Electronic Library of Scientific Literature Electronic Library of Scientific Literature
Volume 42 / December 1998 / number 6
R. DURMAZ, A. AYDIN, M. KÖROGLU, B. DURMAZ, H. ÇIRALIK
Department of Clinical Microbiology and
Department of Pathology, Inönü University, Faculty of Medicine, Malatya 44069,Turkey
Summary. – To investigate whether Epstein-Barr virus (EBV) is associated with ordinary gastric carcinoma and to determine its genotype, samples from ordinary gastric carcinoma from 65 patients (40 males, 20 females) and 21 endoscopic biopsies from 7 individuals with non-neoplastic mucosa were analysed using one-stage and nested (two-stage) polymerase chain reaction (PCR) assays. The nested PCR assay yielded 56.9% (37/65) and 52.3% (11/21) positivity for the ordinary gastric carcinoma and control cases, respectively; these results were significantly than those of the one-stage PCR assay (13.5% and 0%, respectively). The EBV positivity showed similar rate in male and female patients (60% versus 52% , P >0.05). The dominant genotype of EBV was 1 (92%) followed by mixture of 1+2 (5.4%) and 2 (2.6%). In conclusion, similar positivity rates of EBV in neoplastic and non-neoplastic tissues suggest that the relationship of this virus to the ordinary gastric carcinoma is not clear.
Key words: Epstein-Barr virus; polymerase chain reaction; gastric adenocarcinoma;
virus genotype
Acta virologica 42: 359 – 363, 1998
E. KOVÁČOVÁ, J. KAZÁR, D. ŠPANĚLOVÁ
Institute of Virology, Slovak Academy of Sciences, Dúbravská cesta 9, 842 46
Bratislava, Slovak Republic;
Institute of Preventive and Clinical Medicine, Bratislava;
State Veterinary Institute, Bratislava
Summary. – Comparison of four serological tests (complement fixation (CF) test, microagglutination (MA) test, microimmunofluorescence (MIF) test, and enzyme-linked immunosorbent assay (ELISA)) for detection of post-infection antibody response in human and animal sera revealed a low sensitivity of the CF test with acute Q fever human, goat and sheep sera but not with chronic Q fever human sera and sera of aborting cows. The remaining three tests gave similar results with human (both acute and chronic) and cow sera, but the ELISA was more sensitive than the MA and MIF tests with goat and sheep sera. A treatment of phase I Coxiella burnetii (C.b.) cells with chloroform-methanol, potassium periodate and trichloroacetic acid (TCA), and mild acidic hydrolysis did not result in increase of the sensitivity of the tests when compared with the natural phase I and phase II C.b. cells, respectively. The suitability of various C.b. antigen preparations for the abovementioned serological tests with various sera is discussed.
Key words: Coxiella burnetii; Q fever; antigenic preparations; complement fixation;
microagglutination; microimmunofluorescence; enzyme-linked immunosorbent assay
Acta virologica 42: 365 – 368, 1998
A. GIBADULINOVÁ, V. ZELNÍK, L. REISEROVÁ, E. ZÁVODSKÁ, M. ZAŤOVIČOVÁ, F.
ČIAMPOR,
S. PASTOREKOVÁ, J. PASTOREK
Institute of Virology, Slovak Academy of Sciences, Dúbravská cesta 9, 842 46 Bratislava, Slovak Republic
Summary. – We have cloned and characterised a cDNA encoding Z protein of recently identified MX strain of lymphocytic choriomeningitis virus (LCMV) persistently infecting human MaTu cells. Deduced amino acid sequence of LCMV MX Z protein showed 88.9% identity with that of the LCMV Armstrong (ARM) strain and 80.9% identity with that of the LCMV Traub (TRA) strain. It contained conserved zinc-binding RING finger domain and C-terminal proline-rich region. Northern blot analysis of total RNA from MaTu cells revealed presence of abundant truncated forms of L RNA. Z protein-specific rabbit antibodies were produced to glutathione S-transferase (GST)-Z fusion protein expressed in E. coli and used for the detection of Z protein in MaTu cells. Western blot and immunofluorescence analyses detected relatively high levels of Z protein indicating its role in maintenance of persistent LCMV.
