Electronic Library of Scientific Literature - © Academic Electronic Press

Acta virologica

Volume 48 / 2004 / number 3

Here you can download FULL version of Acta Virologica 02/2004 in PDF format (parts are ordered by name of author): 
Part 1 (Angalova.pdf – 175 kB),  Part 2 (Dhinakar_Raj.pdf – 126 kB),  Part 3 (Hamdollah_Zadeh.pdf – 313 kB), Part 4 (Kundu.pdf – 1,318 MB), Part 5 (Kuno.pdf – 100 kB), Part 6 (Perez-Gracia.pdf – 60 kB), Part 7 (Rudenko.pdf – 578 kB), Part 8 (Ruszczyk.pdf – 139 kB), Part 9 (Sanyal.pdf – 173 kB), Part 10 (Sidorkiewicz.pdf – 395 kB), Part 11 (Subr.pdf – 189 kB).
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Review

Articles

Short Comunications

Erratum

A SURVEY OF THE RELATIONSHIPS AMONG THE VIRUSES NOT CONSIDERED ARBOVIRUSES, VERTEBRATES, AND ARTHROPODS

G. KUNO

Arbovirus Diseases Branch, Division of Vector-Borne Infectious Diseases, National Center for Infectious Diseases, Centers for Disease Control and Prevention, P.O. Box 2087, Fort Collins, CO 80522-2087, USA

Received June 18, 2004; accepted September 13, 2004

Summary. – No single group of organisms demonstrates more extensive and diverse associations with animal viruses than the phylum Arthropoda. Compared with the well-recognized relationship found in arboviruses, however, most of the atypical arthropod-vertebrate relationships of the viruses normally not considered arboviruses have received much less attention, as they remain in the marginal areas of interest for most researchers in animal virology, veterinary medicine, medical entomology, and invertebrate pathology. However, this comprehensive review of the information gathered from several branches of virology by profession reveals highly valuable information potentially useful in the fields of research ranging from investigations of the mode of transmission of poorly understood or emerging viral diseases to studies of the evolution of biological transmission of animal viruses by arthropod vectors. The observations and data obtained for the animal virus relationships with arthropods and vertebrates outside the boundaries of arboviruses, in turn, can be used to re-examine more closely the definition of arboviruses. With increasing number of reports challenging one of the basic tenets of the definition of arbovirus (requirement of viremia in vertebrate host) and others describing virus-host relationships that complicate the definition of arbovirus, the accumulated information clearly demonstrates the difficulty of defining the boundaries of arboviruses.
Key words: animal virus; arthropod; vertebrate; arbovirus; virus-host relationship

Acta virologica 48: 135 – 143, 2004

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TRANSGENIC RESISTANCE TO TOBACCO RINGSPOT VIRUS

A. HAMDOLLAH ZADEH1, G.D. FOSTER2

1 Plant Pest and Disease Institute, Agricultural Research and Education Organization, Teheran, Iran;
2 School of Biological Sciences, University of Bristol, Woodland Road, Bristol BS8 1UG, UK

Received October 7, 2003; accepted July 30, 2004

Summary. – The coat protein (CP) gene including the 3'-untranslated region (UTR) of RNA2 of a cherry isolate of Tobacco ringspot virus (TRSV) was utilized in a CP-mediated resistance (CP-MR) strategy. To facilitate construction of plant expression vectors the sequence context of the CP gene translation initiation codon was modified at the 5'-end of the coding sequence by including an initiation codon. The gene was ligated to a version of the Cauliflower mosaic virus (CaMV) 35S promoter with a duplicated enhancer. The cloned CP gene was used to transform Nicotiana tabacum cv. Xanthi, as a systemic and local lesion host. The transgenic plants showed different level of resistance ranging from complete resistance to reduction in symptom severity post inoculation with the cherry isolate of TRSV. A CP gene transcript was detected in different tissue of transgenic lines, but translation product was undetectable by Western blot analysis or enzyme-linked immunosorbent assay (ELISA). Interestingly, 100% of seed transmission was blocked in a resistant line, which offers important prospects for engineering TRSV into economically important crops as soybean with 100% seed transmission.
Key words: coat protein gene; coat protein-mediated resistance; Tobacco ringspot virus; post transcriptional gene silencing

