Electronic Library of Scientific Literature Electronic Library of Scientific Literature
Volume 44 / April 2000 / number 3
M.N. JOSHUA, Y. QI, Y. FU-HUA, H. YONG-XIU
Institute of Virology, Wuhan University, Wuhan 430072, P.R. China
Summary. – Recombinant transposing plasmids pFH24 and pFH41 were constructed by cloning the human immunodeficiency virus 1 (HIV-1) p24 and gp41 genes, respectively, into the transposing vector pFastBacHTa. Recombinant bacmids rBH24 and rBH41 were obtained by transposing pPolh/p24 and pPolh/gp41 expression cassettes from recombinant plasmids pFH24 and pFH41, respectively. Recombinant viruses rAcH24 and rAcH41 were generated by transfection of the Spodoptera frugiperda (Sf9) cells with the DNAs of plasmids rBH24 and rBH41, respectively. Analysis of the expressed p24 or gp41 proteins with an antiserum to HIV-1 (HIV-1 antiserum) by an enzyme-linked immunosorbent assay (ELISA) and dot blot assay showed high biological activity of these proteins; p24 was more active than gp41. Also a Western blot analysis showed stronger bands for p24 than for gp41. The high reactivities of p24 and gp41 with the HIV-1 antiserum suggest that these proteins could also be used as specific standard antigens in HIV-1 diagnostics.
Key words: HIV-1; p24; gp41 genes; proteins; bacmid, baculovirus; Sf9 cells
Acta virologica 44: 125 – 130, 2000
A. SAMAD, S.K. RAJ, A. SRIVASTAVA, G. CHANDRA, P.V. AJAYAKUMAR, M. ZAIM, B.P. SINGH
Division of Microbiology and Plant Pathology, Central Institute of Medicinal and
Aromatic Plants, Lucknow 226 015, India;
Plant Virus Laboratory, National Botanical Research Institute, Lucknow, India
Summary. – A cucumber mosaic virus isolate was found to be associated with mottle crinkle and severe mosaic disease of Egyptian henbane (Hyoscyamus muticus L.). The virus has been characterized as an Indian isolate of cucumber mosaic virus (CMV) based on non-persistent transmission by aphid, presence of 28-nm isometric particles, capsid protein of 26 K and single-stranded tripartite RNA genome with a subgenomic RNA (RNA 4). There was no evidence of satellite RNA genome. The isolate showed a strong serological relationship with S and A strains of CMV (CMV-S and CMV-A) in double diffusion test. A band of the 26 K capsid protein was also detected by Western blot analysis using antibodies specific to CMV-S.
Key words: cucumber mosaic virus; Hyoscyamus muticus; mottle crinkle; severe
mosaic; Western blot analysis
Acta virologica 44: 131 – 136, 2000
F. BROCCOLO, R. IULIANO, A.M. CAREDDU, R. TROVATO, S. LICO, P.L. BLANC, F. MAZZOTTA, L. CECCHERINI-NELLI
Department of Experimental Pathology, Medical Biotechnology, Infectious Diseases and
Epidemiology, University of Pisa, 56127 Pisa, Italy;
AIDS Immunopathogenesis Unit, DIBIT, San Raffaele Scientific Institute, Milano, Italy;
Division of Infectious Diseases, S.M. Annunziata Hospital, Antella, Florence, Italy
Summary. – Cerebrospinal fluid (CSF) samples from 49 acquired immunodefficiency disease syndrome (AIDS) patients with a central nervous system (CNS) disease were examined by polymerase chain reaction (PCR) to evaluate the association between the positivity for cytomegalovirus (CMV) and Epstein-Barr virus (EBV), and clinical diagnosis of a CNS disease. Frequency and clinical relevance of detection of DNA of human herpesviruses 6 (HHV-6), 7 (HHV-7) and 8 (HHV-8) were also determined. DNA of one or more of the following viruses was found in 26 of 49 patients (53%): CMV in 16 (33%), EBV in 13 (27%), human herpesvirus 6 (HHV-6) in 2 (4%), human herpesvirus 7 (HHV-7) in 1 (2%), and human herpesvirus 8 (HHV-8) in 1 (2%). The CMV detection was significantly associated with encephalitis and peripheral neuropathy (7/16 vs. 2/33, p = 0.003), while EBV with primary CNS lymphoma (P-CNSL) (8/13 vs. 0/36, p <0.0001). HHV-6 DNA was found in CSF of two patients with neuroradiological features suggestive of cerebral lesions. HHV-8 or HHV-7 DNA was detected in the CSF of patients with unexplained neurological symptoms. This study confirms that the PCR analysis of CSF is a valid tool for the diagnosis of neurological diseases associated with CMV and EBV. On the other hand, HHV-6, HHV-7 and HHV-8, instead, were rarely detected in CSF of AIDS patients and have certainly no correlation with the CNS disease found.
