent studies have conferred a new acronym to these HBV pol/S gene overlap mutants; ADAPVEMs, for antiviral drug-associated potential vaccine-escape mutants. The present report aimed to assess the determined multiple HBV vaccine-escape mutants in a Turkish patient with chronic hepatitis B (CHB), undergoing NAs treatment. The liver biopsy of HBsAg positive, HBeAg negative 53-year old female patient with CHB, revealed a score as histological activity index; 9 and fibrosis; 2 according to Ishak classification. NA treatment backgrounds consisted of 24 months lamivudine, followed by 18 months entacavir and lastly 3 months tenofovir monotherapies. Since HBV DNA load was determined as 7.030.000 IU/ml at the 4th month of tenofovir therapy, entecavir was added as current treatment regimen, and tenofovir + entecavir therapy decreased the HBV DNA load (400 IU/ml). Sequence analysis was performed for HBV pol/S gene and overlapping pol/S gene amino acid substitutions, primary/compensatory NA resistance mutations and antiviral drug-associated potential vaccine-escape mutations (ADAPVEM) were analysed. The patient isolate was identified as genotype D/subgenotype D1 of HBV. Primary drug resistance mutations (rtV173L + rtL180M + rtM204V) to lamivudine and telbivudine and a compensatory mutation (rtQ215H) to lamivudine and adefovir were described in the HBV pol gene sequence. However, multiple HBV vaccine-escape mutations (sS143T + sD144E + sG145R + sE164D + sI195M) have been determined on the HBV overlapping pol/S gene region. Lamivudine and telbivudine which are the frequently preferred drugs for the treatment of CHB in Turkey, have the potential to lead to ADAPVEMs. Thus ADAPVEMs should be monitored in infected and NA treated CHB patients and their public health risks should be assessed.

Entecavir and hepatitis B immune globulin in patients undergoing liver transplantation for chronic hepatitis B

For patients undergoing liver transplantation (LT) for hepatitis B virus (HBV)–related liver disease, the current standard of care for preventing reinfection of the allograft is nucleoside analogue therapy combined with hepatitis B immune globulin (HBIG). Entecavir has demonstrated high efficacy and a favorable safety profile for chronic hepatitis B (CHB) treatment, but data for patients undergoing HBV-related LT are limited. This study assessed the safety and efficacy of entecavir combined with various HBIG regimens after CHB-related LT. In this phase 3b, single-arm, open-label study, 65 patients undergoing LT for CHB-related liver disease with an HBV DNA load <172 IU/mL at LT received entecavir (1.0 mg daily) for 72 weeks after LT. The primary endpoint was the proportion of evaluable patients (treated for ≥4 weeks) with virological recurrence (HBV DNA level ≥50 IU/mL) through week 72. Concomitant HBIG therapy was received by 64 of the 65 enrolled patients, and 44% of these patients received high-dose HBIG (any HBIG dose in the specified interval ≥10,000 IU). Through week 72, all 61 patients evaluable for the efficacy analysis had undetectable HBV DNA. The Kaplan-Meier estimate of patients without hepatitis B surface antigen (HBsAg) recurrence at week 72 was 0.9655. Two patients experienced a reappearance of HBsAg, but both remained HBV DNA− until the last follow-up. The frequency and nature of adverse events were consistent with those expected for this patient population. Serum creatinine increments ≥0.3 mg/dL and ≥0.5 mg/dL occurred in 62% and 39% of the patients, respectively, and all of these patients received calcineurin inhibitor therapy. In conclusion, in this population of patients treated with entecavir after CHB-related LT, entecavir was well tolerated and effective in maintaining viral suppression, even in individuals who experienced a reappearance of HBsAg. Liver Transpl 19:887–895, 2013. © 2013 AASLD.

SAT0079 Analysis of Rheumatoid Factor According to Various Hepatitis B Virus Infectious Statuses and Vaccination

BackgroundRheumatoid factor (RF) is produced as a result of polyclonal B cell activation, but the reasons for its production are still unknown. RF positivity can be seen in several diseases...