Acta virologica 48: 145 – 152, 2004

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THE EFFECT OF INTERLEUKIN-6 ON HEPATITIS B VIRUS REPLICATION IN PERIPHERAL BLOOD MONONUCLEAR CELLS IN VITRO

M. SIDORKIEWICZ1, B. JÓŹWIAK1, Z. SULOWSKA2, J. GREGER1, U. LEWANDOWSKA1

1 Department of Medical Biochemistry, Medical University of Lodz, 92-215 Lodz, Poland; 2 Microbiology and Virology Center, Polish Academy of Science, Lodz, Poland

Summary. – Although the major target organ for hepatitis B virus (HBV) is the liver, the possibility of infection of peripheral blood mononuclear cells (PBMCs) with HBV has also been reported. This study was performed to analyze the course of HBV infection of PBMCs and to investigate the influence of interleukin-6 (IL-6) on the efficiency of infection of PBMCs with HBV in vitro. PBMCs isolated from a healthy donor were infected by exposing to a HBsAg-, HBeAg-positive serum in the presence or absence of exogenous IL-6. The efficiency of infection was estimated by HBV DNA determination in the cells and medium in the course of infection. The results of this study show that the presence of IL-6 during the PBMCs infection with HBV increased the efficiency of this infection.
Key words: Hepatitis B virus; peripheral blood mononuclear cells; interleukin-6; PCR

Acta virologica 48: 153 – 158, 2004

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COMPLETE NUCLEOTIDE SEQUENCE ANALYSIS OF A VACCINE STRAIN AND A FIELD ISOLATE OF FOOT-AND-MOUTH DISEASE VIRUS SEROTYPE ASIA1 WITH AN INSERTION IN VP1 GENOMIC REGION

A. SANYAL, J.K. MOHAPATRA, R. MANOJ KUMAR, S. BISWAS, D. HEMADRI, C. TOSH, G.P. SABARINATH, S.K. GUPTA, M. MITTAL, P. GIRIDHARAN, S.K. BANDYOPADHYAY

Project Directorate on Foot-and-Mouth Disease, Indian Veterinary Research Institute Campus, Mukteswar-Kumaon, Nainital 263 138, Uttaranchal, India

Received March 10, 2004; accepted August 17, 2004

Summary. – Complete nucleotide sequences except the poly (C) tract and poly (A) tail of a vaccine strain (IND 491/97) and an atypical field isolate (IND 321/01) of Foot-and-mouth disease virus (FMDV) serotype Asia1 are described. Amino acid (aa) sequence analysis of the VP1 protein of the field isolate revealed that the latter has 212 instead of 210 or 211 aa found in the so far available sequences of other FMDV isolates of Asia1 serotype. The insertion was localized in the hypervariable region of aa 130–160 of VP1 protein. Nucleotide sequencing of the entire genome was therefore carried out to detect changes in other parts of the genome, if any, besides VP1, which could contribute to its fitness. An 8.16 kb sequence of IND 491/97 and an 8.162 kb sequence of IND 321/01 were compared with each other and also with the known sequence of IND 63/72, another vaccine strain of serotype Asia1. Comparison of the entire polyprotein coding (L to 3D) region of IND 321/01 with those of the two Asia1 vaccine strains (IND 63/72 and IND 491/97) revealed no significant differences. A similar comparison of IND 491/97 with IND 63/72 revealed variability across the entire length of the genome. In addition to the capsid-coding region, sequence variability was also observed in non-structural proteins albeit to different extent. This study shows that in the gene pool of serotype Asia1 at least three groups of isolates/strains are present with respect to the length of VP1 protein.
Key words: field isolate IND 321/01; Foot-and-mouth disease virus; genome; serotype Asia1; vaccine strain IND 491/97; nucleotide sequence

