Principais motivos de descarte de córneas para transplante na Paraíba: por que o anti-HBc merece atenção?* / Main reasons for discarding corneas for transplant in Paraíba: why does anti-HBc deserve attention?*

RESUMO Objetivo: Identificar o perfil dos doadores de tecidos oculares humanos na área de atuação do Banco de Olhos da Paraíba, destacando o impacto da sorologia positiva para hepatite B no descarte dos tecidos para transplante. Métodos: O estudo é transversal e utilizou dados do Banco de Olhos da Paraíba entre janeiro de 2013 e dezembro de 2022. Dados sobre procedência, idade, sexo, causa do óbito, tempo entre óbito e enucleação, resultados sorológicos e motivo de descarte das córneas dos doadores foram coletados. Resultados: O maior motivo de descarte foi por sorologia positiva (56,5%), sendo positivadas as sorologias positivas para hepatite B e HBsAg em 11,1% e 4,75% dos pacientes, respectivamente. Conclusão: A sorologia positiva para hepatite B como um critério de descarte absoluto é responsável por grande parcela de descartes, apesar da pouca informação sobre suas repercussões e representação de infectividade nos receptores do transplante.

ABSTRACT Objective: To identify the profile of human ocular tissue donors in the area covered by the Eye Bank of Paraíba (PB), highlighting the impact of positive serology for hepatitis B (anti-HBc) in the disposal of tissues for transplantation. Methods: This is a cross-sectional that uses data from the Eye Bank of Paraíba (PB) between January 2013 and December 2022. Data on origin, age, sex, cause of death, time between death and enucleation, serological results, and reason for discarded donor corneas were collected. Results: The main reason for discarding was due to positive serology (56.5%), with positive anti-HBc and HBsAg serology in 11.1% and 4.75% of patients, respectively. Conclusion: Anti-HBc positive serology as an absolute disposal criterion is responsible for great part of disposals, despite little information about its repercussions and representation of infectivity in transplant recipients.

EVALUATION OF SOME BIOCHEMICAL MARKERS AND THE ANTIBODY RESPONSE TO HEPATITIS B VACCINE IN HEMODIALYSIS PATIENTS.

OBJECTIVE: The aim: The goal of this study was to assess the immune response to the HB vaccine (the level of anti-HBs titer), as well as the prevalence of serum creatinine, urea, CRP, and serum albumin levels, and the relationship between these and immune response to the vaccine. PATIENTS AND METHODS: Materials and methods: 127 patients with chronic renal disease on hemodialysis (HD) were compared to 40 healthy people in Iraqi dialysis center, Baghdad. Antibodies to the hepatitis B core antigen (anti-HBc) were detected using the ARCHITECT SYSTEM and the Anti-HBs titer, HBs Ag, Anti-HCV determined by ELISA. RESULTS: Results: When compared to the poor and non-responder groups, the mean value of anti-HBs titer increased considerably in the good responder group. The good responder and control groups, on the other hand, showed no significant changes. The anti-HBs titer was found to have the strongest negative correlation with serum creatinine, blood urea, and C-reactive protein levels. There was a considerable positive connection between anti-HBs titer and albumin levels. CONCLUSION: Conclusions: The responses of HD patients to the HB vaccine revealed the significant negative relation between serum creatinine, blood urea levels, and CRP, as well as a significant positive correlation between serum albumin.

Incidence and prediction of HBsAg seroclearance in a prospective multi-ethnic HBeAg-negative chronic hepatitis B cohort.

BACKGROUND AND AIMS: Achieving HBsAg loss is an important landmark in the natural history of chronic hepatitis B (CHB). A more personalized approach to prediction of HBsAg loss is relevant in counseling patients. This study sought to develop and validate a prediction model for HBsAg loss based on quantitative HBsAg levels (qHBsAg) and other baseline characteristics. METHODS: The Hepatitis B Research Network (HBRN) is a prospective cohort including 1240 untreated HBeAg-negative patients (1150 adults, 90 children) with median follow-up of 5.5 years. Incidence rates of HBsAg loss and hepatitis B surface antibody (anti-HBs) acquisition were determined, and a predictor score of HBsAg loss using readily available variables was developed and externally validated. RESULTS: Crude incidence rates of HBsAg loss and anti-HBs acquisition were 1.6 and 1.1 per 100 person-years (PY); 67 achieved sustained HBsAg loss for an incidence rate of 1.2 per 100 PY. Increased HBsAg loss was significantly associated with older age, non-Asian race, HBV phenotype (inactive CHB vs. others), HBV genotype A, lower HBV-DNA levels, and lower and greater change in qHBsAg. The HBRN-SQuARe (sex,∆quantHBsAg, age, race) score predicted HBsAg loss over time with area under the receiver operating characteristic curve (AUROC) (95% CIs) at 1 and 3 years of 0.99 (95% CI: 0.987-1.00) and 0.95 (95% CI 0.91-1.00), respectively. In validation in another cohort of 1253 HBeAg-negative patients with median follow-up of 3.1 years, HBRN SQuARe predicted HBsAg loss at 1 and 3 years with AUROC values of 0.99 (0.98-1.00) and 0.88 (0.77-0.99), respectively. CONCLUSION: HBsAg loss in predominantly untreated patients with HBeAg-negative CHB can be accurately predicted over a 3-year horizon using a simple validated score (HBRN SQuARe). This prognostication tool can be used to support patient care and counseling.

