Soluble programmed death ligand 1 (sPD-L1) reflects immune–inflammatory activity and may serve as a circulating biomarker of hepatocarcinogenesis. Whether baseline sPD-L1 predicts de novo hepatocellular carcinoma (HCC) in patients without existing HCC remains unclear. To determine whether serum sPD-L1 predicts future HCC across distinct virological backgrounds of chronic hepatitis C. Two temporally and clinically distinct cohorts from Nagasaki Medical Center were analyzed. The SVR cohort included 430 patients who achieved sustained virological response after direct-acting antiviral therapy (2013–2017). A historical pre-DAA cohort (n = 692), consisting of viremic patients who underwent liver biopsy between 1992 and 2003, served as an independent temporal validation cohort. Baseline sPD-L1 was measured in all patients. Cox proportional hazards models, stratified analyses, landmark analysis, and time-dependent models were applied to assess the association between sPD-L1 and HCC. High baseline sPD-L1 independently predicted increased HCC risk in both cohorts (SVR: HR 2.61, 95

AIM:To evaluate the effectiveness of prevention of mother-to-child transmission (PMTCT) over 25 years, we conducted a nationwide epidemiological survey through a comprehensive assessment of hepatitis B virus (HBV) markers. METHODS:Two cohorts were recruited before universal vaccination: children aged 9-10 years undergoing school health checkups (Group 1) and hospital patients 0-15 years of age from 11 hospitals across Japan to assess age and regional differences (Group 2). Hepatitis B surface antigen (HBsAg) and core antibody (anti-HBc) were measured in both cohorts. HBV-DNA was tested to evaluate occult HBV infection and viral genotypes in Group 2. Anti-HBs antibody was measured to estimate HBV vaccine coverage. RESULTS:In Group 1 and Group 2, 8125 and 8453 samples were collected, respectively. HBsAg prevalence was 0% in Group 1 and 0.047% in Group 2 using a conventional assay (Lumipulse II HBsAg). Most HBsAg-positive samples were from children < 4 years. HBsAg-negative and anti-HBc-positive samples were detected in 0.42% of Group 1 and 0.95% of Group 2. Among these samples in Group 2, 10.3% were HBV-DNA-positive, indicating occult HBV infection. Genotype C predominated; however, four genotype D cases were identified in Hokkaido. Anti-HBs positivity among HBV-uninfected samples at ages 1, 2, 5, 10, and 15 years was 53.4%, 27.5%, 7.5%, 5.6%, and 1.9%, respectively. CONCLUSIONS:Following the implementation of PMTCT measures in Japan, the prevalence of HBsAg-positive children was extremely low. However, the prevalence of occult infection and anti-HBc positivity indicated that some children remained exposed to HBV. To further eradicate HBV infection, universal vaccination was introduced nationwide in 2016.

Hepatitis B virus (HBV) reactivation is a well-recognized complication in patients with resolved HBV infection undergoing allogeneic hematopoietic cell transplantation (allo-HCT). Although HBV DNA-guided preemptive nucleos(t)ide analog therapy prevents HBV-related hepatitis, strategies to prevent HBV reactivation itself remain limited. Hepatitis B vaccination after allo-HCT has been suggested as a potential preventive strategy, but prospective randomized evidence is lacking. To evaluate whether hepatitis B (HepB) vaccination reduces HBV reactivation after allo-HCT and to examine the association between vaccination-induced anti-hepatitis B surface antibody (anti-HBs) response and protection against HBV reactivation. In this multicenter randomized controlled trial (UMIN000034113), adults who were hepatitis B surface antigen-negative and anti-hepatitis B core antigen-positive and underwent first allo-HCT were prospectively enrolled. At 140 days post-HCT, patients without HBV reactivation were randomized to receive HepB vaccination or no vaccination. HBV DNA levels and anti-HBs titers were centrally monitored for 2 years after transplantation. The primary endpoint was HBV reactivation, defined as HBV DNA ≥ 1.3 log IU/mL. Vaccine response was defined as anti-HBs response, requiring both an increase in anti-HBs from baseline and a post-vaccination anti-HBs ≥ 10 mIU/mL. Among 109 enrolled patients, 64 were randomized. With a median HBV DNA follow-up of 19.4 months, HBV reactivation occurred in 6 of 33 patients in the vaccination group, including 2 who experienced reactivation before receiving the first vaccine dose, and 9 of 31 patients in the control group. Probabilities of HBV reactivation were 19.8% and 37.9% in the HepB vaccination and control groups, respectively, at 18 months after randomization (P = .205). Baseline anti-HBs <10 mIU/mL was a significant risk factor for HBV reactivation. No HBV-related hepatitis was observed. Vaccine responses were achieved in 9 of 29 patients after the initial series and in 5 of 10 after the additional series. Importantly, HBV reactivation did not occur after patients achieved anti-HBs response, although several reactivation events occurred before vaccination or before anti-HBs response was achieved. No severe vaccine-related adverse events were observed. Although HepB vaccination did not significantly reduce HBV reactivation, it was feasible and induced anti-HBs response in a subset of patients. Achieving vaccine-induced anti-HBs response may provide protective immunity against HBV reactivation after allo-HCT.