Correlation of hepatitis B virus X antigen expression with liver cell apoptosis

To study the roles of hepatitis B virus (HBV) X antigen (HBxAg) in development of HBV-related liver diseases and carcinogenesis. Liver tissues were collected from patients with HBV infection (HBV carriers, n = 14; chronic hepatitis B (CHB), n = 24), HBV-related liver cirrhosis (LC, n = 20), or hepatocellular carcinoma (HCC, n = 20). Immunohistochemistry was used to detect the expression of HBxAg and the host apoptosis-related genes Fas and Fas ligand (Fas-L). The correlations of HBxAg with HBV DNA level in serum, inflammation grade, and fibrosis stage were statistically analyzed. Liver inflammation grade and fibrosis stage were in accordance with Knodell standard. x2test and Fisher's exact test were adopted in count data, x2split method was adopted in pariwise comparisons between multiple samples, Rank-sum test was adopted in ranked data, Spearman rank correlation analysis was adopted in correlation analysis. The rates of HBxAg-positivity were similar between the patients with HBV infection (71.1%), LC (60.0%), and HCC (65.0%) (x2= 0.754, P = 0.686). The rates of Fas- and Fas-L-positivity in liver cells were also similar between the three groups (Fas: 28.9% vs. 20.0% vs. 5.0%, x2= 4.667, P = 0.101; Fas-L: 36.8% vs. 50.0% vs. 60.0%, x2= 2.988, P = 0.225). However, the positive rate of Fas in lymphocytes of liver tissue was significantly higher in the HCC patients than in the HBV-infected patients (90.0% vs. 68.4%, Z = -4.360, P = 0.00001). The expressions of HBxAg and Fas-L corresponded to regions of severe inflammation in tissues from LC patients and some HCC patients. Furthermore, the expression of HBxAg was positively correlated with Fas (r = 0.304, P = 0.02) and Fas-L (r = 0.368, P = 0.004) in the HBV-infected patients and LC patients, and the expression of Fas was positively correlated with that of Fas-L (r = 0.448, P = 0.0004). Patients with high and medium loads of HBV DNA showed significantly higher rates of HBxAg-positivity than those with low loads (88.9% and 69.2% vs. 26.7%, P less than 0.05). In the early stage of chronic HBV infection, HBxAg may induce liver cell apoptosis by up-regulating Fas expression, and in the later stage, HBxAg may induce immune escape by up-regulating Fas-L expression in liver cells. Together, HBxAg and high HBV DNA load may promote chronic HBV infection and progression to hepatocarcinogenesis.

A pregnant woman with acute hepatitis B in whom vertical transmission was prevented by tenofovir disoproxil fumarate.

The optimal management of acute hepatitis B in pregnant women remains to be fully evaluated. A case of hepatitis B virus (HBV) infection in a 20-year-old pregnant woman who initially presented with jaundice is reported. Serum samples were positive for anti-HBc IgM and HBe antigen. Her husband was infected with HBV genotype A and received entecavir because of prolonged hepatitis and a high viral load. The HBV DNA sequence of the wife completely matched that of her husband, indicating that he was the source of HBV infection. In accordance with guidelines for the treatment of chronic HBV carriers, the wife started to receive tenofovir disoproxil fumarate (TDF) in her third trimester. After 4months of treatment, the HBV DNA load decreased from 7.6 to 3.5log copies/ml. At delivery, the serum was found negative for the HBe antigen. Seven months after treatment began, the HBs antigens also disappeared. The baby, totally healthy, received passive-active immunoprophylaxis. At 3months, the baby remained free of HBV infection. TDF thus prevented exacerbation and prolongation of acute HBV infection in a pregnant woman. Subsequent treatment also prevented mother-to-infant transmission of HBV. The clinical course of her husband, who had HBV infection and received entecavir, is also reported.

