Immunofluorescence test for sensitive detection of varicella-zoster virus-specific IgG: an alternative to fluorescent antibody to membrane antigen test.

A highly sensitive indirect fluorescence antibody test (IFAT) has been developed on the basis of varicella-zoster virus (VZV)-infected human lung carcinoma (A549) cells and evaluated for the determination of immunity to VZV. Different serum panels with negative, low, moderate or high anti-VZV IgG levels detected by the fluorescent antibody to membrane antigen (FAMA) assay were investigated. As a result, the sensitivity and the specificity of IFAT were 100% compared to FAMA test. In anti-VZV IgG-positive sera, a significant correlation between the results of FAMA procedure and IFAT could be shown. However, there were considerably higher antibody titers by the IFAT than by FAMA. Whereas the FAMA test had a detection limit of 250 mIU/ml anti-VZV IgG, the limit of detection of IFAT was 50 mIU/ml. In conclusion, the IFAT using VZV-infected A549 cells as antigen allows a highly sensitive, specific, and rapid detection of anti-VZV IgG class antibodies. This simple technique can replace the labor-intensive FAMA procedure for laboratory determination of immunity to VZV.

Seroprevalence of varicella-zoster virus in the German population.

The present study was conducted to generate data on the epidemiology of varicella-zoster virus (VZV) infections in Germany as a basis for health economic evaluations of varicella vaccination strategies. The survey was designed as a cross-sectional, age-stratified study of the VZV seroprevalence in the German population. The status of immunity of 4602 individuals a aged 0 to >70 years was investigated by means of an indirect enzyme immunoassay and the fluorescent antibody to membrane assay. After waning of maternal antibodies over the period of 6-9 months seropositivity rates remained low by the end of the 1st year of life. By the age of 4-5 years 62.5% (95% CI; 56.0-68.5) of the pre-school children had already been infected with VZV and at the age of 10-11 years 94.2% (95% CI; 91.0-96.0) of children were positive for anti-VZV antibodies. Among the age-group of >40 years old, only few individuals were susceptible for VZV. The median antibody levels to VZV did not significantly decline with increasing age. In comparison with figures of previous studies the age-specific seroprevalence data presented here do not provide evidence for an upward shift in the age distribution of varicella in Germany. Since the majority of VZV infections occurs during the early childhood, the best option to reduce the circulation of wild-type VZV in the population would be the immunization of young children.

Clinical, radiologic, demographic, and occupational aspects of hand osteoarthritis in the elderly.

OBJECTIVE: Osteoarthritis (OA) of the hand is common in elderly patients. The aim of this study was to characterize OA frequency, severity, and distribution and to trace interrelationships between these findings and the demographic, occupational, and medical data from elderly Jewish nonrheumatologic patients. METHODS: Study participants were 253 consecutive patients admitted to a geriatric center for a variety of nonrheumatic medical conditions. Excluded patients were those with rheumatoid arthritis; neurologic, orthopedic, or other conditions that would interfere with symmetric hand function; and mental or medical states that would interfere with history taking and radiographic studies. Patient occupations were graded as workload degree (on a scale of 1 to 3) and as the total occupational score (workload degree multiplied by the duration of each job). Clinical findings of Heberden nodes, Bouchard nodes, and malignment, graded on a scale of 0 to 3, were summed as the clinical OA score. Hand radiographs were independently read (modified Altman method), grading 5 parameters in each joint on a scale of 0 to 3, summed as a radiologic OA score. Statistical analyses included the Student t test, chi(2) test, ANOVA, Pearson correlation, and partial correlation coefficients. RESULTS: Among 253 elderly patients (171 women, 82 men; mean age, 79 years) OA was frequent (occurring in about 80% of patients), involving most severely the second and third distal interphalangeal, right first interphalangeal, and both first carpometacarpal joints. The prevalence of OA was similar in women and men, with higher scores in women, and reached significance only in the distal interphalangeal joints. Metacarpophalangeal joints were more involved in men. Age had a clear influence on OA scores. Ethnicity affected OA severity, with Ashkenazi Jews having significantly higher scores than Sepharadi Jews. Dominant hands had significantly higher global OA scores as well as isolated joint scores (except for the first carpometacarpal joint). Occupational load, housekeeping tasks, and the number of children did not influence the total or specific joint OA scores. Associated conditions such as obesity, diabetes, hypothyroidism, and chondro calcinosis were not associated with more pronounced OA. CONCLUSIONS: Hand OA was prevalent in our elderly cohort, and its severity was influenced by inherent traits such as age, female gender, ethnicity, and handedness. In contrast, acquired factors such as workload, number of children, and associated diseases did not appear to influence OA expression.