BACKGROUND & AIMS:Hepatitis B surface antigen (HBsAg) seroclearance is widely regarded as a functional remission of chronic hepatitis B virus (HBV) infection. However, HBV covalently closed circular DNA (cccDNA) may persist in hepatocytes, posing a risk of reactivation even years after seroclearance. We aimed to determine the incidence, clinical context, and virological features of HBV reactivation in a large, population-based cohort from a geographically isolated island region of Japan with a high HBV prevalence. METHODS:Between 1978 and 2008, 34,517 residents of the northern Goto Islands underwent community-based HBsAg screening, identifying 1045 individuals with chronic HBV infection (3.0% prevalence). Among them, 177 achieved spontaneous HBsAg seroclearance and were followed for a median of 6.3 years (1500 person-years). Age- and sex-matched controls were selected from HBsAg-negative residents (3043 person-years). Reactivation-related events were defined as reappearance of HBsAg after confirmed seroclearance. RESULTS:HBV reactivation-related events occurred in 4 of 177 patients (2.3%), corresponding to 27 per 10,000 person-years, whereas none occurred among 354 matched controls. All HBsAg reappearance cases had cirrhosis or hepatocellular carcinoma at baseline. One patient developed a hepatitis flare with high-level viremia and HBeAg reappearance. Viral sequencing demonstrated 99.1% similarity between baseline and reactivated strains, confirming reactivation rather than reinfection. CONCLUSIONS:Reactivation-related events after HBsAg seroclearance were rare in this population-based cohort. These findings suggest that HBsAg seroclearance may not necessarily indicate complete viral eradication and that careful long-term monitoring may be warranted, particularly in patients with advanced liver disease.

Hepatitis B virus (HBV) infects hepatocytes by using sodium taurocholate cotransporting polypeptide (NTCP) as a receptor. NTCP can render non-permissive hepatocyte cell lines such as HepG2 permissive to HBV infection. However, only a few reports investigate the NTCP function in HBV infection beyond virus binding. Here, we performed immunopurification followed by liquid chromatography-tandem mass spectrometry to identify ZO-2 (aka TJP2) as a novel NTCP-binding protein. Knockdown or knockout of ZO-2 in NTCP-transduced HepG2 cells decreased the amount of NTCP at the cell surface, leading to reductions in cellular attachment and infection by HBV. Incubation of cells with HBV surface molecules preS1 resulted in the dissociation of NTCP from ZO-2 and in the formation of NTCP-preS1-actin complexes that were internalized into the cell. Latrunculin A, an inhibitor of actin polymerization, suppressed the preS1 internalization into hepatocytes and HBV infection. In conclusion, ZO-2, together with actin, regulates the function of NTCP as a receptor of HBV, providing a new model for HBV-cell interactions and virus internalization. IMPORTANCEAlthough a number of candidates have been reported to bind to the hepatitis B virus (HBV) envelope, accumulating evidence indicates that NTCP is accepted as a functional receptor for HBV infection. Thus, NTCP is an attractive target for antiviral therapies. Here, we showed that ZO-2 interacts with NTCP. The silencing of ZO-2 decreased HBV infection, whereas ZO-1 and ZO-3 knockdown had no effect on HBV infection. Moreover, knockout of ZO-2 induced the downregulation of NTCP from the cell surface. This aberrant NTCP localization causes the reduction of the half-life of NTCP in ZO-2 knockout cells. PreS1 treatment or HBV infection disrupted the NTCP/ZO-2 complex through the dissociation of the actin-binding domain of ZO-2, leading to internalization of a newly formed preS1/NTCP/actin complex into the cell. The actin polymerization inhibitor latrunculin A suppressed HBV infection. These results suggest that ZO-2 regulates cell surface localization of NTCP.