Epidemiological and clinical features of occult hepatitis B in HIV infection without antiretroviral treatment

To investigate and analyze the differential prevalence, as well as the risk factors and clinical features, of occult hepatitis B virus (HBV) infection in the human immunodeficiency virus (HIV)-infected population without antiretroviral therapy (ART) as compared to the general (non-HIV-infected) population. Two-hundred-and-forty-eight individuals with confirmed HIV infection but ART naive (males: 220, females: 28; 15-82 years old) were enrolled in the study, along with 121 healthy individuals (confirmed HIV antibody-negative; males: 53, females: 68; 20-88 years old). HBV markers (hepatitis B surface antigen (HBsAg); hepatitis B e antigen (HBeAg); anti-HBs, anti-HBe and anti-hepatitis B core (HBc) antibodies) were detected by microparticle enzyme-linked immunosorbent assay (AxSYM immunology analyzer manufactured by Abbott Laboratories); all cases and controls were confirmed negative for hepatitis B surface antigen (HBsAg). Then, the HBV DNA level in serum was detected using nucleic acid amplification assay (COBAS AmpliPrep/COBAS TaqMan HBV test, version 2.0 manufactured by Roche). CD4+ T lymphocytes were measured by flow cytometry, and alanine aminotransferase (ALT, marker of liver function) was measured by enzymatic assay. Twenty-four of the HIV cases (9.7%) and four of the healthy controls (3.3%) tested positive for HBV DNA; the amount of individuals with HBV DNA-positivity was significantly higher in the HIV-infected group (P = 0.035). Among the 24 cases of HBV DNA(+) HIV-infected individuals, the lowest HBV DNA load was < 20 IU/ml and the highest was 3.22 x 10s IU/ml; nine of the individuals (37.5%) had HBV DNA load > 100 IU/ml, four (16.7%) had 20-99 IU/ ml, and 11 (45.8%) had < 20 IU/ml. Among the total HIV-infected cases with HBV DNA-positivity, 7.3% (8/110) were anti-HBc(+)/anti-HBs(+), 20.8% (11/53) were anti-HBc(+)/anti-HBs(-), 14.3% (3/21) were anti-HBc(-)/anti-HBs(+), and 3.1% (2/64) were anti-HBc(-)/anti-HBs(-). The amount of individuals with HBV DNA-positivity in the anti-HBc(+)/anti-HBs(-) group was significantly different from those in the anti-HBc(+)/anti-HBs(+) group (P = 0.018) and the anti-HBc(-)/anti-HBs(-) group (P = 0.003). However, multiple comparison of HBV DNA loads detected between the four groups of HBV marker status revealed no significant difference (P = 0.805). Furthermore, statistical analysis provided no evidence to support that occult hepatitis B infection in HIV-infected individuals had any impact on CD4+ T lymphocytes count (Z = 1.902, P = 0.059) or ALT levels (Z =1.401, P = 0.161). HIV-infected individuals who are ART naive and HBsAg(-) have a higher incidence of HBV DNA-positivity than individuals in the general (non-HIV-infected) population. In addition, the highest rate of occult hepatitis B among the HIV-infected cases occurred among individuals who were anti-HBc(+)/anti-HBs(-).

Downregulation and altered function of natural killer cells in hepatitis B virus patients treated with entecavir

The aim of the present study was to investigate the natural killer (NK) cell phenotype and function in chronic hepatitis B virus (HBV) patients and to study the effects of entecavir therapy (10 mg/day, p.o.) on these responses. Peripheral blood NK cells were collected from 18 chronic HBV patients and 14 healthy controls. The effect of entecavir therapy on the phenotype and function of NK cells in chronic HBV patients was characterized by flow cytometry analysis. Concentrations of alanine aminotransferase (ALT) and aspartate aminotransferase (AST), HBV viral loads in both groups and potential associations between the frequency of peripheral NK cell subsets and clinical measures were determined. There was a significant reduction in the number of CD3(-)CD56(+) NK cells in chronic HBV patients compared with healthy controls. Furthermore, there were significant increases in the percentage of CD3(-)CD56(+)NKG2D(+) and CD3(-)CD56(+)NKP30(+) NK activating receptors in chronic HBV patients compared with healthy individuals, who exhibited downregulated expression following entecavir treatment. Spearman's correlation analysis revealed that there was a significant positive correlation between the percentage of NKG2D(+) and NKP30(+) NK cells and serum ALT levels. Characterization of NK cell degranulation indicated that the frequency of CD107a(+) NK cells in HBV patients (in response to K562 stimulation) was significantly greater than in healthy controls but decreased following entecavir treatment. Entecavir treatment of hepatitis B e antigen-positive chronic HBV-infected patients not only led to a reduction in HBV DNA loads and normalization of ALT and AST levels, but also resulted in the recovery of NK cell-mediated immunity.

