Identification of type A and B isolates of Epstein-Barr virus by polymerase chain reaction

A method is described for the identification of type A and type B isolates of Epstein-Barr virus (EBV) by means of the polymerase chain reaction. The use of three pairs of primers specific for genomic sequences coding for the two forms of EBV nuclear antigen (EBNA), 2A and 2B, and for a DNA sequence from the BamZ/BamR region allows the reliable and rapid detection of type A and B viruses in as little as 1000 EBV positive cells

Graphischer Ausdruck und Erkennen von Gefühlsqualitäten

Kritzeln ist ein spontaner graphischer Ausdruck. Seine Bedeutung in der Psychologie liegt vor allem in der Echtheit dieses Ausdrucks, der sich unverfälscht, direkt und eindeutig darstellt. Ob sich Gefühlserleben im Kritzelausdruck verdichtet und für einen Betrachter auch erfassbar ist, wird in einer zweigeteilten empirischen Untersuchung überprüft. Im ersten Teil drücken 21 Vpn drei Grundgefühle (Wut, Trauer, Freude) in Kritzeldarstellungen aus, die im zweiten Teil von weiteren 50 Vpn in drei Durchgängen mit steigendem Schwierigkeitsgrad überzufällig oft richtig identifiziert wurden. Dieses Ergebnis bestätigt ältere Untersuchungen und ist für die Allgemeine Psychologie im Rahmen der Ausdruckspsychologie bedeutsam, jedoch auch für die Klinische Psychologie auf diagnostischer und therapeutischer Ebene.Scribbling is a spontaneous graphic expression. It is significant for psychology due to its genuineness which represents the inner experience of man in an undistorted, direct and unequivocal manner. An ex-periment in two parts investigates whether emotional experiences are condensed in such scribbles and whether observers can understand them. In the first part 21 subjects express three basic emotions (anger, sadness, joy) in scribbles, which in the second part are identified in three trials of increasing difficulty by another 50 subjects. The number of correct identifications was higher than what would be expected by chance. This result confirms former studies and is important not only for general psychology but also for clinical psychology on a diagnostic and therapeutic level

Determinants of Long-term Protection After Hepatitis B Vaccination in Infancy: A Meta-analysis

Background: The duration of protection after hepatitis B vaccination in early infancy is unclear and may be related to vaccination schedule, dosage, vaccine type and population characteristics. Factors potentially influencing waning immunity were assessed. Methods: A systematic review was performed. The main outcomes were prevalence of anti-hepatits B antibodies >= 10 mIU/mL after primary or booster vaccination. Factors potentially influencing protection were assessed in an adjusted random-effects meta-analysis model by age for both outcomes. Results of both meta-analyses were combined in a prognostic model. Results: Forty-six studies reporting on the anti-hepatits B antibodies >= 10 mIU/mL 5 to 20 years after primary immunization and 29 on booster response were identified. The adjusted meta-analyses identified maternal carrier status (odds ratio OR]: 2.37 1.11; 5.08]), lower vaccine dosage than presently recommended (OR: 0.14 0.06; 0.30]) and gap time between last and preceding dose of the primary vaccine series (OR: 0.44 0.22; 0.86]) as determinants for persistence of anti-hepatits B antibodies >= 10. A lower vaccine dosage was also associated with failure to respond to booster (OR: 0.20 0.10; 0.38]). The prognostic model predicted long-term protection of 90% 77%; 100%] at the age of 17 years for offspring of noncarrier mothers vaccinated with a presently recommended dose and vaccination schedule. Conclusions: Based on meta-analyses, predictors of waning immunity after hepatitis B vaccination in infancy could be identified. A prognostic model for long-term protection after hepatitis B vaccination in infancy was developed

Urea-Mediated Cross-Presentation of Soluble Epstein-Barr Virus BZLF1 Protein

Soluble extracellular proteins usually do not enter the endogenous human leukocyte antigen (HLA) I–dependent presentation pathway of antigen-presenting cells, strictly impeding their applicability for the re-stimulation of protein-specific CD8+ cytotoxic T lymphocytes (CTL). Here we present for the Epstein-Barr virus (EBV) BZLF1 a novel strategy that facilitates protein translocation into antigen-presenting cells by its solubilisation in high molar urea and subsequent pulsing of cells in presence of low molar urea. Stimulation of PBMC from HLA-matched EBV-seropositive individuals with urea-treated BZLF1 but not untreated BZLF1 induces an efficient reactivation of BZLF1-specific CTL. Urea-treated BZLF1 (uBZLF1) enters antigen-presenting cells in a temperature-dependent manner by clathrin-mediated endocytosis and is processed by the proteasome into peptides that are bound to nascent HLA I molecules. Dendritic cells and monocytes but also B cells can cross-present uBZLF1 in vitro. The strategy described here has potential for use in the development of improved technologies for the monitoring of protein-specific CTL

Comparative purification and characterization of hepatitis B virus-like particles produced by recombinant vaccinia viruses in human hepatoma cells and human primary hepatocytes

This study describes the comparative expression and purification of hepatitis B surface antigen (HBsAg) particles produced upon infection of human primary hepatocytes and human hepatoma cell lines (HuH-7 and HepG2) with recombinant vaccinia viruses. The highest levels of HBsAg expression were found in HuH-7 hepatoma cells following infection with recombinant vaccinia viruses, which contain the S gene under control of a 7.5 k-promoter. Four different methods for purification of the HBsAg particles were examined: isopycnic ultracentrifugation, sucrose cushion sedimentation, isocratic column gel filtration, and binding to anti-HBs-coated microparticles. The highest degree of purity of HBsAg particles was reached by the method based on anti-HBs-coated microparticles. The resulting product was > 98% pure. Biochemical analysis and characterization of purified HBsAg particles were performed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), western blotting, and electron microscopy. The HBsAg, purified from human hepatoma cell lines and from human primary hepatocytes, consisted of both the non-glycosylated (p25) and the glycosylated (gp27) form and assembled into typical 22-nm particles, and thus may be of great interest and importance for research, diagnostics, and medical treatments