Hepatitis B virus (HBV) infects human populations worldwide. HBV strains are classified into 10 genotypes, of which the HBV genotype D (HBV/D) infection is particularly prevalent in several countries. The HBV core promoter regulates viral replication and transcription, and the naturally occurring A1762T/G1764A double mutation (CP1) in the core promoter accelerates HBV replication. Previous clinical studies showed that a new core-promoter mutation, G1764T/C1766G (CP2), is frequently observed in genomes containing the G1757A substitution, which is unique to HBV/D; however, CP2 is not observed in genomes containing the 1762T/1764A double mutation. In this study, we found that the CP2 mutation dramatically increased viral replication and transcription efficiency in two cell lines; the degree of stimulation was comparable to that induced by CP1. Introduction of the 1757A substitution reduced the increase in viral replication induced by the CP1 mutation. By contrast, the addition of the 1757A substitution significantly increased the effect of the CP2 mutation. The transcriptional activity of CP1 was decreased by the 1757A substitution, due to a reduction in HNF1 binding affinity, suggesting that 1757G is an important component of the HNF1 binding consensus sequence. The HBV/D-specific CP2 mutation creates a binding site for the transcription factor HNF3, thereby increasing its transcriptional activity. HBX proteins containing substitutions reflecting the two types of core-promoter mutations did not affect the efficiency of viral replication. Therefore, we hypothesize that the introduction of the CP2 mutation represents a survival strategy for HBV/D, allowing it to escape the effect of the 1757A substitution.

BACKGROUND AND AIMS:Various viral mutations have been reported to influence disease pathogenesis in infectious diseases. However, these mutations may remain undetected due to patient population changes. Although many causes have been proposed, the exact reasons remain unclear. This study analyzed viral mutations linked to hepatocellular carcinoma (HCC) in patients with hepatitis B by comparing factors between condition-matched patient groups of different human leukocyte antigen (HLA) genotypes. METHODS:HLA genotypes of 2281 healthy individuals, 370 patients with hepatitis B virus (HBV)-derived HCC, and 408 patients with chronic hepatitis B were analyzed to identify HLA genotypes associated with HCC development. Patients with HCC were grouped by HLA-DPB1 genotype to identify HBV genomic variants linked to HCC. RESULTS:The HLA-DPB1*02:01, 04:01, and 04:02 alleles suppressed chronic hepatitis B (CHB) and HCC development (P = 4.53 × 10-9, 3.20 × 10-3, and 3.03 × 10-4, respectively). Conversely, HLA-DPB1*05:01 and 09:01 alleles were significantly associated with CHB and HCC development (P = 1.01 × 10-7, 6.51 × 10-8, respectively). Among HLA-DPB1*02:01 patients, the S166L amino acid (AA) mutation in the PreS/S region of the HBV genome was significantly identified. HLA-DPB1*05:01 homozygous patients significantly had K130M/V131I AA mutations in the X region of the HBV genome, whereas HLA-DPB1*05:01-09:01 heterozygous patients significantly had the H94Y AA mutation in the X region. Stratification by HLA genotype changed the mutation position and strengthened the odds ratios for developing HCC due to viral mutations. CONCLUSIONS:Our analysis revealed that viral mutations associated with HCC vary by HLA genotype. These findings suggest a host-pathogen genetic interaction. HLA genotype stratification may aid in advancing personalized genomic medicine, which has been challenging to apply to chronic diseases.