Ubiquitous expression of HBsAg from integrated HBV DNA in patients with low viral load.

BACKGROUND & AIMS: Loss of serum HBsAg is a hallmark of spontaneous and therapy induced resolution of HBV infection, since it generally reflects a profound decrease in viral replication. However, integrated HBV DNA can contribute to HBsAg expression independent of viral replication. The relative contributions of these sources of HBsAg are not well understood. Specifically, it is not known whether actively transcribed HBV integration could spread throughout the entire liver. METHODS: The relative distribution of HBsAg and HBV RNA in liver biopsy tissue from HBeAg-negative (HBe-) patients was analyzed by immunohistochemistry and in situ hybridization (ISH), respectively. Frozen biopsy tissue was used for molecular analysis of intrahepatic viral RNA, virus-host chimeric transcripts and viral DNA. RESULTS: Immunohistochemistry and ISH analysis revealed HBsAg and HBV RNA positivity in virtually all hepatocytes in the liver of some HBe- patients despite very low viremia. Reverse transcription quantitative PCR and RNA-sequencing analysis confirmed high expression levels of HBV envelope-encoding RNAs. However, the amount of viral transcriptional template (covalently closed circular (ccc)DNA) was too low to support this ubiquitous HBV RNA expression. In contrast, levels of total cellular HBV DNA were consistent with ubiquitous HBV integration. Finally, RNA-sequencing revealed the presence of many HBV-host chimeric transcripts with the potential for HBsAg expression. CONCLUSIONS: Transcriptionally active HBV integration can extend to the entire liver in some HBe- patients. This can lead to ubiquitous HBsAg expression independent of HBV replication. In such patients, HBsAg is probably not a clinically useful surrogate marker for viral resolution or functional cure. LAY SUMMARY: Loss of serum hepatitis B surface antigen (HBsAg) indicates resolution of HBV infection. However, integrated HBV DNA can contribute to HBsAg production independently of viral replication. We investigated the extent of HBsAg-producing viral integration in the livers of patients with low serum viral loads. Our findings suggest that transcriptionally active HBV integration can extend to the entire liver in some patients, questioning the clinical utility of HBsAg as a surrogate marker for viral replication.

Hepatitis B virus vaccine and chronic kidney disease. The advances / Vacuna contra el virus de la hepatitis B y la nefropatía crónica: avances

BACKGROUND: Hepatitis B is an important agent of liver disease in patients with chronic kidney disease and chronic HBV infection promotes the development of CKD in the adult general population. Patients with CKD have a suboptimal response to various vaccines, and it remains unclear how we boost the immune response of CKD patients to HB vaccine. Study aims and design: We performed a narrative review to assess the mechanisms of lower immunogenicity of HBV vaccine in CKD population; multiple approaches to improve the response rate of CKD patients to HBV vaccine have been reported. This is a very important topic for nephrologists who often serve as primary case providers for patients with CKD. RESULTS: The recommended vaccine schedule for CKD patients including those on maintenance dialysis is based on recombinant vaccine, four doses (month 0,1,2, and 6; 40 mcg each) by intramuscular route (deltoid muscle). According to RCTs or observational studies, some recombinant vaccines with adjuvants (i.e., HBV-AS02 and HBV-AS04) look promising. HBV-AS04 showed to give better seroprotection rates and durable immune response over extended follow-ups compared with licensed HBV vaccine in CKD patients. The seroprotection rate was 95% (97/102) and 82% (202/248) in pre-dialysis and dialysis patients, respectively, one month after completing vaccine schedule with HBV-AS04. HBV-AS02 was superior to licensed vaccine in terms of seroprotection rate, 76.9% vs. 37.6%. CONCLUSIONS: We suggest adjuvanted recombinant (HBV-AS04) vaccine (0,1,2 and 3 months; 20 mcg each dose) and post vaccination testing of anti-HBs antibody after vaccination. Booster doses to patients whose anti-HBs titers fall below the seroprotection level (< 10 IU/mL) during the follow-up are appropriate. The patho-physiologic mechanisms responsible for the poor immunogenicity of HBV vaccine in CKD patients are under active investigation