Key words: AIDS; CMV; CNS disease; EBV; HHV-6; HHV-7; HHV-8;
Acta virologica 44: 137 – 143, 2000
C.W. CHOI, S.H. PARK, J.K. CHOI, K.H. RYU, W.M. PARK
Department of Biology and
BioMed RRC, Pai Chai University, Taejon 302-735, South Korea;
Division of Biological Environment, Kangwon National University, Chunchon, South Korea;
Department of Horticultural Science, Seoul Women’s University, Seoul, South Korea;
Graduate School of Biotechnology, Korea University, Seoul, South Korea
Summary. – In order to determine the detection limit for chemically treated virions by gel electrophoresis, reverse transcription–polymerase chain reaction (RT-PCR) and infectivity assay, tobacco mosaic virus (TMV) exposed to various concentrations of chemicals was studied. When virions were exposed to 0.2 N HCl for 30 mins, partially degraded TMV particles were observed by gel electrophoresis. Under the same exposure, a major RT-PCR amplified DNA product corresponding to the target size of 806 bp, which decreased as a function of time, could be detected for up to 60 mins of exposure. When virions were treated with NaOH (0.02 N or higher normality) for 5 mins, partially degraded virions were detected by gel electrophoresis, exhibiting multiple band patterns. Exposure of the virions to 0.1 N NaOH for 5 mins revealed severely degraded viral RNA, but disappearance of the amplified RT-PCR products was apparent during 30–60 mins of exposure. Therefore, these data showed clearly the difference in the detection limit of gel electrophoresis and that of RT-PCR for the degraded viral RNA. In addition, the infectivity assay showed that the number of local lesions in Nicotiana rustica were significantly reduced by more than 95% when the virus was exposed to 0.2 N HCl for 15 mins or 0.1 N NaOH for 10 mins. From these results we conclude that loss of infectivity was not related to that of PCR product. Other chemical disinfectants such as phenol or formalin were also found to be effective to reduce the virus infectivity, but a corresponding degradation of viral RNA was detected by neither gel electrophoresis nor RT-PCR.
Key words: tobacco mosaic virus; chemical degradation; UV irradiation;
infectivity; RT-PCR; gel electrophoresis
Acta virologica 44: 145 – 149, 2000
R.J. PHILLPOTTS, T. LESCOTT, A.J. GATES, L. JONES
DERA, Biological Sciences Department, Chemical and Biological Defense Sector, Porton
Down, Wiltshire, SP4 0JQ, U.K.;
NERC Institute of Virology, Mansfield Road, Oxford OX1 3SR, U.K.
Summary. – Although it is unlikely that large-scale vaccination against smallpox will ever be required again, it is conceivable that the need may arise to vaccinate against a human orthopoxvirus infection. A possible example could be the emergence of monkey poxvirus (MPV) as a significant human disease in Africa. Vaccinia virus (VV) recombinants, genetically modified to carry the immunogenic proteins of other pathogenic organisms, have potential use as vaccines against other diseases present in this region. The immune response to parental wild-type (wt) or recombinant VV was examined by binding and functional assays, relevant to protection: total IgG, IgG subclass profile, B5R gene product (gp42)-specific IgG, neutralizing antibodies and class 1-mediated cytotoxic lymphocyte activity. There was a substantial reduction in the immune response to VV after scarification with about 108 PFU of recombinant as compared to wt virus. These data suggest that to achieve the levels of immunity associated with protection against human orthopoxvirus infection, and to control a possible future outbreak of orthopoxvirus disease, the use of wt VV would be necessary.