A subset of patients still develops hepatocellular carcinoma (HCC) even after eradication of the hepatitis-C virus (HCV) by anti-HCV treatment. We conducted a genome-wide association study (GWAS) to identify host genetic factors associated with HCC development following HCV eradication in Japan. In this GWAS (n = 517), the discovery cohort included 118 patients without HCC and 67 who developed HCC following HCV eradication with interferon-based therapy. A genome-wide scan for HCC-associated variants was conducted. An independent cohort of 274 patients without HCC and 58 patients with post-eradication HCC was used for replication. The effects of candidate gene variants were assessed clinically and through in vitro cellular assays. The GWAS identified significant variants associated with HCC development following HCV eradication, including rs4778350, located near the long non-coding RNA Prader-Willi non-protein coding RNA 4 (PWRN4) on chromosome 15. In the combined analysis, rs4778350 remained significantly associated with HCC, showing a high odds ratio of 5.86 (95

A cure for chronic hepatitis B requires eliminating or permanently silencing covalently closed circular DNA (cccDNA). A pivotal target of this approach is the hepatitis B virus (HBV) X protein (HBx), which is a key factor that promotes transcription from cccDNA. However, the HBx structure remains unsolved. Here, we present the cryoelectron microscopy structure of HBx in complex with DDB1, which is an essential complex for cccDNA transcription. In this structure, hydrophobic interactions within HBx were identified, and mutational analysis highlighted their importance in the HBV life cycle. Our biochemical analysis revealed that the HBx-DDB1 complex directly interacts simultaneously with NSE3, which is a component of the SMC5/6 complex, and Spindlin1. Additionally, HBx-DDB1 complex dynamics were explored via high-speed atomic force microscopy. These findings provide comprehensive insights into the structure and function of HBx in HBV replication.

BACKGROUND AND AIMS:HBV leads to severe liver diseases, such as cirrhosis and HCC. Identification of host factors that regulate HBV replication can provide new therapeutic targets. The discovery of sodium taurocholate cotransporting polypeptide (NTCP) as an HBV entry receptor has enabled the establishment of hepatic cell lines for analyzing HBV infection and propagation. Using this new system, studies aimed at identifying host factors that regulate HBV propagation have increased. APPROACH AND RESULTS:We established an HBV-based-reporter gene expression system that mimics HBV replication from transcription to virus egress. Using this approach, we screened 1827 Food and Drug Administration-approved compounds and identified glycogen synthase kinase 3 (GSK3)alpha/beta inhibitors, including AZD1080, CHIR-98014, CHIR-98021, BIO, and AZD2858, as anti-HBV compounds. These compounds suppressed HBeAg and HBsAg production in HBV-infected human primary hepatocytes. Proteome analysis revealed that GSK3alpha/beta phosphorylated forkhead box K1/2 (FOXK1/2)s. A double-knockout of FOXK1/2 in HBV-infected HepG2-NTCP cells reduced HBeAg and HBsAg production. The rescue of FOXK2 expression, but not FOXK1 expression, in FOXK1/2-double-knockout cells restored HBeAg and HBsAg production. Importantly, phosphorylation of FOXK2 at Ser 424 is required for GSK3alpha/beta-mediated HBeAg and HBsAg production. We observed the binding of FOXK2 to HBV DNA in HepG2-NTCP cells. CONCLUSIONS:Our recombinant HBV-based screening system enables the discovery of new targets. Using our approach, we identified GSK3 inhibitors as potential anti-HBV agents.

Abstract Background/purpose of the study: Mac-2-binding protein glycosylation isomer (M2BPGi), a biomarker for liver fibrosis, is influenced by various etiologies. Here, we aimed to investigate clinical factors that improve the accuracy of liver cirrhosis (LC) diagnosis based on quantitative M2BPGi (M2BPGi-Qt), regardless of etiology. Methods: In total, 1,373 patients with chronic liver disease (CLD) were recruited. Weassessed the correlation between fibrosis stage and M2BPGi-Qt levels among CLD etiologies. If there was no correlation between the fibrosis stage and M2BPGi level in a specific etiology of CLD, we evaluated the clinical factors influencing the M2BPGi-Qt level in that specific etiology. Subsequently, we created an algorithm to detect LC based on M2BPGi-Qt, considering an influencing factor other than fibrosis. Results: In virus hepatitis, non-alcoholic fatty liver disease, and primary biliary cholangitis, the M2BPGi-Qt levels increased liver fibrosis progression. In autoimmune hepatitis, no significant association was observed between the fibrosis stage and M2BPGi-Qt level. However, liver inflammation positively correlated with the M2BPGi-Qt levels. Considering liver inflammation, we established an algorithm, M2BPGi-Qt, to determine the alanine aminotransferase-to-platelet ratio (MAP-R) in LC. The area under the receiver operating characteristic curve (AUC) of the MAP-R index was 0.840. The AUC of MAP-R was higher than that of the M2BPGi-Qt for detecting LC. Conclusions: New quantitative measurement system for M2BPGi reveals liver inflammation complicates liver cirrhosis diagnosis. The algorithm based on the M2BPGi-Qt level demonstrates a high accuracy for LC diagnosis.