ANTECEDENTES: La hepatitis B es un importante agente de la enfermedad hepática en pacientes con nefropatía crónica (NC) y la infección crónica por el virus de la hepatitis B (VHB), promueve el desarrollo de la NC en la población general adulta. Los pacientes con NC tienen una respuesta subóptima a varias vacunas, y no está claro cómo podemos aumentar la respuesta inmunológica de estos pacientes a la vacuna contra el VHB. Objetivos y diseño del estudio: Realizamos una revisión narrativa para evaluar los mecanismos de menor inmunogenicidad de la vacuna contra el VHB en la población con NC; se han documentado varios enfoques para mejorar la tasa de respuesta de los pacientes con NC a la vacuna contra el VHB. Este es un tema muy importante para los nefrólogos, que a menudo atienden como médicos de atención primaria a pacientes con NC. RESULTADOS: El programa de vacunación recomendado para los pacientes con NC, incluidos los que están en diálisis de mantenimiento, se basa en una vacuna recombinante de cuatro dosis (meses 0, 1, 2 y 6; 40 mcg cada dosis), administrada por vía intramuscular (músculo deltoides). Según los ECA, o estudios observacionales, algunas vacunas recombinantes con adyuvantes (es decir, HBV-AS02 y HBV-AS04) parecen prometedoras. La HBV-AS04 demostró ofrecer mejores tasas de seroprotección y una respuesta inmunitaria duradera durante los seguimientos prolongados, en comparación con la vacuna autorizada contra el VHB en pacientes con NC. La tasa de seroprotección fue del 95% (97/102) y del 82% (202/248), en pacientes en prediálisis y diálisis, respectivamente, un mes después de completar el programa de vacunación con HBV-AS04. La HBV-AS02 fue superior a la vacuna autorizada con respecto a la tasa de seroprotección, 76,9 vs. a 37,6%. CONCLUSIONES: Sugerimos una vacuna recombinante con adyuvante (HBV-AS04) (meses 0, 1, 2 y 3, 20 mcg cada dosis) y pruebas de anticuerpos anti-HB después de la vacunación. Se consideran adecuadas las dosis de refuerzo para los pacientes cuyos títulos de anticuerpos anti-HB sean inferiores al nivel de seroprotección (< 10 UI/mL) durante el seguimiento. Los mecanismos fisiopatológicos responsables de una inmunogenicidad deficiente de la vacuna contra el VHB en pacientes con NC son objeto de investigaciones exhaustivas

Clinical efficacy of a novel, high-sensitivity HBcrAg assay in the management of chronic hepatitis B and HBV reactivation.

BACKGROUND & AIMS: A fully automated, novel high-sensitivity hepatitis B core-related antigen assay (iTACT-HBcrAg) has been developed. We demonstrate the clinical utility of iTACT-HBcrAg for monitoring chronic hepatitis B (CHB) and for the early detection of HBV reactivation. METHODS: After fundamental assessments, the clinical performance of iTACT-HBcrAg was compared with other HBV markers. i) Serial sera, available from 161 HBeAg-negative patients with CHB and persistently undetectable HBV DNA, were measured by iTACT-HBcrAg and a conventional HBcrAg assay (G-HBcrAg). ii) Serial sera from 13 HBV-reactivated patients were measured by iTACT-HBcrAg and an ultra-high-sensitivity HBsAg immune complex transfer-chemiluminescent enzyme immunoassay (lower limit of detection; 0.0005 IU/ml, ICT-CLEIA) to compare HBV DNA detection. iii) To elucidate the various HBcrAg components detected by iTACT-HBcrAg, OptiPrep density gradient centrifugation analysis was performed on sera obtained before and after HBV reactivation. RESULTS: The analytical performance of iTACT-HBcrAg was satisfactory. The sensitivity of iTACT-HBcrAg (2.1 Log U/ml) was approximately 10-fold greater than that of G-HBcrAg (2.8 Log U/ml). i) HBcrAg was detectable in the sera of 97.5% (157/161) of patients with CHB by iTACT-HBcrAg, of whom 75.2% (121/161) had ≥2.8 Log U/ml HBcrAg and 22.4% (36/161) had 2.1-2.8 Log U/ml HBcrAg, which was undetectable by G-HBcrAg. ii) 9 and 2 of 13 HBV-reactivated patients were HBcrAg-positive by iTACT-HBcrAg before and at HBV DNA positivity, respectively; 7 and 4 were HBcrAg-positive by iTACT-HBcrAg before and at HBsAg-positivity by ICT-CLEIA, respectively. iii) The HBcrAg detected by iTACT-HBcrAg before HBV reactivation was contained in empty particles (22 KDa precore protein). CONCLUSIONS: iTACT-HBcrAg could be used to better monitor responses to anti-HBV treatments in HBeAg-negative patients and for the early detection of HBV reactivation. LAY SUMMARY: A fully automated, novel high-sensitivity hepatitis B core-related antigen assay (iTACT-HBcrAg) has been developed. iTACT-HBcrAg can be used to monitor HBeAg-negative patients with chronic hepatitis B, as well as for the early detection of HBV reactivation. iTACT-HBcrAg could be used as a general marker of disease progression and treatment response.

