Prophylaxis and treatment in liver transplantation. VII Consensus Document of the Spanish Society of Liver Transplantation. / Profilaxis y tratamiento de la infección por virus de la hepatitis B en el trasplante hepático. VII Documento de consenso de la Sociedad Española de Trasplante Hepático.

Whilst prophylaxis of hepatitis B is universally accepted after liver transplantation (LT), national recommendations for the prophylaxis and treatment of hepatitis B virus (HBV) infection after LT are lacking in Spain. The aim of the VII consensus meeting organised by the Spanish Society of Liver Transplantation (SETH) was to set recommendations on the prophylaxis and treatment of hepatitis B after LT. The scientific evidence and strength of recommendations was evaluated by using the "Grading of Recommendations Assessment, Development and Evaluation" (GRADE) system. This document describes the recommendations and their level of evidence for: the definition and risk factors for hepatitis B recurrence after LT, monitoring and prophylaxis of hepatitis B recurrence at different periods after LT, treatment of hepatitis B before and after LT, and the prophylaxis of HBV infection by the recipients of LT with hepatitis B core antigen positive donors.

Serología del virus de la hepatitis B: para múltiples escenarios, múltiples exámenes / Serology of hepatitis B virus: multiple scenarios and multiple exams

Resumen El virus de la hepatitis B (VHB) tiene un gran impacto mundial. No obstante la disponibilidad de la vacuna, 2000 millones de personas se han infectado agudamente y, de ellos, 240 millones persisten crónicamente infectados. La infección tiene diferentes formas de presentación tales como infección aguda, infección crónica, infección oculta y reactivación cuando hay inmunosupresión. Así mismo, hay marcadores muy sensibles como el anticore, cuya positividad puede tener diversos significados. El recientemente descrito antígeno relacionado con el antígeno core es un marcador emergente que podría reemplazar al ácido desoxirribonucleico (ADN) viral. En la presente revisión se discuten los exámenes de laboratorio necesarios para el diagnóstico de los diferentes escenarios de la infección.

Abstract Hepatitis B virus (HBV) has an enormous global impact. Despite the availability of a vaccine, two billion people have been acutely infected. Of these, 240 million remain chronically infected. The infection has different forms of presentation including acute infections, chronic infections, hidden infections, and reactivation when there is immunosuppression. Similarly, there are very sensitive markers such as anti-core, but a positive test can have different meanings. This recently described antigen which is related to the core antigen is an emerging marker that could replace viral DNA. In this review we discuss the laboratory tests necessary for diagnosing the various scenarios of the infection.

Isolation and Characterization of Bluetongue Virus Recovered from Blood Samples by Immunoaffinity Purification.

An immunoaffinity chromatography (IAC) method was optimized for the selective capture of bluetongue virus (BTV) from blood samples and isolation of the virus in cell culture. The antibody against BTV core particles (lacking the outer capsid proteins VP2 and VP5) was used for the optimization of IAC technique. The antibody against BTV core particle was conjugated with Protein A-virus complex and the complex was dissociated using elution buffer (4 M MgCl2 with 75 mM HEPES, pH 6.5). The optimized IAC method specifically purified the BTV without capturing other commonly infecting small ruminant's viruses like gaotpox virus (GTPV), sheeppox virus (SPPV), Peste des petits ruminants virus (PPRV) and Foot and mouth disease virus (FMDV). The blood samples (n = 22), positive for BTV antigen in sandwich-ELISA were attempted for virus isolation in the BHK-21 cell using the optimized IAC method. A total of seven BTV were isolated by selective capturing of the virion particles. The isolated viruses were characterized by RNA-PAGE, sequence analysis and serum neutralization test (SNT). Electropherotypic analysis of viral dsRNA in the RNA-PAGE revealed the presence of ten dsRNA segments characteristic of BTV. Out of seven isolates, four isolates were identified as BTV-1 and three isolates were identified as BTV-16 based on nucleotide sequences of segment-2. Phylogenetic analysis of segment-2 nucleotide sequence segregated BTV-1 and BTV-16 isolates to monophyletic cluster at close proximity to other eastern topotype. In SNT, hyperimmune serum (HIS) against BTV-1 neutralized the four BTV-1 isolates up to a titer > 256 and HIS against BTV-16 neutralized the three BTV-16 isolates up to a titer > 128. The IAC technique will be useful for the selective capture of BTV from mixed infection (BTV with other small ruminant's viruses) and isolation from blood sample having low viral load by enrichment.