The relationship between liver fibrosis and inflammation and Mac-2-binding protein glycosylation isomer (M2BPGi) in patients with chronic liver disease (CLD) other than hepatitis C remains uncertain, owing to the limitations of qualitative methods. Here, we evaluated the influence of liver fibrosis and inflammation on quantitative M2BPGi (M2BPGi-Qt) in CLD, considering each etiology. We recruited 1373 patients with CLD. To evaluate the influence of liver fibrosis and inflammation on M2BPGi-Qt levels, we assessed M2BPGi-Qt levels at each fibrosis and activity stage within different etiologies of CLD based on pathological findings. Subsequently, we evaluated if the accuracy of fibrosis staging based on M2BPGi-Qt could be improved by considering the influence of liver inflammation. In patients with viral hepatitis, non-alcoholic fatty liver disease, and primary biliary cholangitis, the median M2BPGi-Qt levels increased liver fibrosis progression. Median M2BPGi-Qt levels were not associated with the degree of fibrosis in patients with autoimmune hepatitis (AIH). Median M2BPGi-Qt levels increased with the progression of liver activity in all etiologies. A significant difference was found at each stage in AIH. Considering the liver inflammation, we established an algorithm, M2BPGi-Qt, to determine the alanine aminotransferase-to-platelet ratio (MAP-R) in liver cirrhosis (LC). The area under the receiver operating characteristic curve (AUC) of MAP-R was higher than that of the M2BPGi-Qt for detecting LC (AUC MAP-R = 0.759 and M2BPGi-Qt = 0.700, p < 0.001). The quantitative measurement system for M2BPGi depends on liver fibrosis and inflammation, regardless of etiology. Liver inflammation complicates the interpretation of M2BPGi-Qt results when assessing the fibrosis stage.

Pancreatic cancer (PC) is a challenging malignancy to treat. Mac-2-binding protein glycan isomer (M2BPGi) is a novel serum marker of liver fibrosis and hepatocellular carcinoma and is secreted by hepatic stellate and stroma cells. Serum M2BPGi levels are upregulated in PC patients. We measured the expression of M2BPGi in the serum of 27 PC patients and determined whether M2BPGi affects the malignant potential of PC cells in vitro. We also examined the effect of M2BP on PC tumor growth and gemcitabine sensitivity in vivo. Serum M2BPGi levels in PC patients were higher compared with those of healthy subjects. M2BPGi extraction in cancer-associated fibroblasts (CAFs) was higher compared with that of PC cells. M2BPGi treatment promoted the proliferation and invasion of PC cells. The suppression of galectin-3, which binds to M2BPGi, did not affect the proliferation-promoting effect of M2BPGi in PC cells. The suppression of M2BP reduced tumor growth and enhanced gemcitabine sensitivity in PC-bearing xenograft mice. CAF-derived M2BPGi promotes the proliferation and invasion of PC cells. Targeting M2BPGi may represent a new therapeutic strategy to circumvent refractory PC.

Background Caspase-1 is a crucial component in the inflammasome activation cascade. This study evaluated the potential of serum caspase-1 level as an inflammatory biomarker in patients with adult-onset Still’s disease (AOSD). Methods The study included 51 consecutive patients diagnosed with AOSD based on the Yamaguchi criteria, 66 patients with rheumatoid arthritis (RA) as disease control, and 36 healthy controls (HCs). Serum caspase-1 concentrations were measured using enzyme-linked immunosorbent assay. The serum 69 cytokine levels were analyzed using a multisuspension cytokine array in patients with AOSD, and a cluster analysis of each cytokine was performed to determine specific molecular networks. Results Patients with AOSD had significantly increased serum caspase-1 levels versus patients with RA (p < 0.001) and HCs (p < 0.001). Additionally, serum caspase-1 demonstrated significant positive correlations with AOSD disease activity score (Pouchot score, r = 0.59, p < 0.001) and serum ferritin (r = 0.54, p < 0.001). Furthermore, among patients with AOSD, significant correlations existed between serum caspase-1 and inflammatory cytokines, including interleukin-18. Immunoblot analysis detected the cleaved form of caspase-1 (p20) in the serum of untreated patients with AOSD, not in those from patients with inactive AOSD receiving immunosuppressive treatments. Conclusions Caspase-1 is a useful biomarker for AOSD diagnosis and monitoring. Caspase-1 activation could be correlated with the inflammatory component of AOSD, specifically through proinflammatory cytokine induction via inflammasome activation cascades.