Serological diagnosis of Epstein-Barr virus infection by novel ELISAs based on recombinant capsid antigens p23 and p18.

A new pair of Epstein-Barr virus ELISAs (Biotest Anti-EBV VCA IgG and VCA IgM ELISA) was evaluated for usefulness for routine diagnosis of acute EBV infections. The ELISAs are based on two viral capsid antigens (VCA), p23 (BLRF2, full-length) and p18 (BFRF3, carboxy-half), that are combined by autologous gene fusion. In total, 179 sera were tested in direct comparison with classical VCA immunofluorescence assays (IFA). With the help of clinical data and additional reference serology, i.e., heterophile antibodies, anti-EA IgG (IFA) and anti-EBNA-1 IgG (ELISA), the patients were divided into the following categories: seronegatives (46), acute primary infections (67), previous infections (39), suspected reactivations (20) and constellations with intermediate serological patterns (7). The VCA IgG and VCA IgM ELISAs showed overall agreement to IFA of 95.0% and 94.4%, respectively. The calculated analytical performance (sensitivity; specificity) of VCA IgG and VCA IgM was 94.0%; 97.8% and 97.1%; 96.5%, respectively. A certain delay in seroconversion of anti-p23-p18 IgG may account for a significant difference in sensitivity of the VCA IgG ELISA between primary (88.4%) and previous infections (100%). In summary, the new recombinant VCA ELISAs yielded good correlation to VCA IFA and in combination with EBNA-1 IgG allow rapid, sensitive, and specific diagnosis of infectious mononucleosis or EBV immune status in general.

Seroprevalence of herpes simplex virus type 1 and type 2 in selected German populations-relevance for the incidence of genital herpes.

This study was carried out to determine the prevalence of antibodies to herpes simplex virus types 1 (HSV-1) and 2 (HSV-2) in selected German populations, such as blood donors, hospital patients, and human immunodeficiency virus (HIV)-seropositive individuals. Serum samples collected between 1996 and 1998 were tested by enzyme immunoassays using monoclonal antibody-selected native gG1 and gG2 as antigens and an immunoblot using type-specific recombinant glycoproteins. Equivocal results were resolved by an "in-house" Western blot assay. The prevalence of HSV-1 antibodies increased steadily with age and reached high levels of >/=88% among subjects 40 years of age or older. In the sample of patients and blood donors, the HSV-2 seroprevalence was 12.8% (95% CI = 11.9-13.8%). About 81% of the HSV-2 seropositive subjects were coinfected with HSV-1. When adjusted for age, there was no difference in the HSV-2 seroprevalence between hospital patients and blood donors. The HSV-2 seroprevalence was significantly higher among women (15%) than among men (10.5%), yielding a female : male odds ratio of 1.5 for hospital patients and of 1.67 for blood donors. Among the HIV-infected population, 91.1% were seropositive for HSV-1 and 47.9% for HSV-2. HIV-infected women have a significantly higher risk of HSV-2 infection than men (odds ratio [OR] = 3.22; 95% confidence ratio [CI] 1.99-5.20). In conclusion, although the rate of infections with HSV-2 is relatively low in the German population, attention should be given to the further development in adolescents, especially in view of a possible decrease of HSV-1 seroprevalence in childhood.

Antiviral efficacies of famciclovir, valaciclovir, and brivudin in disseminated herpes simplex virus type 1 infection in mice.

The animal model of necrotic hepatitis caused by HSV-1 infection in juvenile mice was used to compare the efficacies of the oral antiherpes agents famciclovir (FCV), valaciclovir (VACV) and brivudin (BVDU). The experimental infection allows the measurement of viral replication in the liver by macroscopic lesions and the evaluation of mortality from encephalitis. Mice intravenously inoculated with a highly virulent clinical HSV-1 isolate were orally treated by gavage over a period of 3 days starting on day 2 post infection. The reference drug acyclovir (ACV) was administered subcutaneously. Necrotic hepatitis was significantly (p < 0.01) reduced by treatment with FCV, VACV and ACV at a dose of 50 mg/kg per day divided into 3 doses. No significant effect was achieved with BVDU at 200 mg/kg per day. Treatment with FCV at 50 mg/kg per day, ACV at 100 mg/kg per day, and VACV at 200 mg/kg per day significantly (p < 0.001) decreased mortality in mice. BVDU treatment at 200 mg/kg per day did not reduce mortality but significantly prolonged (p < 0.05) the survival time.