Acta virologica 48: 159 – 166, 2004

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TICK-BORNE ENCEPHALITIS VIRUS-SPECIFIC RT-PCR – A RAPID TEST FOR DETECTION OF THE PATHOGEN WITHOUT VIRAL RNA PURIFICATION

N. RUDENKO1,2, M. GOLOVCHENKO1,2, V. CIHLÁŘOVA2, L. GRUBHOFFER1,2

1Faculty of Biological Sciences, University of South Bohemia and 2Institute of Parasitology, Academy of Sciences of the Czech Republic, Branišovská 31, 370 05 České Budějovice, Czech Republic

Received December 12, 2003; accepted July 30, 2004

Summary. – Among diseases transmitted by ticks in the Czech Republic, tick-borne encephalitis (TBE) caused by Tick-borne encephalitis virus (TBEV) and Lyme disease caused by Borrelia burgdorferi spirochete are most important. We propose an effective and specific test for detection of TBEV in a single tick or a pool of ticks based on the detection of TBEV RNA using an RT-PCR technique without RNA purification. The method is very sensitive with the detection limit of about 14 fg TBEV RNA in total RNA obtained from brain suspension from suckling mice infected with TBEV per reaction. The primers were derived from the 5'-terminal non-coding region, a highly conserved part of the virus. The method was successfully applied to field-collected ticks in detecting TBEV RNA. This method can be used in studies of several aspects of TBEV: epidemiology, screening of natural foci, circulation and detection of virus genome sequences in clinical materials.
Key words: Ixodes ricinus; Tick-borne encephalitis virus; Chelex® 100 resin; RT-PCR; SYBR Green

Acta virologica 48: 167 – 171, 2004

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A SIMPLIFIED RT-PCR-BASED DETECTION OF RECOMBINANT PLUM POX VIRUS ISOLATES

Z. ŠUBR, S. PITTNEROVÁ, M. GLASA

Institute of Virology, Slovak Academy of Sciences, Dúbravská cesta 9, 845 05 Bratislava, Slovak Republic

Received May 25, 2004; accepted July 30, 2004

Summary. – Closely related natural Plum pox virus (PPV) isolates derived from homologous RNA recombination between PPV-D and PPV-M have been recently identified and shown naturally spread in several European countries. As their serological properties were identical with those of conventional PPV-M isolates, they could be detected only by combined analysis of at least two different genome parts. To simplify the detection of such recombinants primers specific to PPV-M and PPV-D sequences with binding sites located on both sides of the recombination crossover situated in the C-terminal part of NIb were designed. They were used for direct differentiation of PPV-M, PPV-D and their recombinants by reverse transcription–polymerase chain reaction (RT-PCR). This method is convenient for identification of a recombinant PPV in single as well as mixed infection with PPV-M or PPV-D.
Key words: Sharka; recombination; molecular diagnostic

Acta virologica 48: 173 – 176, 2004

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DETECTION OF BEET YELLOWS VIRUS BY RT-PCR AND IMMUNOCAPTURE RT-PCR IN TETRAGONIA EXPANSA AND BETA VULGARIS

K. Kundu, P. Ryšánek

Department of Plant Protection, Czech University of Agriculture, 165 21 Prague 6, Czech Republic