Risk of hepatitis B reactivation in patients receiving anti-tumor necrosis factor-α therapy.

OBJECTIVE: The purpose of this study was to determine hepatitis B virus (HBV) screening rates in patients receiving anti-tumor necrosis factor (TNF)-α therapy and the frequency of HBV reactivation in patients with resolved hepatitis B virus infection (hepatitis B surface antigen [HBsAg] negative, hepatitis B core antibody [Anti-HBc] positive). PATIENTS AND METHODS: Data from 1834 patients who underwent anti-TNF-α therapy in the Rheumatology, Gastroenterology and Dermatology Departments of our hospital between 2010 and 2020 were retrospectively analyzed. Within 6 months before the initial anti-TNF-α therapy, performing a HBsAg and/or anti-HBc test is defined as HBV screening. HBV reactivation is defined as the presence of detectable serum HBV DNA or HBsAg seroconversion from negative to positive. RESULTS: The overall HBV screening rate was 82.3% before starting anti-TNF-α therapy. There was an increasing trend in HBV screening rates during the years analyzed (64% in 2010, 87.4% in 2019) (P < .001). Before anti-TNF-α therapy was initiated, 272 patients were HBsAg negative and anti-HBc positive. Among these patients, HBV reactivation did not occur in 31 patients who received antiviral prophylaxis, whereas HBV reactivation occurred in only 1 (0.4%) of the 241 patients who did not receive antiviral prophylaxis. CONCLUSION: Hepatitis B virus screening rates prior to starting anti-TNF-α therapy were relatively high, and its trend was increased by year. HBV reactivation because of anti-TNF-α use rarely occurred in patients with resolved HBV infection. Further studies are needed on whether routine anti-HBc screening and/or HBV DNA follow-up are necessary in these patients aside from HBsAg.

Specific determination of hepatitis B e antigen by antibodies targeting precore unique epitope facilitates clinical diagnosis and drug evaluation against hepatitis B virus infection.

Hepatitis B e antigen (HBeAg) is a widely used marker both for chronic hepatitis B (CHB) clinical management and HBV-related basic research. However, due to its high amino acid sequence homology to hepatitis B core antigen (HBcAg), most of available anti-HBe antibodies are cross-reactive with HBcAg resulting in high interference against accurate measurement of the status and level of HBeAg. In the study, we generated several monoclonal antibodies (mAbs) targeting various epitopes on HBeAg and HBcAg. Among these mAbs, a novel mAb 16D9, which recognizes the SKLCLG (aa -10 to -5) motif on the N-terminal residues of HBeAg that is absent on HBcAg, exhibited excellent detection sensitivity and specificity in pairing with another 14A7 mAb targeting the HBeAg C-terminus (STLPETTVVRRRGR, aa141 to 154). Based on these two mAbs, we developed a novel chemiluminescent HBeAg immunoassay (NTR-HBeAg) which could detect HBeAg derived from various HBV genotypes. In contrast to widely used commercial assays, the NTR-HBeAg completely eliminated the cross-reactivity with secreted HBcAg from precore mutant (G1896A) virus in either cell culture or patient sera. The improved specificity of the NTR-HBeAg assay enables its applicability in cccDNA-targeting drug screening in cell culture systems and also provides an accurate tool for clinical HBeAg detection.

Development of a multiplex immunochromatographic strip test and ultrasensitive electrochemical immunosensor for hepatitis B virus screening.

This research proposes two methods for hepatitis B diagnosis including rapid testing and electrochemical assay. For the first method, a multiplex hepatitis B test strip was fabricated to serve as a rapid test for hepatitis B screening. It was developed to simultaneously test three essential serological markers of hepatitis B virus infection including hepatitis B surface antigen (HBsAg), hepatitis B surface antibody (Anti-HBs) and hepatitis B core antibody (Anti-HBc). Gold nanoparticles (GNPs) were used as the signal generator on the test strip. Furthermore, a part of a paper network was incorporated on the strip for the gold-silver enhancement process. This paper network helped in decreasing the analysis time of enhancement and makes the enhancement process easier for rapid testing. The developed test strip was specific for each serological marker. The detection limits of HBsAg, Anti-HBs and Anti-HBc were obtained at 0.5, 0.3 and 0.1 µg mL-1, respectively. For the second method, electrochemical impedance spectroscopy (EIS) was applied for HBsAg detection. This method was proposed for quantitative hepatitis B detection. Anti-HBs antibodies were immobilized on a carbon screen printed electrode (SPCE) via the N-ethyl-N'-(3-(dimethylamino)propyl)carbo-diimide/N-hydroxy succinimide (EDC/NHS) couple reaction which reacted with the carboxyl group of the BSA cross-linked film on the electrode. The electrode modification process was characterized by EIS. A linear relationship between delta charge transfer resistance (ΔRct) and HBsAg concentration was obtained in the range of 5-3000 ng mL-1 with a detection limit of 2.1 ng mL-1. This work is appropriate for quantitative analysis because it is a simple and low-cost method to implement as the SPCE is disposable. Therefore, we hope that this research will be useful to improve hepatitis B detection in the future.