Oral brivudin vs. intravenous acyclovir in the treatment of herpes zoster in immunocompromised patients: a randomized double-blind trial.

The efficacy of oral brivudin vs. intravenous acyclovir was compared in a randomized multicentered study under double-blind conditions using the double-dummy technique. Forty-eight patients with a herpes zoster rash less than 72 hours in duration were entered in the study. Brivudin was given as one 125-mg tablet every 6 hours. Acyclovir was infused over 1 hour at a dose of 10 mg/kg every 8 hours. Treatment was continued for 5 days. There was no significant difference between the treatment groups when analyzed in terms of new lesion formation, increase in the area of rash within the primary dermatome, cutaneous dissemination, and affection of mucous membranes or visceral organs. Both treatment regimes were also equally effective in the time to full crusting of lesions. Oral brivudin and intravenous acyclovir were well tolerated by most patients. There was no need to interrupt the treatment in any case. As effective as intravenous acyclovir in the treatment of herpes zoster, oral brivudin offers the potential for outpatient treatment of herpes zoster in immunocompromised patients.

Malignant lymphomas induced by an Epstein-Barr virus-related herpesvirus from Macaca arctoides--a rabbit model.

Animal models for Epstein-Barr virus (EBV) are restricted to some species of new-world monkeys which develop malignant lymphoid tumours or benign lymphoproliferative diseases after virus inoculation. Similar pathological features were induced in rabbits by the EBV-related herpesvirus of Macaca arctoides (HVMA). In this study 17 of 32 rabbits infected with varying amounts of HVMA produced from MAL-1 cells developed lymphoproliferative disorders. In 13 rabbits high-grade malignant lymphomas were detected, 4 rabbits revealed the histopathological feature of lymphoid hyperplasia. These lymphoproliferations were shown to be associated with HVMA by PCR and by the expression of EBV-like RNAs (EBER) in 14 and 10 cases, respectively. The homology in the polymerase gene region between DNA from EBV and HVMA, and from HVMA and the malignant tissue was found to be 94.8% and 100%, respectively. All the infected animals produced antibodies to antigens corresponding to early and late EBV proteins. By studying the HVMA expression in MAL-1 cells EBV-like proteins expressed in latency (EBNA1 and EBNA2) and in the lytic cycle (VCA, EA) were detected. Our findings suggested that HVMA caused a symptomatic infection in rabbits with pathological features that fit the conditions of an animal model suitable for testing antiviral drugs and vaccines against EBV.

Serological diagnosis of infectious mononucleosis using three anti-Epstein-Barr virus recombinant ELISAs.

A new Epstein-Barr virus (EBV) ELISA system (Biotest Anti-EBV recombinant) was evaluated for usefulness for routine diagnosis of EBV primary infection. The assay system is composed of three different microtest plates coated with three highly purified recombinant EBV antigens. The early antigens p138 (BALF2, truncated) and p54 (BMRF1, whole sequence) are used as a mixture for testing IgM (assay 1) and IgG (assay 2) antibodies. In addition, the EBNA-1 antigen p72 (BKRF1, carboxy-half) is used for detecting IgG antibodies (assay 3). Three panels of sera were examined in direct comparison with standard immunofluorescence (IF): Specimens of (i) 120 infectious mononucleosis (IM) patients, (ii) 60 patients with acute CMV infection, toxoplasmosis or rheumatic disease, respectively, and (iii) 185 healthy blood donors as a control group. 119 IM patients were clearly recognized as having acute primary infection (sensitivity 99.2% compared to VCA-IgM by IF). Three apparently false-positive results were obtained with patients of other diseases and none within the control group (specificity 98.8%). The data suggest that the recombinant ELISA can be used advantageously for standardized rapid diagnosis of acute EBV primary infection.

[Infections after bone marrow transplantation in childhood]. / Infektionen nach Knochenmarktransplantationen im Kindesalter.

In 66 children having undergone bone marrow transplantation (BMT) the occurrence of infections was studied retrospectively. Bacterial infections were mostly found in the early period after transplantation before marrow engraftment. The analysis of positive blood cultures showed a dominance of gram-positive bacteria, especially of coagulase-negative staphylococci. Cytomegalovirus (CMV) infections were most important, because of its high rate and the risk of CMV associated interstitial pneumonia (IP), two patients suffered from. Infections from herpes simplex virus (HSV), varizella zoster virus (VZV) and Epstein Barr virus (EBV) had no influence on prognosis. In fungal infections the systemic aspergillosis was the most important complication. To increase the effectiveness and safety of therapy the serum levels of antibiotics and antifungal drugs should be determined.