Key words: lymphocytic choriomeningitis virus; Z protein; MaTu cells
Acta virologica 42: 369 – 374, 1998
R. DURMAZ, A. AYDIN, M. KÖROGLU, H. AKER, I.H. ÖZERCAN, E. ATIK, S. ARICI
Department of Clinical Microbiology and
Department of Pathology, Inönü University, Faculty of Medicine, 44069 Malatya, Turkey;
Department of Pathology, Cumhuriyet University, Faculty of Medicine, Sivas,Turkey;
Department of Pathology, Firat University, Faculty of Medicine, Elazig, Turkey
Summary. – In order to determine the positivity rate and genotype of Epstein-Barr virus (EBV) in cases with Hodgkin’s disease (HD) in Turkey, 40 tissue specimens from HD patients were analysed. Ten non-lymphoid tissue samples from individuals without any evidence for lymphoma were used as controls. The cases with HD included 33 males and 7 females with a mean age of 28 years. Nodular sclerosis was the most prevalent histological subtype (16/40) followed by mixed cellularity (10/40), lymphocyte predominance (9/40), and lymphocyte depletion (5/40). After histopathological evaluation, deparafinisation and lysis of the specimens, one-stage polymerase chain reaction (PCR) and two-stage (nested) PCR assays were performed with the primers common for both EBV genotypes and the primers specific for EBV types 1 and 2, respectively. EBV DNA was detected in 22 of 40 (55%) cases with HD and in 1 of 10 (10%) control specimens. The distribution of EBV DNA positivity according to the histological subtypes was as follows: 10 of 16 (62.5%) for nodular sclerosis, 3 of 5 (60%) for lymphocyte depletion, 5 of 9 (55.6%) for lymphocyte predominance, and 4 of 10 (40%) for mixed cellularity. Although most of the HD patients were males of 15 – 34 years of age, there were no significant differences between EBV positivities obtained from different sex and age groups. The rates of EBV genotypes were 82% for type 1, 9% for type 2, and 9% for both types, respectively.
Key words: Epstein-Barr virus; virus genotype; Hodgkin’s disease; polymerase
chain reaction
Acta virologica 42: 375 – 381, 1998
H.S. PEREIRA, M.A. REBELLO
Carlos Chagas Filho Institute of Biophysics, and
Institute of Microbiology, Department of Virology, Federal University of Rio de Janeiro,
21941-590 Rio de Janeiro, RJ, Brazil
Summary. – The antibiotic cerulenin, an inhibitor of lipid synthesis, was shown to suppress Mayaro virus replication in Aedes albopictus cells at non-cytotoxic doses. Cerulenin blocked the incorporation of [3H]glycerol into lipids when present at any time post infection (p.i.). Cerulenin added at the beginning of infection inhibited the synthesis of virus proteins. However, when this antibiotic was added at later stages of infection, it had only a mild effect on the virus protein synthesis. The possibility that cerulenin acts by blocking an initial step in the Mayaro virus replication after virus entry and before late viral translation is discussed.