Risk stratification and clinical course of hepatitis B virus reactivation in rheumatoid arthritis patients with resolved infection: final report of a multicenter prospective observational study at Japanese Red Cross Hospital.

BACKGROUND: The prophylaxis for hepatitis B virus (HBV) reactivation assumes that hepatic injury after reactivation is often rapidly progressive and can evoke fulminant hepatitis. The incidence and prognosis of reactivation in patients with rheumatoid arthritis (RA) may be different from those receiving organ transplantation and cancer chemotherapy. This study aimed to investigate the incidence, risk factors, and clinical course of HBV reactivation and develop a scoring system for risk stratification in RA patients with resolved infection. METHODS: HBV DNA was measured using real-time polymerase chain reaction, and patient data were collected for 4 years in RA patients with resolved HBV infection who were treated with steroids or synthetic or biologic immunosuppressive drugs. RESULTS: Among 1127 patients, HBV DNA was detected in 57 patients (1.65/100 person-years); none of the reactivated patients exhibited worsening of hepatic function. Multivariate logistical analysis revealed that age > 70 years and HB core antibody (HBcAb) positivity alone were independent risk factors for HBV reactivation. HBV DNA ≥ 2.1 log copies/mL was observed in 15 patients (0.43/100 person-years); seven patients were treated with nucleic acid analogs (NAAs), whereas the remaining eight were observed without treatment. Among reactivated cases, 15 cases changed to HBV DNA-negative status spontaneously, whereas 24 cases remained HBV DNA positive < 2.1 log copies/mL during the observation period. We designed the following scoring system: HBV reactivation risk score = 1 × (age > 70 years) + 2 × (HBcAb positivity alone) + 1 × (treatment other than methotrexate monotherapy). This revealed that patients with the highest score had an odds ratio of 13.01 for HBV reactivation, compared to those with the lowest score. CONCLUSIONS: Rapid progression and poor outcomes after HBV reactivation were not frequent in RA patients with resolved infection. Our new risk scoring system might be useful for screening and optimization of prophylactic treatment by distinguishing patients with significantly lower reactivation risk.

Determination of Hepatitis B Immunoglobulin Infusion Interval Using Pharmacokinetic Half-life Simulation for Posttransplant Hepatitis B Prophylaxis.

BACKGROUND: Prophylaxis for hepatitis B virus (HBV) recurrence is essential after liver transplantation (LT) in HBV-associated recipients. This study established an individualized HBV prophylaxis protocol, through optimization of hepatitis B immunoglobulin (HBIG) administration, with application of simulative half-life (SHL). METHODS: This study involved five parts: Part 1 developed the SHL estimation method with 20 patients; Parts 2 and 3 assessed the SHL variability and developed a simulation model to apply SHL in 100 patients; Part 4 validated the simulation model in 114 patients, and Part 5 was a cross-sectional study on the current status of HBIG infusion intervals in 660 patients. RESULTS: In Part 1, infusion of 10,000 IU HBIG induced add-on rise hepatitis B surface antibody (anti-HBs) titer of 5,252.5 ± 873.7 IU/L, which was 4.4% lower than actual measurement. Mean SHL of 20.0 ± 3.7 days was 2.2% longer than actual measurement. In Part 2, the medians of the intra- and inter-individual coefficient of variation in SHL were 13.5% and 18.5%, respectively. Pretransplant HBV DNA load and posttransplant antiviral therapy did not affect SHL. In Part 3, a simulation model was developed to determine the interval of HBIG infusion, by using SHL. In Part 4, all 114 patients were successfully managed with regular HBIG infusion intervals of ≥ 8 weeks, and the interval was prolonged to ≥ 12 weeks in 89.4%, with a target trough anti-HBs titer ≥ 200 IU/L. In Part 5, 47.4% of our patients received HBIG excessively, at a target trough titer of 500 IU/L. CONCLUSION: SHL estimation using only clinically available parameters seems to be reliably accurate when compared with actual measurements. We believe that SHL estimation is helpful to establish a personalized HBV prophylaxis protocol for optimizing HBIG administration.

Evaluation of occult hepatitis B infection in tissue donors: a multicenter analysis in Spain.

