Stepwise Depletion of CD27+IgD+ B Cells Correlates With Fibrosis Progression to Cirrhosis in HBV-Infected Patients.

Background Anti-viral therapy is not routinely recommended for chronic hepatitis B virus (HBV) infection, in patients who have persistently elevated serum HBV DNA level (>2000 IU/mL), normal alanine aminotransferase (ALAT) and without significant liver fibrosis, defining the indeterminate phase. The objective of the study is to identify the factors associated with the progression of liver fibrosis in chronic HBV infected patients in the indeterminate phase. Methods This is cross-sectional study, conducted in Infectious Disease and Hepato-gastroenterology departments of Farhat Hached university hospital, between January 2008 and January 2022. We have included the Ag HBs (+) patients initially not treated, presenting at the time of the initial evaluation: a viral load> 2,000 IU/L for at least six months, normal ALAT (<40UI) and a fibrosis score F0 and/or F1 (in liverbiopsy or FibroScan). Univariate and logisticreg ression analysis were performed to identify the factors associated with liver fibrosis progression. Results In total, 97 patients were included, with a median age of 32.9± 9.1 years, and a female predominance (sex ratio M/F=0.64). Progression of fibrosis was observed in 16 patients (16.5%) with a mean delay of 70.9±41.1 months. In the univariate analysis, factors associated with progression of fibrosis were the presence of comorbidities (p=0.001), the high initial viral load (p=0.004), the appearance of cytolysis (p=0.001) and the increase in viral load (p=0.002), during follow-up. The AUROC of the initial viral load was 0.664 (95%CI: 0.500-0.820). An intial viral load at 8090 UI/l was associated with the progression of fibrosis with a sensibility of 70.3% and specificity of 63%. Conclusion Factors associated with progression of fibrosis in the indeterminate phase of chronic HBV infection were the presence of comorbidities, and changes of ALAT during follow-up. This leads us to consider extending the therapeutic indications to this group of patients.

Melanoma differentiation-associated protein 5 (MDA5) and retinoic acid-inducible gene 1 (RIG-1) as the pattern recognition receptors play important roles in viral mRNA recognition. Chronic HBV-infected (CHB) patients are unable to properly respond to hepatitis B virus (HBV). Therefore, the aim of the present study was to evaluate the mRNA levels of MDA5 and RIG-1 in the peripheral blood immune cells of CHB patients in comparison to healthy controls. In this cross-sectional study, the mRNA levels of MDA5 and RIG-1 were examined in 60 CHB patients and 60 healthy controls using the real-time polymerase chain reaction (PCR) technique. Our results showed that mRNA levels of MDA5 and RIG-1 were significantly decreased and increased, respectively, in CHB patients when compared to healthy controls. Our results also revealed that mRNA levels of MDA5 and RIG-1 were not altered among CHB patients with various states of e-antigen of hepatitis B and HBV-DNA viral loads. According to the results presented here, it may be concluded that downregulation of MDA5 may be a responsible mechanism from several reasons, which leads to HBV persistence in CHB patients.

Immune-mediated inflammatory injury is an important feature of the disease aggravation of hepatitis B virus-related acute-on-chronic liver failure (ACLF). Toll-like receptors (TLRs) have been shown previously to play a pivotal role in the activation of innate immunity. The purpose of this study was to characterize the TLR4 expression in peripheral blood mononuclear cells (PBMCs) of ACLF patients and its possible role in the disease aggravation. Twelve healthy subjects, 15 chronic HBV-infected (CHB) patients and 15 ACLF patients were enrolled in this study. The TLR4 expression in PBMCs and T cells of all subjects was examined by real-time PCR and flow cytometry. The correlation of TLR4 expression on T cells with the markers of disease aggravation was evaluated in ACLF patients. The ability of TLR4 ligands stimulation to induce inflammatory cytokine production in ACLF patients was analyzed by flow cytometry. The results showed that TLR4 mRNA level was upregulated in PBMCs of ACLF patients compared to that in the healthy subjects and the CHB patients. Specifically, the expression of TLR4 on CD4(+) and CD8(+) T cells of PBMCs was significantly increased in ACLF patients. The TLR4 levels on CD4(+) and CD8(+) T cells were positively correlated with serum total bilirubin (TBIL), direct bilirubin (DBIL), international normalized ratio (INR) levels and white blood cells (WBCs), and negatively correlated with serum albumin (ALB) levels in the HBV-infected patients, indicating TLR4 pathway may play a role in the disease aggravation of ACLF. In vitro TLR4 ligand stimulation on PBMCs of ACLF patients induced a strong TNF-α production by CD4(+) T cells, which was also positively correlated with the serum markers for liver injury severity. It was concluded that TLR4 expression is upregulated on T cells in PBMCs, which is associated with the aggravation of ACLF.