The correlation between T helper type 17 cells and clinical characters in Chinese paediatric patients with chronic hepatitis B

Interleukin (IL)-17-mediated immune response has been shown to play a critical role in inflammation-associated disease. However, its role in the pathogenesis of chronic hepatitis B virus (HBV) in paediatric patients remains unknown. We investigated the frequency of T helper type 17 (Th17) cells and evaluated the association between the Th17 and clinical characters in paediatric patients with chronic hepatitis B (CHB). The frequency of Th17 cells was detected by flow cytometry analyses from 65 paediatric patients with CHB and nine healthy controls. The degree of hepatic inflammation was graded using the histological activity index (HAI). Compared with healthy controls, the frequency of Th17 cells in peripheral blood was significantly higher in paediatric patients with CHB. The proportion of Th17 cells was higher in the patients with higher HAI score (G2-G3) compared to those subjects with lower HAI score (G0-G1), but the frequency of Th17 cells had no correlation with serum HBV DNA loads or alanine aminotransferase levels. Compared with the younger age group (age 1-6 years), Th17 cell frequency was higher in the older age group (age 7-18 years). Peripheral Th17 cell frequency is associated closely with inflammation activity of liver tissues in paediatric patients with CHB.

Reactivation of occult hepatitis B virus infection following cytotoxic lymphoma therapy in an anti‐HBc negative patient

Screening hepatitis B virus (HBV) surface antigen (HBsAg) and HBV core antibody (anti-HBc) is recommended prior to cytotoxic or immunosuppressive therapy. This case describes an anti-HBc negative, DNA positive occult HBV infection in a 71-year-old Caucasian male following rituximab-based treatment for follicular lymphoma. Pre-screening serology indicated negative HBsAg and anti-HBc. However, following sequential treatment cycles the patient developed weak HBsAg with a low HBV DNA load (<1,000 IU/ml), but remained anti-HBc negative. The DNA load peaked 5 months later (>1 × 10(6) IU/ml) and he was subsequently treated with Tenofovir. Currently the patient remains anti-HBc negative, and is anti-HBe negative, anti-HBs negative, HBeAg positive. No clinical or biochemical evidence of hepatitis has occurred. Sequencing and phylogenetic analysis identified the HBV genosubtype as D4, most probably acquired some years ago during a stay in Papua New Guinea, in spite of prior hepatitis B vaccination. Four amino acid substitutions were detected within the HBsAg loop yet none in the core protein. This case questions the dependability of anti-HBc testing and highlights the role of HBV DNA testing prior to and throughout cytotoxic or immunosuppressive regimes. As this case exemplifies, vaccination protects against clinical infection but may not exclude seronegative occult infection with the possibility of reactivation.

Long-Term Follow-up of Children With Postnatal Immunoprophylaxis Failure Who Were Infected With Hepatitis B Virus Surface Antigen Gene Mutant

The long-term evolution and outcomes of infection with a hepatitis B virus (HBV) surface antigen (HBsAg) gene mutant (hereafter, "HBsAg-mutant HBV") among immunized children remain unclear. Ninety-five HBsAg-positive children aged 0.5-18.4 years (mean age, 5.8 years) who did not respond to postnatal immunoprophylaxis were prospectively followed for 1-22.8 years (mean follow-up, 8.5 years). Clinical, serologic, and virologic features were compared between 38 untreated HBV e antigen (HBeAg)-positive children carrying HBsAg-mutant HBV (group 1) and 49 children carrying wild-type HBV (group 2). HBsAg-mutant HBV was examined in 20 immunized children presenting with HBV-related hepatocellular carcinoma (HCC). The initial alanine aminotransferase (ALT) level, the maximal ALT level during the HBeAg-positive phase of HBV infection, the cumulative incidence of HBeAg seroconversion (P = .0018), and the rate of low serum HBV DNA load (defined as <10 copies/mL) at the last visit (P = .006) were higher in group 1 than in group 2. A higher frequency of HBV genotype C and a higher ALT level during surface mutant viremia were observed in codon 110-129 mutants than in codon 144-145 mutants. None of the 95 patients developed cirrhosis or HCC. HBsAg-mutant HBV was detected in 3 of 8 (38%) HBV DNA-positive children with HCC. HBeAg-positive immunized children carrying HBsAg-mutant HBV may develop hepatitis activity, HBeAg seroconversion, and a low viremic state earlier than those carrying wild-type HBV. Continuous monitoring of children with wild-type HBV infection and those with HBsAg-mutant HBV for possible development of HCC is needed.