Traditionally, when antibody to the Hepatitis B core antigen (anti-HBc) and antibody to the Hepatitis B surface antigen (anti-HBs) are positive, the donor is considered suitable. However, the literature contains cases with this profile and circulating hepatitis B virus DNA. The aim of the study is to analyze the incidence of occult hepatitis B virus infection (OBI). Retrospective data were evaluated for deceased tissue donors in ten Tissue Establishments (Spain) during 2017. The data included demographic data and the serological markers for hepatitis B that each tissue establishment performed. A total number of 1933 tissue donors were evaluated. A total of 180 donors were excluded: 6 (0.3%) with Hepatitis B surface antigen (HBs positive), and 174 in which DNA testing was not performed. Anti-HBc was positive in 175 donors (10%), in which anti-HBs was negative in 30 (17.1%) and positive in 145 (82.9%). In total, 27 donors with DNA positive (1.5%) were found, of which 3 of 117 donors (1.7%) showed anti-HBc negative and anti-HBs positive (> 10 IU/ml), 4 of 30 donors (13.3%) showed anti-HBc positive and anti-HBs negative and 20 of 145 donors (13.8%) showed both anti-HBc and anti-HBs positive. The highest probability of finding DNA occurs when anti-HBc is positive, regardless of the presence of anti-HBs. In our study, the probability of OBI was 1.5%. The classic concept that when anti-HBc and anti-HBs are positive (even with a titer of over 100 IU/ml) the donor can be accepted should, therefore, be reconsidered, and DNA testing should be mandatory.

Utility of oral fluid samples for hepatitis B antibody detection in real life conditions.

BACKGROUND: Hepatitis B virus (HBV) testing in oral fluid samples may provide advantages in diagnosis, screening or prevalence studies, especially among individuals with venous access difficulties. This study aims to optimize one commercially available assay for detecting total anti-HBc marker in oral fluid samples and to evaluate its utility under real life conditions in different settings for the purposes of prevalence and diagnostic studies. METHODS: Oral fluid was collected using a Salivette device and some parameters were initially evaluated: type of elution buffer and sample volume. Thereafter, the utility of oral fluid samples for detection of anti-HBc was evaluated in real life conditions in which, 1296 individuals gave serum and oral fluid samples. All serum samples were submitted to commercial EIAs to detect total anti-HBc, according to the manufacturer's instructions and oral fluid samples according to previous optimization. RESULTS: In optimization evaluation, PBS/BSA 0.5% and 100 µL of oral fluid (volume was two-fold increased compared to serum in EIA) were chosen as transport buffer and sample volume. In the field study, anti-HBc was detected in 211 out of 1296 serum samples giving overall oral fluid sensitivity of 52.6% and specificity of 96%. Concordance was higher in ambulatory setting (67.7) compared to general population (31.8). Mean ± standard deviation values of optical density/cutoff (OD/CO) in serum samples were higher in false-negative oral fluid samples than those seen in true positive samples. Sensitivity was higher in those presenting active infection compared to anti-HBc isolate and past infection. Sensitivity also increased in the ambulatory group when HCV individuals were excluded. CONCLUSIONS: It was possible to optimize a commercial EIA for detecting anti-HBc in oral fluid samples and where the highest concordance was found in ambulatory settings and among individuals with active infection.

Hepatitis B virus vaccination and revaccination response in children diagnosed with coeliac disease : a multicentre prospective study.

AIM: This study evaluates hepatitis B virus (HBV) vaccination response in children with celiac disease (CD). Response in initial non-responders after a single booster vaccination as well as factors influencing HBV vaccination response were evaluated. METHODOLOGY: Anti-hepatitis B surface antibodies (a-HBsAB) were checked in all children with CD and a documented complete HBV vaccination. An a-HBsAB <10 U/L was considered as non-response. A single intramuscular HBV-vaccine booster was advised to all non-responders. Response was checked at the next appointment. RESULTS: 133 children with CD were included, median age of 7.3 years (range 1.7-17.3) and 46 (35%) were male. The age at CD diagnosis was 6.0 years (range 1.1-15.7). HBV non-response was documented in 55% (n=73/133). No other factors were influencing the response. A booster was documented in 34/73 (47 %) initial non-responders (3 refused (4%), 36 (49%) had no follow up). Response after booster vaccination resulted in immunity in 22/34 (65%) and persisting non-response in 12/34 (35%). A single booster is able to reduce non-response from 55% (73/133) to 23% (22/94). CONCLUSION: A significantly lower immune response following HBV vaccination in children with CD was confirmed. A single intramuscular booster vaccination is able to induce a serologic response in two thirds of the initial non-responders. Control of HBV vaccination response has to become part of the follow-up in CD patients.

Liver transplantation using hepatitis B core positive grafts with antiviral monotherapy prophylaxis.