[Clinical aspects of acute cytomegalovirus infection in adults without immune deficiency]. / Zur Klinik der akuten Cytomegalievirusinfektion beim nichtimmungeschwächten Erwachsenen.

The clinical pictures of 11 adults with cytomegalovirus-infection were analyzed. Characteristic symptom was fever of unknown origin lasting up to 50 days. Indications of an accompanying hepatitis were found in all patients and confirmed by increased serum levels of the transaminases which for al short time exceeded 5 mumols/s/l (AST) and 8 mumols/s/l (ALT) only in 2 patients. Mononucleosislike pictures, however, have not been seen. One patient developed neurological symptoms. The diagnosis was made by demonstrating IgM and IgG antibodies by way of the fluorescence antibody test. An antiviral therapy has not been introduced.

[Detection of Epstein-Barr virus in the tonsils in infectious mononucleosis]. / Nachweis des Epstein-Barr-Virus in Tonsillen bei infektiöser Mononukleose.

Tonsils of 50 patients with infectious mononucleosis were examined for the presence of Epstein-Barr virus nuclear antigens (EBNA) and in 11 cases for the presence of Epstein-Barr virus (EBV) nucleic acid sequences. In tonsillar tissue of 42 patients less than 1 to 40 per cent EBNA-positive cells could be demonstrated by anticomplement immunofluorescence. 10 out of 11 tonsils examined by in situ hybridization contained less than 1 to 50 per cent cells with EBV nucleic acid sequences. The histological examination indicated that cells labelled by in situ hybridisation are in the majority proliferating B-lymphocytes and to a small extent cells of tonsillar epithelium.

Antibodies to Epstein-Barr virus-induced early antigens in blood donors.

IgG class antibodies to the early antigen complex (EA; D + R components) of the Epstein-Barr virus (EBV) were found by indirect immunofluorescence in titres greater than or equal to 1:10 in 196 (49.4%) out of 397 blood donors and titres greater than or equal to 1:20 in 84 (21.2%) of these subjects. Anti-EA titres of greater than or equal to 1:2560 were detected in 6 sera, anti-EA(D) only in 17 donors (4.3%). Additional EBV-specific antibody tests were performed in sera with anti-EA-IgG titres of greater than or equal to 1:20 (n = 84), including IgM class antibodies to virus capsid antigen (anti-VCA-IgM). Specific IgM revealed active EBV infection in 12 blood donors. There was no correlation between the presence of anti-VCA-IgM and the magnitude of anti-EA-IgG titres. Therefore, it seems to be impossible to define the threshold titre for EA antibodies indicating active EBV infection. For this purpose probably a titre increase should be demonstrated in paired sera.

CMV infections in bone marrow transplanted patients--evaluation of prophylaxis with Cytotect (CMV hyperimmuneglobulin).

Cytomegalovirus (CMV) infection is a frequent and clinically important infection following bone marrow transplantation. Candidates for this study were patients admitted for transplantation: 22 patients received bone marrow from a HLA-identical, MCR-nonreactive sibling, in 9 patients an autologous BMT was performed. The anti-CMV IgG (Cytotect) was administered at a dosage of 1 ml/kg on days -7, 13, 33, 53, 73 and 93 after BMT. 5 patients in the very beginning of our BMT program did not receive Cytotect. Patients were given random blood products from the bloodbank not tested for CMV positivity. Active CMV infection or seroconversion in our patients was defined as a rise in IgG titer against the late antigen of fourfold or more or an IgM increase. In the allogeneic BMT group the pretransplant serological status was in 6 cases negative in recipients and donor, in 7 patients positive in recipients and negative in donors, and in 4 patients positive in recipients and donors. Of the 6 patients seronegative in recipients and donors, 3 developed active infection and of the 7 patients pretransplant positive with seronegative donors 3 developed active infection and 4 latent infections during the period from 2 to 100 days following grafting. 1 patient out of the group transplanted in third partial remission of AML developed interstitial pneumonia and died on day +30.4 of the 4 cases with seropositivity of recipients and donors developed active CMV infection. Of 9 patients with autologous transplantation 6 patients were pretransplant seropositive. 3 of these 6 developed active infection and 2 latent infection 30 to 180 days after grafting.(ABSTRACT TRUNCATED AT 250 WORDS)

Oxygen tension within the arterial wall: relationship to altered bioenergetic metabolism and lipid accumulation.