Key words: vaccinia virus; immunity; TK–; scarification
Acta virologica 44: 151 – 156, 2000
D. LIU, Y. QI, X. SUN, Z. LIU, S. HE
Institute of Virology, Wuhan University, Wuhan 430072, P.R. China;
College of Life Sciences, Central China Normal University, Wuhan, P.R. China
Summary. – The nucleocapsid protein gene (vp39) of a Chinese isolate of Bombyx mori nucleopolyhedrovirus (BmNPV-Ch), namely an open reading frame (ORF) of 1050 bp that codes for a polypeptide of 39 K (VP39) consisting of 350 amino acids was sequenced. The homology of the nucleotide (nt) and amino acid sequences of vp39 and VP39, respectively, of BmNPV-Ch and a Japanese isolate of BmNPV (BmNPV-Ja) were found to be 97.5% and 97.1%, respectively. The BmNPV-Ch vp39 is nine nucleotides longer than that of BmNPV-Ja vp39 due to insertion of CGA at nt 625 and GTCGGC at nt 985–910. There are differences in 17 nucleotides causing a few substitutions of amino acids which slightly modify the secondary structure of BmNPV-Ch. It indicates that the main part of the secondary structure of VP39 is a folded structure containing high proportion of beta-sheet and beta-turn units. A dot blot hybridization analysis revealed the existence of a homologous transcript of BmNPV-Ch vp39 in Sf9 cells infected with Autographa californica multiple nucleocapsid nucleopolyhedrovirus (AcMNPV).
Key words: nucleocapsid protein; VP39; gene; BmNPV; Chinese isolate; Japanese
isolate; secondary structure
Acta virologica 44: 157 – 161, 2000
M. BYSTRICKÁ, L. GAŠPAROVIČOVÁ, D. STANEKOVÁ, M. MOKRÁŠ, L. SOLÁRIKOVÁ, G. RUSS
Institute of Virology, Slovak Academy of Sciences, Dúbravská cesta 9, 842 45
Bratislava, Slovak Republic;
National Reference Centre for Prevention of HIV/AIDS, Institute of Preventive and Clinical
Medicine, Bratislava, Slovak Republic;
Department of AIDS, Clinic of Infectious Diseases, Bratislava, Slovak Republic
Summary. – We determined the prevalence of antibodies to herpes simplex virus 2 (HSV-2, HSV-2 antibodies) in sera of homosexual men either positive for human immunodeficiency virus 1 (HIV-1, HIV+, a group of 27 sera) or negative for HIV-1 and HIV-2 (HIV–, a group of 52 sera) in Slovakia. Antibodies to HSV-2 glycoprotein G-2 (gG-2, gG-2 antibodies) were determined by a double-antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) and immunoblot analysis. We found that 40% of HIV+ and 23% of HIV– homosexual men were positive for the gG-2 antibodies, what is 3.6 and 2.1 times higher incidence, respectively, than that in the control heterosexual population (Bystrická et al., Acta Virol. 42, 319–324, 1998). Identification of individuals infected with genital herpes among HIV+ and HIV– homosexual men should be succeeded by antiviral therapy in order to prevent transmission of HSV-2 and HIV as well in this community.
Key words: HSV-2; HSV-2 antibodies; HIV; glycoprotein G-2; immunoblot
analysis; ELISA; homosexual men
Acta virologica 44: 163 – 167, 2000
E. BAUMEISTER, M.M. SIQUEIRA, V. SAVY, F. FRIEDRICH
Servicio de Virosis Respiratorias, Departamento de Virologia, INEI-ANLIS ”C.G.