BACKGROUND & AIMS: The impact of hepatitis B core antibody (anti-HBc) positive liver grafts on survival and the risk of de novo hepatitis B virus (HBV) infection after liver transplantation (LT) remain controversial. Therefore, we aimed to analyze this risk and the associated outcomes in a large cohort of patients. METHODS: This was a retrospective study that included all adults who underwent LT at Queen Mary Hospital, Hong Kong, between 2000 and 2015. Data were retrieved from a prospectively collected database. Antiviral monotherapy prophylaxis was given for patients receiving grafts from anti-HBc positive donors. RESULTS: A total of 964 LTs were performed during the study period, with 416 (43.2%) anti-HBc positive and 548 (56.8%) anti-HBc negative donors. The median follow-up time was 7.8 years. Perioperative outcomes (hospital mortality, complications, primary nonfunction and delayed graft function) were similar between the 2 groups. The 1-, 5- and 10-year graft survival rates were comparable in anti-HBc positive (93.3%, 85.3% and 76.8%) and anti-HBc negative groups (92.5%, 82.9% and 78.4%, p = 0.944). The 1-, 5- and 10-year patient survival rates in anti-HBc positive group were 94.2%, 87% and 79% and were similar to the anti-HBc negative group (93.5%, 84% and 79.7%, p = 0.712). One-hundred and eight HBsAg negative recipients received anti-HBc positive grafts, of whom 64 received lamivudine and 44 entecavir monotherapy prophylaxis. The risk of de novo HBV was 3/108 (2.8%) and all occurred in the lamivudine era. There were 659 HBsAg-positive patients and 308 (46.7%) received anti-HBc positive grafts. The risk of HBV recurrence was similar between the 2 groups. Donor anti-HBc status did not impact on long-term patient and graft survival, or the risk of hepatocellular carcinoma recurrence after LT. CONCLUSIONS: De novo HBV was exceedingly rare especially with entecavir prophylaxis. Anti-HBc positive grafts did not impact on perioperative and long-term outcomes after transplant. LAY SUMMARY: The risk of de novo hepatitis B infection after liver transplantation was rare when using hepatitis B core positive liver grafts with entecavir monotherapy prophylaxis. Hepatitis B core antibody status did not impact on perioperative and long-term outcomes after liver transplantation. This provides support for the clinical use of hepatitis B core positive liver grafts when required.

Monitoring and Treatment of Patients Undergoing Immunotherapy With Anti-CD20 Who are Exposed to HBV.

Reactivation of hepatitis B virus (HBV) is a potentially fatal complication of immunosuppressive therapy, and can occur in individuals who are hepatitis B surface antigen (HBsAg) negative but positive for hepatitis B core antibody (anti-HBc). While anti-HBc positivity indicates prior HBV exposure, it may also reflect clearance of HBsAg, but with viral persistence at low intrahepatic replicative and transcriptional levels.1 HBV reactivation can still occur during intense immunosuppression, including B cell-depleting therapy with anti-CD20 antibodies2 and hematopoietic stem cell transplantation.3 While prevention via antiviral prophylaxis is recommended, it remains uncertain, from a global perspective, if this is an ideal and cost-effective strategy. An alternative is regular HBV DNA monitoring.4 However, this approach is problematic in resource-constrained regions, where the logistics of sample collection, transportation, and molecular analysis in dedicated facilities poses challenges.5 We aimed to evaluate the effectiveness of simple monitoring strategies using routine liver biochemistry and serum HBsAg in preventing HBV-related complications during anti-CD20 therapy.

Persistence of hepatitis B surface antibody and immune memory to hepatitis B vaccine among medical college students in Madinah.

BACKGROUND: Globally, about 300 million people are infected with hepatitis B virus (HBV). Among the effective approaches to fight HBV infection is immunization. In 1989, an obligatory hepatitis B vaccine program was launched in Saudi Arabia. OBJECTIVE: Assess hepatitis B surface antibody (anti-HBs) levels among the medical students before and after receiving booster doses of HBV vaccine. DESIGN: Cross-sectional. SETTING: Taibah University. SUBJECTS AND METHODS: Students born between 1993 and 1995 were recruited in this study from the Occupational Health Clinic. Students were screened for anti-HBs levels using chemiluminescent microparticle immunoassay (CMIA) before and after booster HBV vaccine doses. MAIN OUTCOME MEASURES: Anti-HBs levels before and after booster doses. SAMPLE SIZE: 335. RESULTS: About half of participants (n=164, 49%) had protective anti-HBs levels ( greater than or equal 10 mIU/mL) to the original primary series of HBV vaccine and received no booster doses. The reimaining 171 (51%) participants were at risk of HBV infection since their anti-HBs levels were less than 10 mIU/mL, despite having received the original primary HBV vaccine. The levels of anti-HBs were higher in female than in male students (P less than .001). In addition, female students showed a stronger humoral immune response to the booster vaccine than male students (P less than .001). When participants were given the three boosters, most participants (98.3%) showed anti-HBs levels of greater than or equal 10 mIU/mL. The results also showed a strong correlation between pre-booster and post-booster anti-HBs levels in the greater than or equal 10 mIU/mL group (r2= 0.52, P less than .001) but not in less than 10 mIU/mL group (r2= 0.003, P=.53). CONCLUSION: A considerable portion of the participants (about 51%) were at risk of HBV infection since their anti-HBs levels were less than 10 mIU/mL. Booster doses significantly trigger memory immune response and this ensured their protection against the virus. Pre-booster anti-HBs level are a good predictive of post-booster anti-HBs levels in greater than or equal 10 mIU/mL group. LIMITATIONS: The sample size was small. Shortage of collaborators. CONFLICT OF INTEREST: None.