Key words: Mayaro virus; Aedes albopictus cells; lipid synthesis; cerulenin
Acta virologica 42: 383 – 388, 1998
R.G. DAMLE, L.R. YEOLEKAR, B.L. RAO
National Institute of Virology, 20-A, Dr. Ambedkar Road, Post Box No.11, Pune 411 001, Maharashtra, India
Summary. – Monoclonal antibodies (MAbs) against an Indian strain (804994) and an Egyptian strain (E 101) of West Nile virus (WNV) were prepared in mice. Nine MAbs against the 804994 strain and 5 MAbs against E 101 strain were obtained. All 14 MAbs reacted with the envelope (E) protein of WNV in an immunoblot assay. They were tested by an enzyme-linked immunosorbent assay (ELISA) for their cross-reactivity with WNV, Japanese encephalitis virus (JEV) and Dengue-2 virus (DEN-2), and for their reactivity in haemagglutination-inhibition (HAI) test. Based on these results MAbs were broadly grouped into three groups, namely WNV-specific HAI-positive, WNV-JEV cross-reactive HAI-positive, and WNV-JEV cross-reactive HAI-negative MAbs. The antigenic cross-reactivity between twelve WNV strains isolated from different geographical regions and their respective hosts was assessed using these MAbs in HAI and complement fixation (CF) tests. The strain analysis by CF distinguished Indian from South African strains. However, a similarity between some Indian and South African strains in HAI was observed. E 101 strain appeared to have antigenic similarity with Indian as well as South African strains. Overall it appears that antigenically similar strains of WNV are prevalent in India. A single heterogenous domain was apparent on the epitope map of WNV deduced by ELISA additivity test.
Key words: West Nile virus; monoclonal antibody; strain variation; epitope mapping
Acta virologica 42: 389 – 395, 1998
P.K. GUPTA, M. SAINI, S.K. BANDYOPADHYAY, S.K. GARG
National Biotechnology Centre and
Division of Biochemistry and Food Science, Indian Veterinary Research Institute, Izatnagar
243 122, UP, India;
Department of Veterinary Microbiology, G.B. Pant University of Agriculture and Technology,
Pantnagar, UP, India
Summary. – Glycoprotein C (gC) of bovine herpesvirus type 1 (BHV-1) is a major viral glycoprotein expressed at high level on the surface of infected cells and on the virion envelope. This glycoprotein is also a major target of immune response at both humoral and cellular levels. The plasmid pRSV-gC having complete coding gene for BHV-1 gC was transfected into MDBK cells and the expression of gC in these cells was detected by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate (SDS-PAGE) and immunoblot analysis. Transcription of the gC gene was confirmed by reverse transcription-polymerase chain reaction (RT-PCR) of total RNA isolated from transfected cells. MDBK cells expressing BHV-1 gC were used as antigen in enzyme-linked immunosorbent assay (ELISA) of antibodies against BHV-1 in field sera. The results were found comparable (92.44%) with those obtained with BHV-1 purified antigen.
Key words: bovine herpesvirus type 1; glycoprotein C; expression; ELISA; RT-PCR
Acta virologica 42: 397 – 400, 1998
A. CHIBA, T. SUZUTANI, M. SAIJO, S. KOYANO, M. AZUMA
Department of Microbiology and
Department of Pediatrics, Asahikawa Medical College, 4-5 Nishikagura, Asahikawa 078-8510,
Japan
Summary. – To analyze the difference in the degree of divergence between genes from identical herpesvirus species, we examined the nucleotide sequence of genes from the herpes simplex virus type 1 (HSV-1) strains VR-3 and 17 encoding thymidine kinase (TK), deoxyribonuclease (DNase), protein kinase (PK; UL13) and virion-associated host shutoff (vhs) protein (UL41). The frequency of nucleotide substitutions per 1 kb in TK gene was 2.5 to 4.3 times higher than those in the other three genes. To prove that the polymorphism of HSV-1 TK gene is common characteristic of herpesvirus TK genes, we compared the diversity of TK genes among eight HSV-1, six herpes simplex virus type 2 (HSV-2) and seven varicella-zoster virus (VZV) strains. The average frequency of nucleotide substitutions per 1 kb in the TK gene of HSV-1 strains was 4-fold higher than that in the TK gene of HSV-2 strains. The VZV TK gene was highly conserved and only two nucleotide changes were evident in VZV strains. However, the rate of nonsynonymous substitutions in total nucleotide substitutions was similar among the TK genes of the three viruses. This result indicated that the mutational rates differed, but there were no significant differences in selective pressure. We conclude that HSV-1 TK gene is highly diverged and analysis of variations in the gene is a useful approach for understanding the molecular evolution of HSV-1 in a short period.