This study aimed to identify biomarkers to distinguish adult-onset Still’s disease (AOSD) and to predict disease phenotypes. In total, 49 patients diagnosed with AOSD and 200 patients with common diseases (controls) were included in the analysis. The levels of 69 cytokines were analyzed using a multi-suspension cytokine array. Cytokine cluster analysis was performed to identify specific molecular networks. Furthermore, random forest analysis and logistic regression analysis were used to rank cytokines based on their importance and to determine specific biomarkers for identification of AOSD patients and phenotypes. Patients with AOSD demonstrated significantly higher macrophage migration inhibitory factor (MIF) and interleukin (IL)-12(p40) serum levels than controls and patients with rheumatoid arthritis. Serum levels of chemokine (C-C motif) ligand (CCL) 8 and CCL22 were significantly lower in AOSD patients with a polycyclic systemic disease phenotype and could be differentiated with high accuracy from the other phenotypes (cutoff value for CCL8 = 122.7 pg/mL, CCL22 = 593.3 pg/mL, sensitivity 66.7%, specificity 87.1%, area under the curve 0.843). Combined MIF and IL-12(p40) levels may represent a biomarker for differentiating patients with AOSD from those with other diseases. The chemokine profiles of AOSD with a polycyclic systemic disease phenotype may differ from other phenotypes.

The integrative multi-kingdom interaction of the gut microbiome in ulcerative colitis (UC) and Crohn’s disease (CD) remains underinvestigated. Here, we perform shotgun metagenomic sequencing of feces from patients with UC and CD, and healthy controls in the Japanese 4D cohort, profiling bacterial taxa, gene functions, and antibacterial genes, bacteriophages, and fungi. External metagenomic datasets from the US, Spain, the Netherlands, and China were analyzed to validate our multi-biome findings. We found that Enterococcus faecium and Bifidobacterium spp. were enriched in both diseases. Enriched Escherichia coli was characteristic of CD and was linked to numerous antibiotic resistance genes involved in efflux pumps and adherent-invasive Escherichia coli virulence factors. Virome changes correlated with shifts in the bacteriome, including increased abundances of phages encoding pathogenic genes. Saccharomyces paradoxus and Saccharomyces cerevisiae were enriched in UC and CD, respectively. Saccharomyces cerevisiae and Escherichia coli had negative associations with short-chain fatty acid (SCFA)-producing bacteria in CD. Multi-biome signatures and their interactions in UC and CD showed high similarities between Japan and other countries. Since bacteria, phages, and fungi formed multiple hubs of intra- or trans-kingdom networks with SCFA producers and pathobionts in UC and CD, an approach targeting the interaction network may hold therapeutic promise. Here, the authors perform a multi-biome analysis in ulcerative colitis and Crohn’s disease patients from the Japanese 4D cohort, identifying intra- and trans-kingdom interactions including bacteria, phages, and fungi, providing potential candidate therapeutic targets.

Regular monitoring of patients with a history of hepatitis C virus (HCV) infection is critical for the detection and management of hepatocellular carcinoma (HCC). Mac-2 binding protein glycosylation isomer (M2BPGi) has been used to monitor fibrosis progression and predict HCC. However, HCC prediction based on M2BPGi has not been optimized. Here, we identified HCC risk-related glycan signatures of M2BP using a newly developed automated bead array with multiplexed lectins. Among 955 patients with HCV who achieved sustained virological response following direct-acting antiviral treatment, we compared M2BP glycosylation from sera of 42 patients diagnosed with HCC during follow-up and 43 without HCC (control) by the lectin microarray. At the HCC observation point, we found significant differences in 17 lectins. Using an automated bead array with 12 of 17 lectins, a principal component analysis (PCA) biplot differentiated HCC from control, along the PC1 axis, explaining 75.2% of variance. Based on PC1, we generated a scoring formula for an HCC-related glycosylation signature on M2BP (M2BPgs-HCC), showing good diagnostic performance for HCC (p = 2.92 × 10−8, AUC = 0.829). This automated multilectin bead array improved the ability of M2BP to detect HCC, providing a candidate test for HCC surveillance in combination with other HCC markers.