Progression from acute to chronic hepatitis B is more common in older adults.

PURPOSE: The rate of progression of acute Hepatitis B (HBV) to chronic disease is quoted as <10%. The purpose of this study was to determine the rate of progression from acute to chronic HBV in Northern Ireland (NI), assessing the influence of age, gender and biochemical parameters. METHODS: All "acute" HBV cases diagnosed in NI between 2011 and 2015 were reviewed. Inclusion criteria: 1). positive HBsAg and positive HBV core IgM; 2). in the absence of positive HBV core IgM, positive HBsAg with a recent negative HBsAg. Patient age, HBsAg, HBV core IgM, peak bilirubin and peak ALT were recorded, along with date and result of repeat HbsAg testing. Mann-Whitney U test was used to compare mean age, peak ALT and bilirubin between clearing and non-clearing groups. Fisher's exact test was used to compare progression to chronicity according to gender and age less than or greater than 50yrs. RESULTS: Of 80 identified cases, 4 incorrectly categorised cases were excluded. Of the remaining 76, (15 female (mean age 37.27yr), 61 male (mean age 47.39yr)) follow-up data was available for 71 patients (15 female (mean age 37.27yr), 56 male (48.59yr)). All female patients cleared HBV. 42 of 61 males cleared HBV (p=0.0313).Overall the chronicity rate was 18.42% The mean age of those clearing the virus was 43.88 years, versus 55.64 years for those going on to develop chronic HBV (Mann-Whitney U test, z= -2.68, p=0.0037). Clearance rate was 83.72% in patients aged <50yrs and 63.64% in patients 50yrs (p=0.0068).Mean peak ALT (U/L) and peak bilirubin (µmol/L) for the clearing group were 2130 and 174 respectively compared to 656 and 100 for the non-clearing group (z= -3.51, p=0.0002, z= -2.35, p=0.009). CONCLUSION: Our results suggest a higher than expected rate of progression from acute to chronic HBV with a significantly higher risk for those over 50yrs. This suggests a need to revise information provided to older patients with acute HBV regarding the likelihood of progression.

Determination of hepatitis B, C and D prevalence among urban and Amerindian populations from the Eastern Brazilian Amazon: a cross sectional study.

BACKGROUND: This study was conducted to determine the prevalence of HBV, HCV, and HDV in urban populations and Amerindians living in the state of Tocantins (Eastern Amazon). METHODS: A total of 948 individuals were recruited in Tocantinopolis city (Tocantins state) of whom 603 were Amerindians (from 6 tribes) and 345 were non-Amerindians (6 urban areas of Tocantinópolis city). Anti-HCV, HBsAg, anti-HBc, anti-HBs, anti-HBc IgM, anti-HBe, HBeAg, and anti-delta antibodies were determined using enzyme immunoassay. RESULTS: HBV cleared infection (both anti-HBc/anti-HBs+), chronic inactive/immune controlled HBV infection (anti-HBc + only), previous HBV vaccination (anti-HBs + only), active HBV infection (HBsAg+), individuals susceptible to HBV, and anti-HCV reactivity were found in 12.9, 1.8, 27.2, 0.5, 57.7, 1.2% in Amerindians and 12.1, 2.0, 37.1, 0.3, 55.4, 0.3% in non-Amerindians respectively. Out of 139 anti-HBc reactive individuals, 70 were anti-HBe reactive and none presented HBeAg or anti-HBc IgM. Anti-HBc prevalence was associated to older age (p < 0.0001). Overall anti-Delta prevalence was 0.3% and regarding anti-HBc reactive individuals, anti-delta prevalence was 3.4 and 0% in Amerindians and non-Amerindians respectively. CONCLUSIONS: Overall low prevalence of HBV and HCV infection was found in the populations studied, but high HBV and HCV prevalence was observed in Amerindians compared to non-Amerindians suggesting that these individuals have a higher likelihood of acquiring to these infections. Anti-delta antibodies were found among Amerindians from Eastern Amazon suggesting a risk for this population. Of note is that nearly half of Amerindians had no anti-HBs, indicating a need for HBV vaccination campaigns in this population.