Hepatitis B is one of the most common infectious diseases in China, and it is necessary to study biochemical indicators of HBV infected patients. Biochemical and basic features of 1765 HBV infected patients and 840 general controls were collected and analyzed from Southwest China. A total of 23.5% of the patients were children or young persons (less than 40 years old). The co-infected rate was only 0.3% for those who were co-infected with Hepatitis C virus (HCV) or Treponema pallidum (TPa). Most of patients (1081 individuals) were under convalescent condition, and 4.2% and 34.6% HBV patients were at acute and chronic phase, respectively. The indicators of liver function were significantly different between HBV patients and normal controls. In order to further study the variation of biochemical features in HBV patients in different phase, they were divided into three groups (sample #1: patients in acute infected phase; sample #2: patients in chronic infected phase; and sample #3: patients in convalescent condition). Excluding total protein and globin, all other indicators of liver function were statically different among the three groups. Total protein and albumin gradually decreased from convalescent patients, chronic HBV-infected patients, to acute HBV-infected patients. Biochemical features could be used to evaluate the progress and therapeutic effects of HBV-infection. Our analysis firstly reported basic and biochemical information of HBV patients in Southwest China.

ObjectiveTo investigate the impact of SARS-CoV-2 infection on liver function and prognosis in patients with HBV infection.MethodsA total of 154 HBV-positive patients (HBV ( +) group) and 154 HBV-negative patients (HBV (-) group) diagnosed with COVID-19 at Taizhou Hospital between December 10, 2022, and January 31, 2023, were included in this study. Clinical characteristics, treatment, and laboratory findings were collected from patients at three time points: before (T1), during (T2), and at the time of discharge (T3) from SARS-CoV-2 infection.ResultsCompared to the HBV (-) group, the HBV ( +) group had a longer hospital stay (15 (9–22) days vs. 9 (5–16) days). Longitudinal comparisons of laboratory indicators from T1 to T3 showed a continuous decline in TP and ALB levels and a continuous increase in PT and TT levels in the HBV ( +) group. BUN levels increased during T2 and decreased thereafter. These differences were considered statistically significant (P < 0.05). Notably, the HBV ( +) group had a higher proportion of indicators elevated > 3 ULN from T1 to T2, including ALT (1.95%/5.19%), AST (3.25%/12.99%), ALP (1.95%/3.25%), GGT (4.55%/9.09%), TBIL (6.49%/9.09%), and DBIL (18.18%/22.73%). In the HBV (-) group, the elevations were mainly concentrated within 1–2 ULN, including AST (12.99%/22.08%), DBIL (10.39%/21.43%), BUN (12.99%/22.08%), CREA (20.13%/29.22%), and PLT (7.79%/14.94%). Furthermore, the incidence of liver injury from T1 to T3 was higher in the HBV ( +) group compared to the HBV (-) group (15.7% (20/127) vs. 7.2% (11/152), P < 0.05). Multivariate analysis showed that liver cirrhosis (HR = 4.847, 95% CI: 1.224–19.20, P = 0.025) and liver cancer (HR = 8.333, 95% CI: 2.156–32.209, P = 0.002) were independent risk factors for liver injury in the presence of SARS-CoV-2 infection.ConclusionSARS-CoV-2 infection has a higher proportion of liver injury in HBV-infected patients, affecting hepatic protein synthesis function. Those with cirrhosis and hepatocellular carcinoma are at higher risk of severe liver injury.