Received May 28, 2004; accepted July 30, 2004

Summary. – Two sensitive methods, RT-PCR with phenol-extracted RNA or Triton X-100-released RNA and immunocapture RT-PCR (IR-RT-PCR) were used for the detection of Beet yellows virus (BYV) in young and old leaves of Tetragonia expansa and sugar beet (Beta vulgaris) and in sugar beet roots. Four oligonucleotide primer pairs proved suitable for the detection of BYV. The release of BYV RNA with Triton X-100 was shown to be a very effective and easy as compared to isolation of total RNA by phenol extraction with the same or higher sensitivity of subsequent PCR. Using the Triton X-100 release of RNA and IC-RT-PCR the sensitivity of detection was so high that pg amounts of BYV RNA occurring in dilutions up to 10-6 of saps from young Tetragonia and sugar beet leaves could be detected.
Key words: Beet yellows virus; immunocapture-RT-PCR; RT-PCR; Triton X-100

Acta virologica 48: 177 – 182, 2004

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COMPARISON OF HEMAGGLUTINATION INHIBITION TEST AND ELISA IN QUANTIFICATION OF ANTIBODIES TO EGG DROP SYNDROME VIRUS

G. DHINAKAR RAJ, S. RATNAPRABA, K. MATHESWARAN, K. NACHIMUTHU

Department of Animal Biotechnology, Madras Veterinary College, Tamil Nadu Veterinary and Animal Sciences University,
Chennai – 600 007, India

Received October 17, 2003; accepted August 5, 2004

Summary. – A single-serum dilution ELISA for egg drop syndrome (EDS) virus-specific antibodies was developed. In testing 425 chicken sera it was found to have a 93.6% sensitivity and 98.7% specificity relative to a hemagglutination inhibition (HI) test. The correlation coefficient for ELISA and HI titers was 0.793. The ELISA was efficacious in quantification of both vaccinal and infection antibodies and could routinely be used for screening large numbers of field sera.
Key words: egg drop syndrome virus; antibodies; quantification; ELISA; HI test; correlation coefficient

Acta virologica 48: 183 – 187, 2004

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EQUINE HERPES VIRUS 2 INFECTION IN HORSE POPULATIONS IN POLAND

A. RUSZCZYK, A. CYWINSKA, M.W. BANBURA

Department of Preclinical Sciences, Faculty of Veterinary Medicine, Warsaw Agricultural University, Ciszewskiego 8, 02-786 Warsaw, Poland

Received February 2, 2004; accepted August 17, 2004

Summary. – The prevalence of Equine herpesvirus 2 (EHV-2) infections in the horse populations in Poland was investigated. Peripheral blood leukocytes (PBLs) of 139 horses were tested. The animals were divided into four groups: clinically healthy horses, horses suffering from respiratory disorders, mares with a recent abortion and horses with diagnosed ataxia. Thirty-four virus isolates were obtained from leukocytes of the tested animals by cocultivation with equine dermal cells and were identified as EHV-2 by PCR using primers for the gB gene of EHV-2 and/or primers for the sequence located upstream of the gene homologous to the equine interleukin 10 (IL-10) gene. These results indicate that EHV-2 is prevalent in horse populations in Poland. As the virus was most frequently isolated from horses with respiratory disorders its etiological importance may be considered.
Key words: Equine herpesvirus 2; horse populations; interleukin 10; prevalence; cocultivation; PCR

Acta virologica 48: 189 – 192, 2004

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EXPRESSION OF CELLULAR PROTEINS BCL-XL , XIAP AND BAX INVOLVED IN APOPTOSIS IN CELLS INFECTED WITH HERPES SIMPLEX VIRUS 1 AND EFFECT OF PAVINE ALKALOID (-)-THALIMONINE ON VIRUS-INDUCED SUPPRESSION OF APOPTOSIS

A. ANGELOVA1, T. TENEV2, T. VARADINOVA1

1 Laboratory of Virology, Faculty of Biology, University of Sofia, 8 Dragan Tzankov Blvd., 1421 Sofia, Bulgaria;
2 Institute of Cancer Research, Chester Beatty Laboratories, London SW3 6JB, UK