Malbran”, Buenos Aires, Argentina;
Departamento de Virologia, Instituto Oswaldo Cruz, FIOCRUZ, 21040-360 Rio de Janeiro, RJ,
Brazil
Summary. – This study reports on genomic characterization of six measles virus (MV) isolates obtained from a measles epidemic in Argentina in 1998. Reverse transcription–polymerase chain reaction (RT-PCR) and nucleotide sequence analysis of the carboxyl-terminal region of the nucleoprotein (N) gene of these isolates classified all of them as wild type MV of D6 genotype. MVs of D6 genotype with identical nucleotide sequences in the region analyzed were also identified during the 1997 measles epidemic in Brazil and the 1999 measles outbreak in Uruguai. These results suggest that the MVs associated with the 1998 measles epidemic in Argentina might have originated from Brazil. As the D6 genotype is also widely distributed in Europe, it is possible that this genotype was brought to South America from Europe.
Key words: measles virus; polymerase chain reaction; nucleotide sequencing;
molecular epidemiology; Argentina
Acta virologica 44: 169 – 174, 2000
T. TALLO, M. LAPPALAINEN, V. TEFANOVA, L. PRIIMÄGI
Department of Virology, Estonian Institute of Experimental and Clinical Medicine, Hiiu
42, 11619 Tallinn, Estonia;
Department of Virology, Helsinki University Central Hospital/Haartman Institute, Helsinki,
Finland
Summary. – Distribution of hepatitis C virus (HCV) genotypes among 30 patients with chronic liver diseases and antibodies to HCV (anti-HCV) was investigated. Sera of these patients were analyzed for HCV genotype by reverse transcription–polymerase chain reaction (RT-PCR) and restriction fragment length polymorphism (RFLP) analysis of the 5´-non-coding region (5´-NCR) of the virus genome and for HCV serotype by detecting antibodies to HCV NS4 peptides by enzyme-linked immunosorbent assay (ELISA). The following distribution of genotypes was found: genotypes 1b in 32.0%, 3a in 20.0%, 2a in 12.0% and 1a/b (double infection) in 28.0%. The results of serotyping were interpretable in 92.0% and concordant with those of genotyping in 80.0% of the patients. In Northern Estonia, the genotypes 1b and 3a seem to be most common in chronically infected patients. Serotyping is an generally available and cheap assay and can be performed in most diagnostic laboratories in comparison to genotyping. However, genotyping is a more sensitive and more specific assay.
Key words: hepatitis C virus; genotyping; serotyping
Acta virologica 44: 175 – 178, 2000
A.T. OWOLABI, E. PROLL
Department of Biological Sciences, University of Calabar, P.M.B. 1115, Calabar,
Nigeria;
Institute for Resistance Research and Pathogen Diagnostics, Aschersleben, Germany
Summary. – An attempt was made to distinguish between celosia mosaic virus (CIMV) and asparagus virus 1 (AV-1) based on biological properties, which hitherto was obscured from serological data from previous work. The host range of AV-1 was found to be a subset of that of CIMV and AV-1was transmitted by the aphid Myzus persicae which, on the other hand, did not transmit CIMV. No evidence of cross-protection was obtained between these two viruses.
Key words: celosia mosaic virus; asparagus virus-1; host range; insects;
transmission; cross-protection
Acta virologica 44: 179 – 182, 2000
K. KANDA, T. KAYASHIMA, F. KATO, A. MURATA
Institute of Applied Microbiology, Department of Applied Biological Sciences, Saga University, Saga 840-8502, Japan
Summary. – Induction of a plasmid-integrative J7W-1-related phage in Bacillus thuringiensis serovar indiana by ethidium bromide was influenced by the temperature at which the host cells were cultured. Under optimal growth conditions, the maximum titer of the phage produced by the serovar indiana reached 1.2 x 106 PFU/ml at 37°C while at 27°C it was lower by an order of magnitude (1.3 x 105 PFU/ml). The temperature-sensitive period was estimated to occur early during the phage induction. However, the temperature effect observed with the serovar indiana did not occur with the serovar israelensis. In the latter case, the phage induction was the same at 37°C or 27°C. Thus we assume that the temperaturesensitive phage induction observed with the serovar indiana as host was not a phenomenon caused by the phage genome but rather by product(s) encoded by certain host gene(s).