Some single nucleotide polymorphisms (SNPs) of the gene encoding interleukin 28B (IL28B) may increase susceptibility to infection and chronicity in humans with hepatitis B and C viruses. In our study, we aimed to investigate the prevalence of rs12979860, rs8099917 and rs12980275 SNPs in IL28B in patients with hepatitis B (HBV) or hepatitis C Virus (HCV) infection and to determine the relationship of these polymorphisms with plasma IL28B levels. For this purpose, 64 HBV-infected and 66 HCV-infected patients and 70 healthy individuals were included in the study. The SNPs were investigated by real time PCR (Polimerase Chain Reaction, Rt-PCR) using TaqMan SNP Genotyping Assay. The plasma levels of IL28B were detected by 'Enzyme Linked Immunosorbent Assay (ELISA)'. The frequencies of the rs12980275AG genotype and G allele (p= 0.003 and p= 0.04, respectively), and the rs12979860CT genotype and T allele (p= 0.01 and p= 0.04, respectively) were lower in HBV-infected patients. In HCV-infected patients, the rs8099917TG genotype and G allele frequencies (p= 0.04) were higher and the TGG haplotype showed a statistically significant difference (p= 0.04). The mean of IL28B plasma levels were higher in the control group than the HBV or HCV-infected patient groups (p= 0.001 and p= 0.01, respectively). However, HBV-infected patients with the rs12980275AG genotype showed a significant difference in plasma IL28B levels compared to the other genotypes (p= 0.0001) and these patients had lower viral loads (<105 IU/ml). According to the results of the study, it can be stated that rs12979860CT and rs12980275AG genotypes may play a role in preventing the chronicity of HBV infection, while rs8099917TG genotype may contribute to the transformation of HCV infection into chronic infection. In this study, it was observed that the presence of the G allele for the rs8099917 polymorphism could be evaluated as a risk allele for chronic HCV infection and that the TGG haplotype could have a strong predictive effect on increasing susceptibility to chronic HCV infection. It is recommended to evaluate the genotypic distribution of IL28B before treatment because of its prognostic significance in HBV or HCV infected patients. In HBV infection, the rs12980275AG genotype which is thought to have a protective effect by limiting viral replication with increased plasma IL28B levels, can be used as a good prognostic factor. These polymorphisms could be used as biomarkers to predict the clinical consequences of the patients infected with HBV or HCV, to take precautions to prevent the chronicity of the infection and its complications, and to develop new molecular targeted therapies with further research.

Expression profiles and function of Toll-like receptors 2 and 4 in peripheral blood mononuclear cells of chronic hepatitis B patients

Background: Depression and anxiety can modulate immune-related molecule expressions. The chronic HBV-infected (CHB) patients suffer from inappropriate immune responses. Additionally, psychological disorders are prevalent among the patients. Thus, depression and anxiety may alter immune-related molecule expression. This study aimed to examine IPS-1 and RIP1 mRNA levels in CHB patients suffering from various degrees of anxiety and depression. Methods: Sixty patients with CHB participated in this research and completed standard questionnaires to evaluate depression and anxiety. The expression levels of IPS-1 and RIP1 were examined using real-time PCR techniques. Results: The result revealed that although the expression of IPS-1 and RIP1 did not change in the CHB patients with various ranges of depression and anxiety, IPS-1 was significantly decreased in the male CHB patients who suffered from mild, moderate, and severe depression when compared to the patients with no depression. Conclusion: So, it was hypothesized that depression may be associated with alteration in the expression of IPS-1 in a sex-dependent manner. In other words, it appears that the male CHB patients are at risk of depression-related alteration in immune-related gene expression.

Hepatitis B virus (HBV) infection is considered as one of the most serious public health problems worldwide including Egypt. Soluble fibrinogen-like protein 2 (sFGL2) is a well-known immunomodulator that is produced by the T cells and has a strong inhibitory effect on the proliferation of T cells and maturation of dendritic cells (DC). In the current study, serum levels of sFGL2 were assessed utilizing enzyme-linked immunosorbent assay (ELISA) technique among 20 acute HBV-infected patients, 55 chronic HBV-infected patients and 15 healthy individuals. In addition, serum levels of soluble FAS ligand (sFASL), soluble FAS receptor (sFAS) as well as interferon-γ (IFN-γ) were assessed and correlated to the levels of sFGL2. According to our results, serum levels of sFGL2 were significantly higher in the acute HBV-infected patients than in the chronic HBV-infected patients and healthy individuals. On the other hand, the serum levels of sFASL, sFAS and IFN-γ were significantly higher in the chronic than in acute HBV-infected patients. Also, serum sFGL2 levels were negatively correlated with the serum levels of sFASL, sFAS, IFN-γ and albumin as well as hemoglobin concentration. Furthermore, serum sFGL2 levels were positively correlated with the activities of ALT and AST and total bilirubin levels in serum. Thus, the current work highlights the possibility of utilizing serum sFGL2 level as a novel biomarker for the differentiation between acute and chronic Egyptian HBV-infected patients.

