Electronic Library of Scientific Literature Electronic Library of Scientific Literature
Volume 44 / April 2000 / number 2
M.L. NOGUEIRA, J.B. AMORIM, J.G. OLIVEIRA, C.A. BONJARDIM, P.C.P. FERREIRA, E.G. KROON
Laboratório de Vírus, Departamento de Microbiologia, Instituto de Ciencias Biológicas, Universidade Federal de Minas Gerais Av. Antonio Carlos, 6627 Belo Horizonte, MG, 31270-901 Brazil
Summary. - We compared two polymerase chain reaction (PCR) assays (simple and multiplex) and viral isolation to detect herpes simplex virus 1 (HSV-1) and varicella-zoster virus (VZV) in 15 clinical specimens from 13 patients with mucocutaneous herpetic infections. HSV-1 or VZV DNA was detected in 13 specimens by simple PCRs (HSV-1 or VZV PCR) and in 12 specimens by multiplex PCR. On the other hand, viral isolation was positive for 9 specimens only. The PCR protocols used in this study are not only more sensitive and faster than the traditional viral isolation and conventional PCR protocols but also can distinguish rapidly HSV-1 from VZV. We propose the PCRs described here for rapid and precise identification of etiological agents of mucocutaneous herpetic infections.
Key words: HSV-1; VZV; PCR; virus isolation; mucocutaneous herpetic
infections; diagnostics
Acta virologica 44: 61 - 65, 2000
I.D. GARG, V. HEGDE, S.M.P. KHURANA
Division of Plant Pathology, Central Potato Research Institute, Shimla 171 001, H.P. India
Summary. - Electron microscopic studies on the stability of immunosorbed (trapped) virions of potato viruses X, S and Yo (PVX, PVS and PVYo) revealed disintegration and dislodging of PVYo virions upon incubation with (1) antisera to PVX, PVS, or both diluted in saline, (2) 0.86% NaCl (saline) or 0.1 mol/l CaCl2 but not with 0.1 mol/l CaSO4 or 0.1 mol/l MgSO4. PVX virions, on the other hand, showed partial dislodging upon incubation with an antiserum to PVS diluted in saline, but complete disintegration and dislodging with saline. 0.1 mol/l CaCl2 caused partial dislodging while MgCl2, CaSO4 or MgSO4 (all 0.1 mol/l) had no apparent adverse effect. PVS virions were not affected by saline, CaCl2, MgCl2, CaSO4 or MgSO4 (all 0.1 mol/l) and were only partially dislodged by antisera to PVX or PVYo. Disintegration and/or dislodging of the PVX and PVYo virions was prevented when (1) they were fixed with glutaraldehyde prior to incubation or (2) the virus extract contained bovine serum albumin (BSA) or (3) heterologous antisera were diluted in 0.1 mol/l phosphate buffer (PB) before use except the PVS antiserum which still caused disintegration and dislodging of PVYo virions. Prior fixation of virions prevented their disruption and dislodging by saline only in the case of PVYo but not PVX. On the other hand, BSA reverted the adverse effect of saline but not that of the PVS antiserum on PVYo virions. The results presented here suggest (1) a disruptive effect of Cl' on PVX and PVYo virions particularly when it was associated with Na+ and (2) an interaction between the immunosorbed virions of PVX or PVYo and the antiserum to PVS.
Key words: PVX; PVS; PVYo; immunosorbed virions; stability
Acta virologica 44: 67 - 72, 2000
L. NIKOLAEVA, A.S. GALABOV
Department of Virology, The Stephan Angeloff Institute of Microbiology, Bulgarian Academy of Sciences, 26 G. Bonchev Str., 1113 Sofia, Bulgaria
Summary. - Effects of enviroxime and disoxaril, inhibitors of replication of some picornaviruses with known mechanisms of action, alone or in combination, on replication of coxsackievirus B1 (CVB1) in FL cells and on experimental CVB1 infection in newborn mice were tested. The combination of enviroxime and disoxaril resulted in vitro in a synergistic interaction. Both compounds were administered in vivo, alone or in combination, daily by subcutaneous (s.c.) route since the day of virus inoculation till the 5th day post inoculation (p.i.). Our findings about the in vivo antiviral effects of the individual compounds correlated with those of other authors, i.e. disoxaril significantly reduced the virus-induced death (the minimum 50% effective dose (ED50) was 12.5 mg/kg; P = 0.0037), while enviroxime was not effective even when applied at a dose as high as 100 mg/kg (P = 0.264). However, when both the substances were combined, the same protective effect was achieved with concentrations of disoxaril two to four times lower than those of the drug administered alone. In this way a higher selectivity ratio was achieved. Namely, the combination of 50 mg/kg enviroxime and 3.125-6.25 mg/kg disoxaril was synergistic. Along with reduction in mortality a marked delay in the course of the disease was observed.