Received March 29, 2003; accepted September 13, 2004

Summary. – The expression of three cellular proteins involved in the modulation of apoptosis, namely antiapoptotic Bcl-XL and XIAP and proapoptotic Bax, was investigated in cells infected with Herpes simplex virus 1 (HSV-1). To assess whether the regulation of apoptosis in virus-infected cells depends on strain specificity the wild-type (wt) strain Victoria and the mutant R-100 resistant to acyclovir (ACV) were used. In addition, the expression of Bcl-XL, XIAP and Bax was studied in cells infected with HSV-1 and treated with pavine alkaloid (-)-thalimonine. Our previous work has demonstrated that (-)-thalimonine irreversibly inhibits the replication of wt HSV-1 in cultured cells. Our data showed that (-)-thalimonine down-regulates the expression of viral proteins UL17, VP11-12, VP22, VP24 and g134.5, and affects negatively the posttranslational processing of glycoproteins D (gD) and G (gG). As both g134.5 and glycoprotein D possess antiapoptotic activity, we investigated whether the antiviral effect of the alkaloid could also be due to its ability to suppress the antiapoptotic activity of the virus. Our results demonstrated that: (i) the virus induced overexpression of antiapoptotic proteins Bcl-XL and XIAP; (ii) (-)-thalimonine reduced their overexpression, and (iii) this effect was stronger with the acyclovir resistant mutant R-100 than with the wt virus. Taken together, these data suggest that: (i) the virus abolishes apoptosis by means of virus-induced up-regulation of cell-specific prosurvival proteins Bcl-XL and XIAP, and (ii) (-)-thalimonine, apart from affecting essential viral targets, inhibits the infectious progeny production via restoration of apoptosis during viral replication.
Key words: Herpes simplex virus 1; apoptosis; Bax; Bcl-XL; XIAP; (-)-thalimonine; biological response modifiers

Acta virologica 48: 193 – 196, 2004

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DETECTION OF HEPATITIS E VIRUS IN PATIENTS SERA IN SOUTHERN SPAIN

M.T. Pérez-Gracia1, M.S. García-Valdivia2, F. Galán2, M.A. Rodríguez-Iglesias2

1 Departamento de Atención Sanitaria, Salud Pública y Sanidad Animal, Facultad de Ciencias Experimentales y de la Salud. Universidad Cardenal Herrera-CEU, 46113 Moncada, Valencia, Spain;
2 Laboratorio Microbiología, Hospital Universitario de Puerto Real, Universidad de Cádiz, Cádiz, Spain

Received June 7, 2004; accepted September 13, 2004

Summary. – The aim of the present study was to detect acute Hepatitis E virus (HEV) infection in patients with abnormal alanine transaminase (ALT) in which other viral hepatitis infections had been excluded in southern Spain, an area adjacent to regions where this disease is endemic. Of 336 sera tested 30 (8.92%) were positive for IgM antibodies against HEV (anti-HEV IgM) and 7 (2.08%) were negative in a repeated assay. Immunoblot analysis (IBA) was applied to the 37 positive sera in the first assay; its results were positivity for 26 (7.73%), ambiguous for 5 and negative for 6 sera. Amplification of ORF1 and ORF2 of HEV by means of nested RT-PCR was carried out with the 37 sera that were either positive or ambiguous by ELISA; a positive result was obtained only with one serum for the ORF2 protein. IgM antibodies against the HEV ORF2 protein could be a useful marker in the diagnosis of acute infection and a substitute for the determination of viral RNA in serum; this is of both diagnostic and epidemiological importance as it would allow the patients transmitting the infection to be recognized by means of a simple determination of antibodies. The sequence of the ORF2 fragment of HEV occurring in samples taken from both humans and animals amplified in this study has considerable homology with the sequences of HEV strains/isolates of European origin. These results demonstrate that an autochthonous HEV circulates in Spain.
Key words: Hepatitis E virus; IgM antibodies against HEV; ELISA; immunoblot analysis; nested RT-PCR

Acta virologica 48: 197 – 200, 2004

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