To estimate the prevalence of diagnosed and undiagnosed coinfections among HIV, HBV, and HCV infected patients. Retrospective analysis of laboratory records for HIV, HBV, and HCV patients presenting at the HIV outpatient clinic. Serological data including hepatitis B surface antigen (HBsAg), hepatitis B e-antigen (HBeAg), hepatitis B e-antibody (anti-HBe), antibodies to HIV and HCV, anti-toxoplasmosis IgG and IgM antibodies, and anti-syphilis antibodies (VDRL) were collected. We obtained data for 628 (218 HCV, 268 HBV, and 142 HIV) patients. Male-to-female ratios were 1:1 for HCV, 3:4 for HBV, and 5:3 for HIV. Age means (SD) were 54.24 (16.40), 44.53 (18.83), and 40.39 (15.92) years for HCV, HBV, and HIV, respectively. In HIV group, the prevalence of HBV and HCV coinfections was 8.5% and 2.8%, respectively. In HBV group, the prevalence of HCV and HIV coinfections was 1.1% and 1.5%, respectively. In HCV group, HIV or HBV coinfections occurred at the same frequency (1.4%). An absence of screening for coinfections was detected in 7.0-48.5% patients as per the group and the infectious agent; which represents an estimated proportion of 20 out of 1,000 patients with an undiagnosed coinfection. Despite a relatively low prevalence of coinfections, a significant proportion of cases remain undiagnosed because of a lack of systematic screening. J. Med. Virol. 88:1545-1551, 2016. © 2016 Wiley Periodicals, Inc.

Gut microbiota dysbiosis has been observed in HBV-related cirrhosis, but its role in early-stage disease and its correlation with liver pathology remain unclear. Moreover, whether dysbiosis is a cause or consequence of liver cirrhosis is still debated. We recruited 20 treatment-naïve patients with chronic HBV infection, assessing liver injury via biopsy. Fecal metagenomic sequencing was used to analyze the correlation between gut microbiota and liver histology. To explore the causality, fecal samples from an HBV-related cirrhosis patient were transplanted into mice with CCl₄-induced liver fibrosis. Patients with significant histological damage exhibited reduced alpha diversity and greater microbial homogeneity. Species such as Eubacterium_sp_CAG_180, Gemmiger_formicilis, and Oscillibacter_sp_ER4 had decreased abundance, while Parabacteroides_distasonis, Bacteroides_dorei, and Bacteroides_finegoldii were enriched. Mice receiving fecal transplants from the cirrhotic patient showed aggravated liver fibrosis, with increased collagen deposition; elevated ALT, AST, and ALP levels; and heightened hepatic inflammatory gene expression. Additionally, abnormal bile acid profiles with elevated unconjugated bile acids (e.g., GCA and CA) were observed. Gut microbiota dysbiosis is closely associated with liver histological damage in chronic HBV infection and may drive fibrosis progression via microbial-bile acid interactions. These findings suggest potential for gut microbiota-based assessment and treatment strategies in chronic hepatitis B.IMPORTANCEThis study elucidates a significant association between gut microbiota dysbiosis and liver histological damage in patients with chronic hepatitis B (HBV), potentially exacerbating fibrosis progression through bile acid interactions. By analyzing patient gut microbiota and conducting fecal transplant experiments in mice, researchers have identified that gut microbiota dysbiosis contributes to hepatic fibrosis during chronic HBV infection. These findings underscore the importance of the gut-liver axis in HBV disease progression, indicating that monitoring or modulating gut bacteria may facilitate early diagnosis or therapeutic interventions. This research bridges the gap in understanding whether microbial alterations drive disease progression or result from it, providing a foundation for developing therapies targeting the microbiome to mitigate liver damage in chronic HBV infections.