De novocombined lamivudine and adefovir dipivoxil therapyvsentecavir monotherapy for hepatitis B virus-related decompensated cirrhosis

To compare efficacy of combined lamivudine (LAM) and adefovir dipivoxil (ADV) therapy with that of entecavir (ETV) monotherapy for hepatitis B virus (HBV)-related decompensated liver cirrhosis. A total of 120 naïve patients with HBV-related decompensated cirrhosis participated in this study. Sixty patients were treated with combined LAM and ADV therapy (LAM + ADV group), while the other 60 were treated with ETV monotherapy (ETV group) for two years. Tests for liver and kidney function, alpha-fetoprotein, HBV serum markers, HBV DNA load, prothrombin time (PT), and ultrasonography or computed tomography scan of the liver were performed every 1 to 3 mo. Repeated measure ANOVA and the χ(2) test were performed to compare the efficacy, side effects, and the cumulative survival rates at 48 and 96 wk. Forty-five patients in each group were observed for 96 wk. No significant differences in HBV DNA negative rates and alanine aminotransferase (ALT) normalization rates at weeks 48 (χ(2) = 2.12 and 2.88) and 96 (χ(2) = 3.21 and 3.24) between the two groups were observed. Hepatitis B e antigen seroconversion rate in the LAM + ADV group at week 96 was significantly higher in the ETV group (43.5% vs 36.4%, χ(2) = 4.09, P < 0.05). Viral breakthrough occurred in 2 cases (4.4%) by week 48 and in 3 cases (6.7%) by week 96 in the LAM + ADV group, and no viral mutation was detected. In the ETV group, viral breakthrough occurred in 1 case (2.2%) at the end of week 96. An increase in albumin (F = 18.9 and 17.3), decrease in total bilirubin and in ALT (F = 16.5, 17.1 and 23.7, 24.8), reduced PT (F = 22.7 and 24.5), and improved Child-Turcotte-Pugh and the model for end-stage liver disease scores (F = 18.5, 17.8, and 24.2, 23.8) were observed in both groups. The cumulative rates of mortality and liver transplantation were 16.7% (10/60) and 18.3% (11/60) in the LAM + ADV and ETV groups, respectively. Both LAM + ADV combination therapy and ETV monotherapy can effectively inhibit HBV replication, improve liver function, and decrease mortality.

Correlation between Hepatitis B surface antigen titers and HBV DNA levels

Background/Aim:To assess the correlation between serum HBsAg titers and hepatitis B virus (HBV) DNA levels in patients with hepatitis B envelop antigen-negative (HBeAg −ve) HBV genotype-D (HBV/D) infection.Patients and Methods:A total of 106 treatment- naïve, HBeAg −ve HBV/D patients were included; 78 in the inactive carrier (IC) state and 28 in the active hepatitis (AH) stage. HBV DNA load and HBsAg titers were tested using TaqMan real-time polymerase chain reaction (PCR) and automated chemiluminescent microparticle immunoassay, respectively.Results:The median (range) log10 HBsAg titer was significantly lower in the IC group compared with AH group, 3.09 (−1 to –4.4) versus 3.68 (−0.77 to 5.09) IU/mL, respectively; P < 0.001. The suggested cutoff value of HBsAg titer to differentiate between the two groups was 3.79 log10 IU/mL. In addition, there was a significant positive correlation between HBsAg and HBV DNA levels in the whole cohort, AH, and IC groups (r = 0.6, P < 0.0001; r = 0.591, P = 0.001; and r = 0.243, P = 0.032, respectively).Conclusion:Serum HBsAg titers may correlate with HBV DNA in treatment-naïve HBeAg –ve HBV/D patients, and supports the use of HBsAg levels in clinical practice as a predictor of serum HBV DNA levels.