Monitoring of hepatitis B virus (HBV)-DNA and HBV-DNA-guided preemptive therapy using nucleos(t)ide analogs (NAs) are recommended to prevent the development of hepatitis due to HBV reactivation after allogeneic hematopoietic stem cell transplantation (allo-HSCT) in recipients with resolved HBV infection. However, little is known about the appropriate duration of NA treatment and the effect of NA cessation on the recurrence of HBV reactivation. This study aimed to clarify the consequences of NA cessation in allo-HSCT recipients with resolved HBV infection who experienced HBV reactivation following transplantation. We retrospectively reviewed the clinical records of recipients with resolved HBV infection (hepatitis B surface antigen [HBsAg]-negative, anti-HBc-positive) before allo-HSCT who had been diagnosed with HBV reactivation (HBsAg-positive and/or HBV-DNA detectable) after allo-HSCT between January 2010 and December 2020. A total of 72 patients from 16 institutions were registered (median age, 60 years; age range, 27 to 73 years; 42 males and 30 females). The day of initial HBV reactivation ranged from day 10 to day 3034 after allo-HSCT (median, 513 days). Anti-HBs were lost in >80% of the patients at the time of HBV reactivation. All 72 patients received preemptive NAs, and no fatal HBV reactivation-related hepatitis was observed. HBV-DNA without hepatitis was continuously detected in 5 patients during the follow-up period. Administration of NAs was discontinued in 24 of 72 patients (33%) by physician decision. Second HBV reactivation occurred in 11 of the 24 patients (46%) in whom administration of NAs was discontinued. The duration of NA treatment did not differ significantly between patients with or without second HBV reactivation. The frequency of further HBV reactivation tended to be lower in patients with an anti-HBs titer of >10 mIU/mL at the time of NA cessation. Multiple reactivations of HBV after NA cessation was common in patients with HBV reactivation who underwent allo-HSCT despite the long duration of NAs. Careful monitoring of HBV-DNA is important even after the discontinuation of NAs in the case with HBV reactivation after allo-HSCT, because multiple reactivations could occur. Active immunization by HB vaccine might be effective for suppressing further HBV reactivation after cessation of NAs.

Clinical and biochemical features of hepatitis delta virus (HDV) infections in Mongolia remain largely unknown. We aimed to investigate the clinical characteristics of HDV patients in Mongolia using several markers. The 143 hepatitis B surface antigen (HBsAg)-positive patients were divided into 122 HDV-positive and 21 HDV-negative patients by HDV RNA positivity. Subgroup analysis was performed between hepatitis B e antigen (HBeAg)-positive and -negative HDV-positive patients. Liver function, quantitative HBsAg (qHBsAg), anti-HDV Immunoglobulin (Ig) M, Mac-2 binding protein glycosylation isomer (M2BPGi), hepatitis B virus (HBV) DNA level, and HDV RNA level were tested. HDV RNA was positive in 85.3% (122/143) of patients showing anti-HDV IgG. Liver disease activity was higher in HDV-positive patients than in HDV-negative patients. The HDV-positive group included a higher proportion of patients with high qHBsAg and M2BPGi levels (p < 0.001). The positivity rate for anti-HDV IgM was significantly higher in the HDV-positive group (p < 0.001). HDV RNA levels showed an inverse correlation with qHBsAg levels in HBeAg-positive-HDV-positive patients (r = -0.49, p = 0.034), and a positive correlation with qHBsAg levels in HBeAg-negative patients (r = 0.35, p < 0.001). Hepatitis B virus (HBV) DNA and HDV RNA levels did not show any correlation. M2BPGi levels likewise did not correlate with HDV RNA levels. A high positivity rate for HDV RNA was observed for HBV patients in Mongolia using the highly sensitive HDV RNA assay. The positivity rate for anti-HDV IgM was high in HDV RNA-positive patients. Severity of liver disease and M2BPGi levels were both high in the HDV RNA-positive group.