Oxygen tension (pO2) was measured in upper thoracic arteries and in muscular foci at the celiac bifurcation from atherosclerosis-susceptible White Carneau and atherosclerosis-resistant Show Racer pigeons at 6, 12, and 24 weeks of age. At each site the pO2 was measured within adventitial (outer), medial (middle), and subendothelial (inner) zones with polarographic oxygen microelectrodes while the animals were under dissociative anesthesia. The atherosclerosis-resistant Show Racer pigeons exhibited no significant age or site differences in pO2 levels. The medial pO2 was approximately 25 mm Hg, while the adventitial and subendothelial zones had pO2 levels of approximately 30 mm Hg. The pO2 profiles through the various zones of the arterial wall of 6-week-old atherosclerosis-susceptible White Carneau pigeons were similar to those of the Show Racers. However, significantly lower oxygen tension was found in the subendothelial zone from 12- and 24-week White Carneau celiac bifurcations (27 and 21 mm Hg) than in corresponding zones in Show Racer sites. Also, the medial zone in 24-week White Carneau celiac foci exhibited a significantly lower pO2 (20 mm Hg) than the medial zone of younger White Carneau. When correlated with previously described biochemical and morphological changes in White Carneau pigeons, the findings presented herein indicate that focal decreases in oxygen availability within arterial tissue occur after the onset of spontaneous atherogenesis, and probably result from decreased diffusion of oxygen into the arterial wall.

[Detection of Epstein-Barr virus nucleic acid sequences in non-Hodgkin's lymphoma]. / Nachweis von Epstein-Barr-Virus-Nukleinsäuresequenzen in Non-Hodgkin-Lymphomen.

Tumor tissue specimens of 25 patients with various entities of Non-Hodgkin's lymphomas were examined for the presence of Epstein-Barr virus (EBV) nucleic acids and EBV nuclear antigens (EBNA) by in situ hybridization and anticomplement immunofluorescence. In tumor cells of five patients EBV nucleic acid and EBNA were demonstrated. The histopathologic examinations revealed in these cases one Burkitt's lymphoma, two centroblastic lymphomas and each one lymphoplasmocytic and lymphocytic lymphoma.

Effect of (E)-5-(2-bromovinyl)- and 5-vinyl-1-beta-D-arabinofuranosyluracil on Epstein-Barr virus antigen expression in P3HR-1 cells: comparison with acyclovir.

The effect of (E)-5-(2-bromovinyl)- and 5-vinyl-1-beta-D-arabinofuranosyluracil (BrVaraU, VaraU) in comparison to 9-(2-hydroxyethoxymethyl)guanine (ACV) on the proliferation of human lymphoblastoid P3HR-1 cells in culture and on the expression of Epstein-Barr virus capsid antigen (VCA) in the same cells was evaluated. After 7 days of cell growth, at 100 mumol/l the total number of new generations in drug-treated cultures was similar or 5 and 10% below that in drug-free control cultures, for VaraU, ACV, and BrVaraU, respectively. During the same time the percentage of VCA-expressing cells decreased from 6.3% in drug-free cultures to 1.3, 1.5, and 2.0% in cultures treated with VaraU, ACV and BrVaraU, respectively. In VaraU-treated cultures a further decrease in the percentage of VCA-positive cells down to 0.5% was revealed 7 days after drug removal. VaraU was also effective in reducing the proportion of VCA-expressing cells at 10 and 1 mumol/l. At 14 days after drug removal, the inhibitory effect of ACV was nearly reversed, whereas BrVaraU showed a prolonged VCA- suppressing effect.

[Herpesvirus infections in children with acute lymphatic leukemia]. / Herpesvirusinfektionen bei Kindern mit akuter lymphatischer Leukämie.

33 children with acute lymphatic leukemia and 33 healthy controls were longitudinally studied for herpesvirus infections. Active herpes simplex-virus, varicella-zoster virus and cytomegalovirus (CMV) infections were more frequent in patients than in controls. CMV and Epstein-Barr virus infections were often inapparent or associated with infections of the upper respiratory tract. Analysis of serological datas revealed a coincidence of active CVM infection and lethal course of the leukemia. This may be a result of the immunodeficiency in leukemia patients caused by disease and therapy. An additional influence of the immunosuppressive effect of active CMV infections on the course of the disease is discussed.