Key words: Bacillus thuringiensis; plasmid-integrative phage; J7W-1-related
phage; ethidium bromide induction; temperature sensitivity
Acta virologica 44: 183 – 187, 2000
K. KANDA, Y. TAKADA, F. KAWASAKI, F. KATO, A. MURATA
Institute of Applied Microbiology, Department of Applied Biological Sciences, Faculty of Agriculture, Saga University, Saga 840-8502, Japan
Summary. – Bacillus thuringiensis serovar israelensis, a bacterium which possesses plasmid transfer ability after mating, has been lysogenized by plasmid integrative phage J7W-1. The induction of phage in this J7W-1 lysogen was observed after mating with phage-insensitive strains, such as B. thuringiensis serovar thuringiensis, B. cereus and B. subtilis, as well as the phage-sensitive strain serovar israelensis. The phage induction was not observed after mating with B. thuringiensis strains AF101, serovar dendrolimus and serovar indiana. Because these strains are naturally associated with J7W-1 or its related phage, the data strongly suggest a constitutive expression of the repressor encoded by the prophage in these strains. However, the phage induction was observed in B. thuringiensis serovar aizawai, although it contained the J7W-1 DNA homologous region(s).
Key words: plasmid integrative prophage; phage induction by mating; Bacillus
thuringiensis serovar israelensis
Acta virologica 44: 189 – 192, 2000
C.T. TIEMESSEN, D.J. MARTIN
AIDS Virus Research Unit, National Institute for Virology and Department of Virology, University of the Witwatersrand, Johannesburg, Republic of South Africa
Summary. – Spontaneous secretion of interleukin 8 (IL-8) was higher in latently infected U1 cells than in acutely infected or uninfected parental U937 cells. However, the induction of IL-8 by various cytokines (IL-1alpha, TNF- alpha, IL-6, TNF-beta, GM-CSF, IFN-gamma) was significantly reduced in U1 cells. Cytokine modulation of IL-8 production in U937 cells acutely infected with a T cell-tropic strain (IIIB) or monocytotropic strain (ADA) of human immunodeficiency virus 1 (HIV-1) (HIV-1IIIB and HIV-1ADA) was variable and showed strain-specific differences. The obtained results showed that the in vitro induction of IL-8 is impaired in promonocytic cells latently infected with HIV-1 and is differently modulated under acute conditions of infection depending on the IL-8 inducing cytokine and on the infecting virus strain.
Key words: interleukin-8; HIV-1; promonocytic cells; chronic and acute
infection
Acta virologica 44: 193 – 198, 2000
N. IMAI, W. KANG, K. IWABUCHI, K. SATO, S. MAEDA
Laboratory of Molecular Entomology and Baculovirology, RIKEN, 2-1 Hirosawa, Wako
351-0198, Japan;
Department of Plant Protection, Tokyo University of Agriculture and Technology, Fuchu,
Japan;
Core Research for Evolutional Science and Technology Project, Japan Science and Technology
Corporation, Kawaguchi, Japan
Summary. – Baculovirus IE-2 protein is one of well-known transactivators. In this report, we demonstrate that Bombyx mori nucleopolyhedrovirus (BmNPV) IE-2 interacts with itself. Several clones were obtained from a yeast two-hybrid screening system using IE-2 as bait and were found to encode IE-2 protein. Nucleotide sequencing of these clones showed that they contained C-terminal regions in common. Further analyses suggest that BmNPV IE-2 protein interacts with itself through 80 amino acid residues of coiled-coil domain in C-terminus.