Background: Toll- like receptors (TLRs) play an important role in the recognition of DAMPs andPAMPs and induction of inflammation. Previous studies demonstrated that depression and anxietycan influence the expression levels of immune related molecules. Our previous study revealed thatmRNA levels of IRAKIRAK4, TRAF3 and IRF7 were significantly decreased in chronic HBV infected(CHB) patients when compared to healthy controls. Therefore, the aim of this study was toevaluate the effects of depression and anxiety on the expression levels of these molecules in CHBpatients.Methods: Sixty CHB patients participated in this study and filled out the standard questionnaires;and the expression of IRAK4, TRAF3 and IRF7 were examined using Real-Time PCR techniques.Results: The results of this study demonstrated that expression of IRAK4, TRAF3 and IRF7 did notdiffer between patients with various stages of depression and anxiety (all p>0.05).Conclusion: According to the results, it seems that declined expression of IRAK4, TRAF3 andIRF7 in CHB patients were not related to depression and anxiety, and other factors including geneticand immunoregulatory effects of HBV may be responsible for the declined expression of these molecules.

Decreased expression of toll like receptor signaling molecules in chronic HBV infected patients

To evaluate the correlation between the hepatic expression pattern of hsa-miR-125a-5p and HBV-DNA and the progression of fibrosis in patients with overt or occult HBV infection. We enrolled all the HBsAg-positive treatment naive patients (overt HBV group) and all the HBsAg-negative patients with hepatocellular carcinoma and with a positive HBV-DNA in their hepatic tissue (occult HBV group), who underwent a diagnostic liver biopsy between April 2007 and April 2015. Tissue concentrations of HBV-DNA and hsa-miR-125a-5p were then analyzed by real-time quantitative PCR. Necroinflammatory activity and fibrosis were evaluated according to the Ishak score. During the study period, we enrolled 64 patients with overt and 10 patients with occult HBV infection. In the overt HBV group, 35 of 64 (54.7%) showed a mild fibrosis (staging 0-2), 17 (26.6%) a moderate fibrosis (staging 3-4), while the remaining 12 (18.7%) had a cirrhosis. All patients in the occult HBV group were cirrhotic. Patients with more advanced fibrosis stage showed a higher mean age when compared with those with mild (p < 0.00001) or moderate fibrosis (p < 0.00001) and were more frequently male than patients with staging 0-2 (p = 0.04). Similarly, patients with occult B infection were older than HBsAg-positive patients. Liver concentrations of miR-125a-5p were significantly higher in patients with cirrhosis (9.75 ± 4.42 AU) when compared with patients with mild (1.39 ± 0.94, p = 0.0002) or moderate fibrosis (2.43 ± 2.18, p = 0.0006) and were moderately higher in occult than in overt HBV infection (p = 0.09). Moreover, we found an inverse correlation, although not statistically significant, between the tissue HBV-DNA levels and the staging of fibrosis. This study suggests a correlation between the tissue expression of hsa-miR-125a-5p and the progression of liver damage in a group of patients with occult or overt HBV infection. If confirmed, these data suggest the hsa-miR-125a-5p may be a novel biomarker of hepatic damage.

Fibrosis is a hallmark histologic event of viral hepatitis and is characterized by the excessive accumulation and reorganization of the extracellular matrix (ECM). The gold standard for assessment of fibrosis is liver biopsy. As this procedure has various limitations, including risk of patient injury and sampling error. Serum Hyaluronic acid as non invasive marker for liver fibrosis is desirable. The present study aims to determine the serum hyaluronic acid (HA) levels as biochemical marker of hepatic fibrosis and cirrhosis and correlate it with the degree of hepatic fibrosis. Serum HA level in chronic hepatitis patients (n=60) are divided into two groups, group1: included 30 patients positive for anti-HCV (antibodies), group 2 included 30 patients positive for HBsAg, and controls (n=10) were assessed by ELISA and liver histopathological parameters were evaluated by the modified Knodell score and microscopic examination of liver biopsies from patients. Individuals in healthy control group have normal levels of HA (mean 14.3, SD: 5.5) while the levels of HA were elevated in patients of HCV alone (mean 103.6±28.0) and in patient of HCV (mean 104.5± 37.5).Also levels of HA were poorly elevated in HBV alone (mean 62.2± 15.5) and in HBV (mean 45.8± 12.4). showed that serum HA levels are well correlated with HAI in patients of HBV & HCV groups where, there was significant increase in HA levels by increase of HAI by liver biopsy P < 0.001.HA levels and stages of fibrosis were well correlated in patients of HBV and HCV group. Where, this is a significant increase in HA levels when Considering F0 to F6 scores by liver biopsy (P < 0.001). Serum HA is a useful non-invasive marker of liver fibrosis. There is a strong positive correlation between serum HA levels and degree of liver fibrosis. The concentration of serum HA rises according to progression of liver fibrosis.