Quantification of HBV core antibodies may help predict HBV reactivation in patients with lymphoma and resolved HBV infection.

BACKGROUND & AIMS: Absence or low anti-HBV surface antibody (anti-HBs) is associated with an increased risk of HBV reactivation in patients with lymphoma and resolved HBV infection receiving rituximab-containing chemotherapy. Quantification of anti-HBV core antibody (anti-HBc) is a new marker associated with the natural history and treatment response of chronic HBV infection. This study investigated whether baseline anti-HBc and anti-HBs levels may better predict HBV reactivation. METHODS: We prospectively measured the HBV DNA levels of patients with lymphoma and resolved HBV infection receiving rituximab-cyclophosphamide, hydroxydaunorubicin, vincristine, and prednisolone-based chemotherapy and started an antiviral therapy upon HBV reactivation, defined as a greater than 10-fold increase in HBV DNA compared with previous nadir levels. Anti-HBs and anti-HBc were quantified by a double-sandwich assay. Receiver-operating-characteristic-curve analysis was used to determine the optimal baseline anti-HBc/anti-HBs levels for predicting HBV reactivation. RESULTS: HBV reactivation occurred in 24 of the 197 patients enrolled, with an incidence of 11.6/100 person-years. For the 192 patients with enough serum samples for analysis, low anti-HBs (<56.48 mIU/ml) and high anti-HBc (≥6.41 IU/ml) at baseline were significantly associated with high risk of HBV reactivation (hazard ratio [HR] 8.48 and 4.52, respectively; p <0.01). The multivariate analysis indicated that (1) patients with both high anti-HBc and low anti-HBs at baseline (36 of 192 patients) had an HR of 17.29 for HBV reactivation (95% CI 3.92-76.30; p <0.001), and (2) HBV reactivation may be associated with inferior overall survival (HR 2.41; 95% CI 1.15-5.05; p = 0.02). CONCLUSIONS: Baseline anti-HBc/anti-HBs levels may predict HBV reactivation in these patients with lymphoma and help optimize prophylactic antiviral therapy for high-risk patients. LAY SUMMARY: In this study, we identified a subgroup of patients with lymphoma and resolved hepatitis B virus infection that had a high risk of hepatitis B virus reactivation after receiving rituximab-containing chemotherapy. These findings will help optimize a preventive strategy, especially in hepatitis B virus endemic regions with limited healthcare resources. Clinical trial number: NCT 00931229.

Purification of infective bluetongue virus particles by immuno-affinity chromatography using anti-core antibody.

An immuno-affinity chromatography technique for purification of infective bluetongue virus (BTV) has been descried using anti-core antibodies. BTV anti-core antibodies (prepared in guinea pig) were mixed with cell culture-grown BTV-1 and then the mixture was added to the cyanogens bromide-activated protein-A Sepharose column. Protein A binds to the antibody which in turn binds to the antigen (i.e. BTV). After thorough washing, antigen-antibody and antibody-protein A couplings were dissociated with 4M MgCl2, pH6.5. Antibody molecules were removed by dialysis and virus particles were concentrated by spin column ultrafiltration. Dialyzed and concentrated material was tested positive for BTV antigen by a sandwich ELISA and the infectivity of the chromatography-purified virus was demonstrated in cell culture. This method was applied for selective capture of BTV from a mixture of other viruses. As group-specific antibodies (against BTV core) were used to capture the virus, it is expected that virus of all BTV serotypes could be purified by this method. This method will be helpful for selective capture and enrichment of BTV from concurrently infected blood or tissue samples for efficient isolation in cell culture. Further, this method can be used for small scale purification of BTV avoiding ultracentrifugation.

Immunization of cows with novel core glycolipid vaccine induces anti-endotoxin antibodies in bovine colostrum.