Test Performance Characteristics of Anti-HEV IgG Assays Strongly Influence Hepatitis E Seroprevalence Estimates

Hepatitis E virus (HEV) seroprevalences of 0.3%–53% were reported from industrialized countries. Because these estimates may be influenced by detection assays, this study compares 3 frequently used tests for HEV detection: the MP Diagnostics HEV immunoglobulin G (IgG) enzyme-linked immunosorbent assay (ELISA), the Axiom Diagnostics HEV IgG enzyme immunoassay (EIA), and the Mikrogen recomLine HEV IgG assay. Sera from 200 healthy healthcare workers and 30 individuals with acute HEV infection were analyzed. Among the healthy individuals, HEV IgG was found in 4.5% by the MP Diagnostics assay, in 29.5% by the Axiom Diagnostics assay, and in 18% by the Mikrogen assay. Among individuals with acute HEV infection, positive results were obtained for 83.3%, 100%, and 96.7%, respectively. Thus, the 3 assays show clear differences in diagnostic sensitivity

BCG Induced Necrosis of the Entire Bladder Urothelium

AbstractInstillation therapy with attenuated tuberculosis bacteria (BCG) can significantly reduce rates of recurrence of non-muscle invasive bladder cancer. Local and systemic side effects such as dysuria, irritative voiding symptoms or partial bladder contracture and systemic inflammation were reported. A 75 year-old male patient with recurrent non muscle invasive bladder cancer developed necrosis of the entire bladder urothelium more than six years after BCG instillation immunotherapy. The resulting irritative voiding symptoms and low bladder capacity required radical cystectomy. BCG instillation can cause severe side effects, which develop gradually and eventually need radical surgical therapy such as cystectomy without tumor recurrence

Isolation of Subtype 3c, 3e and 3f-Like Hepatitis E Virus Strains Stably Replicating to High Viral Loads in an Optimized Cell Culture System

The hepatitis E virus (HEV) is transmitted via the faecal–oral route in developing countries (genotypes 1 and 2) or through contaminated food and blood products worldwide (genotypes 3 and 4). In Europe, HEV subtypes 3c, 3e and 3f are predominant. HEV is the leading cause of acute hepatitis globally and immunocompromised patients are particularly at risk. Because of a lack of cell culture systems efficiently propagating wild-type viruses, research on HEV is mostly based on cell culture-adapted isolates carrying uncommon insertions in the hypervariable region (HVR). While optimizing the cell culture system using the cell culture-adapted HEV strain 47832c, we isolated three wild-type strains derived from clinical specimens representing the predominant spectrum of HEV in Europe. The novel isolates 14-16753 (3c), 14-22707 (3e) and 15-22016 (3f-like) replicate to high viral loads of 108, 109 and 106.5 HEV RNA copies/mL at 14 days post-inoculation, respectively. In addition, they could be kept as persistently infected cell cultures with constant high viral loads (~109 copies/mL) for more than a year. In contrast to the latest isolates 47832c, LBPR-0379 and Kernow-C1, the new isolates do not carry genome insertions in the HVR. Optimization of HEV cell culture identified amphotericin B, distinct salts and fetal calf serum (FCS) as important medium supplements. Overconfluent cell layers increased infectivity and virus production. PLC/PRF/5, HuH-7-Lunet BLR, A549 and HepG2/C3A supported replication with different efficiencies. The novel strains and optimized cell culture system may be useful for studies on the HEV life cycle, inactivation, specific drug and vaccine development

Characteristics of immune memory 10–15 years after primary hepatitis B vaccination

Background and aims: The definition of immune memory after hepatitis B vaccination is still under debate. Therefore, we analysed hepatitis B surface antigen (HBsAg)-specific memory in more detail by investigating the kinetics of humoral and cellular responses after hepatitis B booster vaccination. Methods: The anti-HBs kinetics of 23 individuals with anti-HBs titres below 10 IU/I, who had been vaccinated 10-15 years ago, was monitored at day 0, 3, 7,14 and 28 after booster vaccination. HBsAg-specific IFN gamma- and IL5-secreting cells in enriched CD4(+) fraction were measured at day 0, 7 and 28 post-booster by enzyme-linked immunospot assay (ELISpot). Results: 22 of 23 subjects showed similar anti-HBs kinetic curves, including 3 of 4 subjects who did not reach anti-HBs titres of 10 IU/I. The steep anti-HBs increase started between day 3 and 7 and peaked around day 14. A plateau or only minimal changes were visible between day 14 and 28. 17.4% of subjects showed pre-booster cellular responses, and this rate had increased to 47.8% and 56.5% after 7 and 28 days, respectively. The kinetic patterns of T cell responses differed considerably among subjects. A dominance of Th2 responses (IL5 secretion) over Th1 responses (IFN gamma secretion) could be observed. Conclusions: The presence of B cell memory could be shown by a typical anamnestic anti-HBs response curve after a booster dose in all but one individual. In contrast, T cell responses to booster vaccination, which occurred in approximately 50% of participants, were rather heterogeneous. (C) 2015 Elsevier Ltd. All rights reserved