Anti-viral therapy to reduce recurrence and improve survival in hepatitis B virus-related hepatocellular carcinoma

Hepatocellular carcinoma (HCC) is the most common malignancy and the third leading cause of cancer death worldwide. Chronic infection with hepatitis B virus (HBV) and hepatitis C virus accounts for approximately 75%-80% of HCC cases worldwide. In particular, chronic HBV infection is a predominant risk factor for HCC in Asia and Africa. Hepatic resection and radiofrequency ablation are increasingly used for the curative treatment of HCC, and good local control can be achieved. However, the high rate of recurrence is a major obstacle to improving prognosis. A high viral load of HBV DNA is the most important correctable risk factor for recurrence. Furthermore, interferon and/or nucleotide analogues may decrease HBV DNA. Therefore, these drugs may decrease recurrence. In this article, treatment strategies for HBV-related HCC are described in order to reduce recurrence and improve survival.

Treatment with telbivudine positively regulates antiviral immune profiles in Chinese patients with chronic hepatitis B.

Chronic infection with hepatitis B virus (HBV) is associated with impairment of T and NK cell immunity. This study was aimed at investigating the impact of treatment with telbivudine (LDT) on T and NK cell immunity in patients with chronic hepatitis B (CHB). A total of 54 CHB patients and 30 healthy controls (HC) were recruited. Individual patients were treated orally with 600 mg LDT daily for 13 months. The serum HBV DNA loads, the levels of the HBV-related biomarkers alanine aminotransferase (ALT) and aspartate transaminase (AST), and the numbers of different subsets of peripheral T and NK cells in subjects were measured before and longitudinally after LDT treatment. Following treatment with LDT, the serum HBV DNA loads and the percentages of HBsAg- or HBeAg-seropositive cases were gradually reduced, accompanied by decreased levels of serum ALT and AST. In comparison with the HC, fewer CD3(-) CD56(+) and CD244(+) NK cells and CD3(+) CD8(+) T cells, lower frequencies of cytokine(+) CD4(+) T cells, and more CD3(+) CD4(+), CD4(+) CD25(+) Foxp3(+), CD4(+) CD25(+) CD127(low), and CD8(+) PD-1(+) T cells were detected in CHB patients. Treatment with LDT increased the numbers of NK and CD8(+) cells and the frequencies of cytokine(+) CD4(+) T cells but reduced the numbers of CD4(+) CD25(+) Foxp3(+), CD4(+) CD25(+) CD127(low), and CD8(+) PD-1(+) T cells in CHB patients. The frequencies of cytokine(+) CD4(+) T cells were negatively associated with the levels of serum HBV DNA, ALT, and AST. Thus, treatment with LDT inhibits HBV replication, modulates T and NK cell immunity, and improves liver function in Chinese patients with CHB.

Use of ALLGIO Probe Assays for Detection of HBV Resistance to Adefovir in Patients with Chronic Hepatitis B, Kerman, Iran

Hepatitis B virus (HBV) infection is contagious with transmissiobn vertically or horizontally by blood products and body secretions. Over 50% of Iranian carriers contracted the infection prenatally, making this the most likely route of transmission of HBV in Iran. To evaluate the resistance to adefovir (ADV) therapy in patients with chronic hepatitis B infection, a study was conducted on 70 patients (63 males and 7 females), who had received in first line lamivudine and second line adefovir. All were tested for the presence of hepatitis B surface antigen (HBsAg), hepatitis B envelope antigen (HBeAg), serum alanine amino transferase (ALT) level and HBV DNA load before and after treatment with ADV. In all samples, resistance to lamivudine and ADV was tested with real time PCR. Among seventy patients with chronic hepatitis B infection, 18 (25.7%) were resistant to LAM and 8 (11.4%) were resistant to ADV. Only one patient was negative for the presence of HBS-Ag (5.6%) and two were negative for HBe-Ag (11.1%). In this study we used a new method (ALLGIO probe assay) that has high sensitivity in detection of adefovir resistance mutants, which we recommend to other researchers. Mutant strains of the YMDD motif of HBV polymerase can be found in some patients under treatment with lamivudine and ADV. ADV has been demonstrated to be efficient in patients with lamivudine resistant HBV.