BACKGROUND: Translocation of gut-derived Gram-negative bacterial (GNB) lipopolysaccharide (LPS, or endotoxin) is a source of systemic inflammation that exacerbates HIV, cardiovascular and gastrointestinal diseases and malnutrition. The oral administration of bovine colostrum (BC) reduces endotoxemia in patients with impaired gut barrier function. Consequently, BC enriched in antibodies to LPS may ameliorate endotoxemia-related morbidities. We developed a detoxified J5 LPS/group B meningococcal outer membrane protein (J5dLPS/OMP) vaccine that induces antibodies against a highly conserved core region of LPS and protects against heterologous GNB infection. We now examine the ability of this vaccine to elicit anti-core endotoxin antibodies in BC. METHODS: Two cohorts of pregnant cows were immunized with this vaccine in combination with FICA (Cohort 1) or Emulsigen-D (Cohort 2) adjuvants. Antibody responses to the J5 core LPS antigen were measured in both serum and colostrum and compared to antibody levels elicited by a commercially available veterinary vaccine (J5 Bacterin) comprised of heat-killed Escherichia coli O111, J5 mutant bacteria, from which the J5 LPS was purified. RESULTS: The J5dLPS/OMP vaccine induced high titers of serum IgG antibody to J5 LPS in all seven cows. Both IgG and to a lesser extent IgA anti-J5 LPS antibodies were generated in the colostrum. The J5dLPS/OMP vaccine was significantly more immunogenic in mice than was the J5 Bacterin. BC enriched in anti-J5 LPS antibody reduced circulating endotoxin levels in neutropenic rats, a model of "leaky gut". CONCLUSION: The J5dLPS/OMP vaccine elicits high titers of serum anti-endotoxin antibodies in cows that is passed to the colostrum. This BC enriched in anti-core LPS antibodies has the potential to reduce endotoxemia and ameliorate endotoxin-related systemic inflammation in patients with impaired gut barrier function. Since this vaccine is significantly more immunogenic than the J5 Bacterin vaccine, this J5dLPS/OMP vaccine might prove to be more useful for veterinary indications as well.

[Strategies for preventing de novo hepatitis B infection after liver transplantation (II)]. / Estrategias para evitar la hepatitis B de novo postrasplante hepático (II).

Although active immunization against the hepatitis B virus (HBV) through vaccination constitutes a fundamental strategy in the prevention of infection by this virus, it is not effective in isolation for preventing de novo HBV infections in recipients of liver grafts from core antigen antibody (anti-HBc) positive donors. In this situation, the risk of developing de novo hepatitis B depends on the recipient's serological status. It has been shown that, for vaccinated patients and in the absence of prophylaxis with nucleoside/nucleotide analogues and/or hyperimmune gamma globulin, the prevalence and cumulative incidence of HBV infection after transplantation is an intermediate risk. The absence of a surface antigen antibody (anti-HBs) titer cutoff considered protective, the gradual reduction of these titers after vaccination, the presence of false positives for anti-HBs in patients undergoing infusion of blood products and escape mutations of the hepatitis B surface antigen (HBsAg) could explain this lack of efficacy. For this reason, it is recommended that vaccination protocols be implemented universally, along with the follow-up of the level of protection in patients with cirrhosis, adding prophylaxis with analogues when receiving a graft from an anti-HBc-positive donor. Clinical and serological surveillance alone can be considered for patients with anti-HBs levels greater than 200 mUI/mL after vaccination.

Prevalencia del virus de hepatitis B en donantes de sangre / Hepatitis B virus prevalence in blood donors

El Virus de la Hepatis B, representa un riesgo potencial en los receptores de sangre, por tal motivo, se pretende describir y analizar la prevalencia del VHB en los donantes de sangre. Como método, se utilizó la revisión bibliográfica, y se analizaron artículos de la biblioteca científica SciELO. En los resultados se evidenció una prevalencia de 1.12% (45) donantes positivos para el VHB en Irapuato México. En el estado Zulia en 46.563 donantes, 1.439 positivos (3.09%) para el antígeno frente al Core (anti-HBc) y 97 casos (0.208%) con antígeno de superficie (AgsHB) positivo. En el estado Sucre en 356 donantes, 41 (11.52%) positivos para el Anti-HBc, y 9 (2.53%) para HBsAg. Conclusión, los resultados indican que la prevalencia del VHB en los donantes de sangre continua siendo un factor de riesgo en los receptores, por lo cual es fundamental la historia clínica y la entrevista así como, la implementación de nuevas tecnologías en los bancos de sangre para minimizar el riesgo atribuible a la transfusión de sangre(AU)