Key words: Bombyx mori nucleopolyhedrovirus ; IE-2 transactivator; yeast two-hybrid
system; coiled-coil domain
Acta virologica 44: 199 – 202, 2000
M. KRZYŻOWSKA, A. SCHOLLENBERGER, M.G. NIEMIAŁTOWSKI
Division of Immunology, Department of Microbiology and Immunology, Faculty of Veterinary Medicine, Warsaw Agricultural University, Grochowska 272, 03-849 Warsaw, Poland
Summary. – Eighty to hundred percent of patients positive for human immunodefficiency viruses 1 or 2 (HIV) may develop opportunistic viral infections. According to the National Institute of Health data, only in the USA the HIV patients are positive also for human cytomegalovirus (HCMV) in 25–40%, varicella-zoster virus (VZV) in 10%, herpes simplex viruses 1 and 2 (HSV-1 and HSV-2), and Kaposi’s sarcoma-associated herpesvirus (human herpesvirus 8, KSHV, HHV-8) in 20%. HIV and herpesviruses express numerous different proteins that are able to influence interactions between the host and virus. One of the most interesting regulatory phenomenon is apoptosis which could play a significant role during both specific and non-specific antiviral response and latency. Apoptosis is an ordered cascade of precisely regulated enzymatic reactions which may be modulated or even controlled by viruses. Dramatic changes which occur during infection and which are exerted by HIV and certain herpesviruses on the mechanism of apoptosis may contribute to the pathogenesis of acquired immunodeficiency syndrome (AIDS).
Key words: apoptosis; herpesviruses; HIV
Acta virologica 44: 203 – 210, 2000
J. MISTRÍKOVÁ, H. RAŠLOVÁ, M. MRMUSOVÁ, M. KÚDELOVÁ
Department of Microbiology and Virology, Faculty of Natural Sciences, Comenius
University, Mlynská dolina B2, 842 15 Bratislava;
Institute of Virology, Slovak Academy of Sciences, Bratislava, Slovak Republic
Summary. – In 1976, within a project on isolation of herpesviruses from small rodents in former Czechoslovakia, the mouse herpesvirus strain 68 (MHV-68) was isolated (Blaškovič et al., 1980). This virus was accepted by The International Committee on Taxonomy of Viruses (ICTV) as a new, so far unassigned species (member) of the Gammaherpesvirinae subfamily of the Herpesviridae family (Murphy et al., 1995). Besides MHV-68, four more isolates (MHV-60, MHV-72, MHV-76, and MHV-78) similar to MHV-68 were obtained in that field experiment in Slovakia. Later, three more isolates (MHV-Šumava from Bohemia and MHV-4556 and MHV-5682 from Slovakia) were obtained in other field experiments. All these isolates are in some properties similar but in others different from each other. Nevertheless, as their comparative genome sequence analysis is not yet available, we propose at present to regard all the abovementioned isolates as different isolates of the same virus and strain, i.e. MHV-68. It is not excluded that a more detailed characterization of these isolates in the future will lead to proposals of designating some of these isolates as new strains of the virus of concern. This review summarizes the up to date knowledge of various biological and physico-chemical properties of MHV-68. At least three isolates, MHV-68, MHV-72 and MHV-Šumava seem to be involved in malignant neoplasm development in mice. It should be stressed that the pathogenesis of the induced lymphoproliferative disease in mice is similar to that caused by Epstein-Barr virus (EBV) in humans.
Key words: mouse herpesvirus strain 68 (MHV-68); murine gammaherpesvirus;
MHV-60, MHV-72, MHV-76, MHV-78, MHV-4556, MHV-5682, MHV-Šumava; characterization;
Epstein-Barr virus; lymphoproliferative disease; pathogenesis
Acta virologica 44: 211 – 226, 2000
D. JANUSZKIEWICZ, J. WYSOCKI, J. NOWAK
Institute of Human Genetics, Polish Academy of Sciences, Strzeszyńska 32, 60-479
Poznań, Poland;
Karol Marcinkowski University of Medical Sciences, Poznań, Poland;
Department of Medical Diagnostics, Poznań, Poland
Key words: HCV; genotypes; hemophiliacs
Acta virologica 44: 227 – 228, 2000
R. PESHEV, L. CHRISTOVA
Central Veterinary Research Institute, 15a P. Slaveikov Blvd., 1606 Sofia, Bulgaria;
Institute of Biophysics, Bulgarian Academy of Sciences, Sofia, Bulgaria
Acta virologica 44: 229 – 230, 2000