Hepatitis B Virus Infection Does Not Significantly Influence Plasmodium Parasite Density in Asymptomatic Infections in Ghanaian Transfusion Recipients

BackgroundAreas endemic for malaria and Hepatitis B virus (HBV) infection largely overlap geographically. A recent study has suggested the existence of an interaction between the two pathogens in symptomatic co-infected individuals on the South-American continent. We examined this issue in a hyperendemic area for both pathogens in sub-Saharan Africa.Methodology and FindingsPre-transfusion samples from a retrospective cohort of 154 blood transfusion recipients were screened for both serological and molecular markers of HBV and Plasmodium genomes using species-specific nested PCR and quantitative real-time PCR. Thirty-seven individuals met exclusion criteria and were subsequently eliminated from further analysis. Of 117 participants, 90% of recipients exhibited evidence of exposure to HBV, 42% with HBsAg and/or HBV DNA and 48% anti-HBc reactive without detectable HBV DNA. Plasmodium genome prevalence by NAT was 50%. Parasitemic individuals were significantly younger than non-parasitemic individuals (P = 0.04). Parasitemia level was not significantly lower in individuals with HBV DNA positive infections compared to those with HBV DNA negative exposures. HBV DNA load was not significantly different in parasitemic and non-parasitemic individuals.ConclusionThe data presented suggests that, in sub-Saharan Africa, asymptomatic co-infections with these two ubiquitous pathogens do not appear to significantly affect each other and evolve independently.

Pegylated interferon α enhances recovery of memory T cells in e antigen positive chronic hepatitis B patients

BackgroundInterferons (IFNs) are a group of cytokines commonly used in the clinical treatment of chronic hepatitis B (CHB) patients. Their therapeutic effects are highly correlated with recovery of host antiviral immunity. Clearance of hepatitis B virus (HBV) is mediated partially by activated functional memory T cells. The aims of the present study were to investigate memory T cell status in patients with different outcomes following pegylated interferon-α (IFN-α) therapy and to identify new biomarkers for predicting antiviral immune responses.MethodsPeripheral blood cells were isolated from 23 CHB patients who were treated with pegylated IFN-α at week 0 (baseline) and week 24. Co-expression of programmed death-1 (PD-1) and CD244 in CD45RO positive T cells, as well as a subset of CD127 and CXCR4 positive memory T cells were assessed. In addition, perforin, granzyme B, and interferon-γ (IFN-γ) expressions were also analyzed by flow cytometric analysis after intracytoplasmic cytokine staining (ICCS). Peripheral blood mononuclear cells (PBMC) isolated at week 24 were re-challenged with exogenous HBV core antigen, and the percentage of IFN-γ expression, serum HBV DNA loads, and ALT (alanine aminotransferase) levels were evaluated.ResultsAt week 24, PD-1 and CD244 expression in CD8 memory T cells were down-regulated (P < 0.05, P < 0.05, respectively), along with decreased HBV DNA loads (P < 0.05), while the expressions of partial effector molecules in CD8 and CD4 memory T cells was up-regulated (P < 0.05,P < 0.05, respectively), especially in the responders. CD127 and CXCR4 were highly expressed in CD8 memory T cells after pegylated IFN-α treatment (P < 0.05), which was inversely correlated with HBV DNA loads (r = −0.47, P = 0.001). The responders had a higher IFN-γ expression in memory T cells than the non-responders did after HBV antigen re-stimulation in vitro.ConclusionPegylated IFN-α treatment enhanced recovery of memory T cells in CHB patients by down-regulating inhibitory receptors and up-regulating effector molecules. The expressions of CXCR4 and CD127 in CD8 memory T cell may be used as biomarkers for predicting the outcome of treatment.