Key words: herpes simplex virus; varicella-zoster virus; nucleotide sequence;
polymorphism; molecular evolution
Acta virologica 42: 401 – 407, 1998
S.A. BOPEGAMAGE, A. PETROVIČOVÁ
Institute of Preventive and Clinical Medicine, Limbová 14, 833 01 Bratislava, Slovak Republic
Summary. – Cytokines have been suggested to play an important role in the pathogenesis of type I diabetes mellitus through their direct and indirect effects on the pancreatic islet cells. We studied the time course of tumour necrosis factor alpha (TNF-alpha) and glucose levels in the sera of mice infected with coxsackie B4 and A7 viruses. Two correlating peaks of TNF-alpha and glucose were found. These results suggest the involvement of TNF-alpha in the damage of the insulin producing cells and thus an immunity-related inflammatory process.
Key words: coxsackie viruses; TNF-alpha; glycaemia; mice; diabetes mellitus
Acta virologica 42: 409 – 412, 1998
K. SUBI
Department of Virology, Institute of Experimental and Clinical Medicine, Hiiu 42, 11626 Tallinn, Estonia
Summary. – During the period of 1981–1997, a total of 13,110 patients with acute respiratory viral infections were tested simultaneously for influenza virus A and B, parainfluenza, adenovirus and respiratory syncytial virus (RSV) infections. Of 3,800 laboratory-confirmed cases mixed infections were established in average in 27%. The incidence varied in dependence on the season between 12% and 36%. All infections were involved in mixed infections, yet their participation greatly varied in different seasons. During a season when the incidence of two respiratory viral infections increased, the case number of their mixed infections also increased. We assume that respiratory viruses widely co-circulating among human population also simultaneously infected the same persons. Therefore a proportion of respiratory viral diseases may represent in fact mixed infections which, in turn, may lead to the emergence of mixtures of different viruses and their spread in humans.
Key words: respiratory viral infections; laboratory diagnosis; mixed infections
Acta virologica 42: 413 – 415, 1998
P. EBBESEN, V. ZACHAR
Department of Virus and Cancer, Danish Cancer Society, Gustav Wieds Vej 10, 8000 Aarhus
C, Denmark;
Institute of Virology, Slovak Academy of Sciences, Bratislava, Slovak Republic
Summary. – An evidence is accumulating that the oxygen tension exerts significant effect on the virus replication in vitro. When the in vitro oxygen tension is maintained at an in vivo physiological level, as a rule higher yields of human viruses are seen than at conventional culturing with access of an unphysiologically high oxygen concentration in ambient air. Although not fully understood, possible explanation for this phenomenon may be provided by a lowered interferon (IFN) output and increased cell replication which is often optimal at physiological oxygen tension. Furthermore, an indirect evidence suggests that the expression of some virus receptors is affected by oxygen tension. Also, the antiviral cell-mediated immunity is likely to be found oxygen tension-dependent as both the NK and cytotoxic T cell activities towards uninfected target cells are oxygen tension-sensitive. At present, the in vitro work with viruses at physiological oxygen tensions is hampered by the fact that cells adapt in the course of several weeks to the new oxygen tension. Whether viruses may adapt to different oxygen tensions is not clear. Workbenches combining safety in manipulation with hazardous viruses and the convenience of controlled gas atmosphere during both manipulation and long-term incubation have been developed. It is suggested that the in vitro virology should ensure that the physiological oxygen tension is better mimicked in the in vivo processes. Much work is to be done to determine the molecular interactions between oxygen tension-sensitive elements of the cell and infecting viruses. Of no lesser importance are the questions regarding the role of oxygen in virus tissue tropism, the cost-benefit of virus production at different oxygen tension levels, and the potential significance of oxygen tension for delivering gene effects to the selected target tissues.
Key words: oxygen tension; virus replication; in vitro conditions; equipment
Acta virologica 42: 417 – 421, 1998