The Hepatitis B Virus represents a potential risk to blood recipients in spite of the measures usually implemented to ensure blood safety. Therefore, we intend to describe and analyze the prevalence of HBV in blood donors. As a method we reviewed existing literature and analyzed scientific articles in the SciELO Library. The results showed a prevalence of 1.12% (45) HBV positive donors in Irapuato, Mexico. In the state of Zulia, Venezuela, out of 46,563 donors, 1,439 were positive (3.09%) for the B core antibody (anti-HBc) and 97 (0.208%) for the surface antigen (HBsAg). In Sucre state, out of 356 donors, 41 (11.52%) were positive foranti-HBc, and 9 (2.53%) for HBsAg. In conclusion, the results indicate that the prevalence of HBV in blood donors remains a risk factor for receptors, thus making imperative to emphasize the importance of the patient history and interview, as well as to implement the use new technologies in blood banks to minimize the risk(AU)

Diagnóstico serológico de la Hepatitis B / Serologic diagnostic of Hepatitis type B

Existen varios marcadores serolόgicos del virus de hepatitis B, siendo los más importantes el HBsAg, HBcAg, HBeAg y sus anticuerpos como el anti-HBs, el anticore, anticore IG M, anticore total y el anti e. Basado en la presencia del antígeno de superficie, se ha medido la seroprevalencia del virus de hepatitis B. Se considera a un país de alta prevalencia, si en la población estudiada los niveles son mayores del 8%. Se considera intermedia cuando oscila entre 2 y 8% y baja si es menos del 2%. La Caja Costarricense del Seguro Social cuenta con una red de 98 laboratorios, de los cuales 17 están dotados con los equipos necesarios para la determinación de los antígenos y anticuerpos del virus de la hepatitis B. Un estudio realizado en el 2005 determinó, en el ámbito nacional, una seroprevalencia del antígeno de superficie del 0.1% considerándose, a Costa Rica, como un país de baja incidencia del virus B. Se ha determinado también que San Isidro del General tiene una seroprevalencia intermedia del HBsAg. El Colegio Americano de Patólogos realiza un tipo de control externo para la CCSS en lo que se refiere a estas técnicas de laboratorio para la determinación y control del virus de hepatitis B. La aparición de los diferentes antigenos y anticuerpos mencionados se relacionan con momentos clínicos que se explican, considerándose de importancia, la persistencia del antígeno de superficie, por más de 6 meses, como un portador crónico.

The hepatitis B surface antigen (HBsAg), the hepatitis B core antigen (HBcAg), the hepatitis B e antigen (HBeAg) and its antibodies such as the anti-HBs, the anticore (HBcAb), the IG M anticore (HBcAb Ig M), the total anticore; and the anti HBe (HBeAb) are found to be the most important among several serological markers for hepatitis B virus. The seroprevalence of hepatitis B virus was measured based on the presence of surface antigen. Any country is considered highly prevalent if the levels of the population under study are higher than 8%. The prevalence is considered intermediate if levels are between 2 and 8%; and it is low if they are less than 2%. The Caja Costarricense de Seguro Social relies on a network of 98 laboratories each with the necessary equipment to find out the antigens and antibodies of hepatitis B virus. A study carried out in 2005 determined a surface antigen seroprevalence of 0.1% in all the country. Therefore, Costa Rica has been ranked as a country with low incidence of HBV. It has also been determined that San Isidro del General holds an intermediate seroprevalence of HBsAg. The American Pathologists College executes an external control for CCSS regarding the laboratory techniques used to determine and control the hepatitis B virus. The appearance of the different antigens and antibodies mentioned is related to clinical moments that are explained. It is also considered as important, the persistence of surface antigen for more than 6 months as a chronic carrier.