Hepatitis B Reactivation in a Renal Transplant Patient Due to a Surface Antigen Mutant Strain: A Case Report

Reactivation of hepatitis B virus (HBV) is a complication of immunosuppressive treatment in patients with a history of HBV exposure. Herein we have reported a case of reactivation after renal transplantation in a 52-year-old male chronic HBV carrier who was treated with hepatitis B immunoglobulin (HBIg) prophylaxis immediately after transplantation in addition to cyclosporine, mycophenolate mofetil and prednisolone for maintenance immunosuppression. After application of rituximab, the patient developed clinical hepatitis with a high load of HBV DNA. Sequence analysis of the surface (S) antigen corresponding to the amino acid residues 101-186 (including the a-determinant region) revealed a genotype D mutant strain, subtype ayw3 with a single amino acid substitution D144E within the S gene. This case suggested that immunosuppressive treatment enhanced with rituximab promoted the emergence of an HBV mutant within the determinant region of the S antigen, which escaped HBIg immunoprophylaxis causing HBV reactivation in a kidney transplant recipient.

Reduced Expression of Human Chemokine Genes RANTES and IP-10 in Hepatitis B Virus Mediated Cirrhosis of Liver

Regulated upon Activation Normal T cell Expressed and Secreted (RANTES) and interferon gamma inducible protein 10 (IP-10), both chemokines are chemotactic for immunocompetent cells and play an important role in cell mediated antiviral defense. The objective of this work was to assess the expression pattern of RANTES and IP-10 genes in peripheral blood mononuclear cells (PBMCs) of hepatitis B virus (HBV) infected patients having various disease severity. The study was performed on 79 HBV infected patients grouped into acute, inactive carriers (IC), chronic (CHB), cirrhosis and hepatocellular carcinoma (HCC) plus 41 healthy voluntary blood donors as controls. Quantification of HBV surface antigen (HBsAg) was done by a sandwich enzyme linked immunosorbent assay (ELISA). Conventional and real time polymerase chain reaction (PCR) were used for genotyping and determination of HBV DNA load respectively. RANTES and IP-10 mRNA expressions were evaluated by reverse transcription PCR (RT-PCR) and densitometry. Results obtained show that RANTES expression reduced significantly (p&lt;0.0001) in cirrhosis group in comparison to controls and remain unaltered in other disease categories. Reduction in IP-10 expression was significant (p=0.006) in patients of all disease categories than controls which was most evident in cirrhosis group (p&lt;0.0001). No association was found between the expression level of chemokines with HBV genotypes, HBsAg and HBV DNA levels in sera.It could be concluded that reduced expression of both the chemokines might be associated with lesser infiltration of immunocompetent cells to liver to avert further damage in cirrhosis.Serum level of both RANTES and IP-10 can be considered as prognostic marker of liver cirrhosis by validation studies.

Assessment of the factors associated with insulin resistance in patients with chronic hepatitis B infection

To investigate the factors associated with insulin resistance (IR) in patients with chronic hepatitis B virus (HBV) infection. Sixty-eight patients with mild chronic hepatitis B (MCHB) caused by HBV were recruited for study. Sixty-seven healthy individuals with no hepatitis virus infections and normal liver function were enrolled as controls. Demographic, anthropometric, clinical, and blood biochemical parameters were compared between the two groups. IR was determined by the homeostasis model assessment (HOMA-IR). The MCHB group was further divided into patients with IR (HOMA-IR: > 2.7) and patients without IR (HOMA-IR: less than 2.7). Demographic, anthropometric, clinical, and blood biochemical parameters were compared between the two sub-groups. Finally, the potential factors associated with IR were evaluated. Compared to the healthy controls, the MCHB patients had significantly higher serum insulin (Z = -5.451, P less than 0.01), alanine aminotransferase (ALT) (Z = -8.211, P less than 0.01) and HOMA-IR (Z = -5.631, P less than 0.01). IR was detected in 44.12% (30/68) of the MCHB patients. The levels of ALT and body mass index (BMI) were significantly different between the MCHB patients with IR and without IR (t = -2.358, and t = -3.566, P less than 0.05). There was a significant correlation between BMI, ALT, and HOMA-IR in the MCHB patients (r = 0.374, r = 0.282, P less than 0.05), but not with the HBV DNA loads (r = 0.015, P = 0.904). Binary logistic regression analysis indicated that BMI [Exp(B): 1.859, P less than 0.01] and ALT [Exp(B): 1.022, P less than 0.05] were independent risk factors of IR in MCHB. There is a high prevalence of insulin resistance in patients with mild hepatitis caused by chronic HBV infection. In these patients, IR is correlated with abnormal liver function and BMI, and not HBV load.