Key words: coxsackievirus B1; enviroxime; disoxaril; antivirals; in vivo;
synergism
Acta virologica 44: 73 - 78, 2000
V. UCHAIKIN, T. CHEREDNICHENKO, V. MALINOVSKAYA, T. ORLOVA, O. KOVALEV, F. KHARLAMOVA, G. CHAPLYGINA, V. KONEV, F. VORONINA, N. DELENIAN
Russian State Medical University, Moscow, Russia;
Gamaleya Research Institute for Epidemiology and Microbiology, Russian Academy of Medical
Sciences, Gamaleya 18, Moscow 123098, Russia
Summary. - A new antiviral and immunomodulating preparation Viferon, produced as rectal suppositories containing recombinant alpha-2b-interferon (IFN) and antioxidants, was used in complex therapy of viral chronic hepatitides B and C (ChHB and ChHC) in children. Results of our investigation showed a high efficiency of Viferon. Viferon was found to suppress replication of hepatotropic viruses and to decrease activity of the pathologic process in the liver of children with ChHB and ChHC. After a Viferon treatment with daily doses of (1-2) x 106 IU of IFN (3.0 x 106 IU/m2) primary remission was registered in 78% of patients with ChHB and in 44% of patients with ChHC, while lasting remission was found in 82% of ChHB and in 33% of ChHC patients. Thus, a more marked effect was observed with ChHB, in which 3.0 x 106 IU/m2 was the optimal daily dose for children. Increasing the dose to 5.0 x 106 IU/m2 did not result in rise of the percentage of the remissions. Side effects, which are characteristic for injection of IFN preparations, were never found even after a longterm treatment. Absence of induction of neutralizing antibodies was observed after administration of alpha-2b-IFN, an integral part of Viferon. In pediatrics, the method of rectal administration has advantages over parenteral delivery due to its convenience, non-traumatic character and possibility of use for prolonged periods.
Key words: chronic viral hepatitis; treatment; interferon; Viferon;
suppository
Acta virologica 44: 79 - 83, 2000
A. SANYAL, C.B. GURUMURTHY, R. VENKATARAMANAN, D. HEMADRI, C. TOSH
Central Laboratory, All India Coordinated Research Project on Foot-and-Mouth Disease, Indian Veterinary Research Institute, Mukteswar-Kumaon, Nainital 263 138, U.P., India
Summary. - The nucleotide and deduced amino acid sequences in the amino acid (aa) 130-160 region of VP1 polypeptide of 65 field isolates of foot-and mouth disease virus (FMDV) serotype Asia 1 were determined and the consensus sequences were deduced. Comparison of amino acid sequences revealed conservation of NGK (130-132), TYG (134-136), RGD (142-144), and LPTSF (156-160) motifs and aa 148 (L) while variation was observed at the rest of the region (variability index (VI) of 2.06 to 16.85). Synonymous and non-synonymous mutations at the nucleotide level were well correlated with those of the corresponding amino acids. Comparison of the aa 130-160 sequence of Asia 1 serotype with those of other serotypes of FMDV revealed conservation of aa 135, 148-149, 157 and 160. Amino acids 133-138 and 148-154 were unique for Asia 1 serotype and are presumably responsible for its distinct antigenic nature. The present study revealed that the FMDV isolates of serotype Asia 1 causing outbreaks in India are very much heterogeneous in the aa 130-160 region of VP1.
Key words: FMD virus; serotype Asia 1; Indian isolates; VP1; amino acid
sequence; variability index
Acta virologica 44: 85 - 90, 2000
M. SHINJOH, K. OMOE, N. SAITO, N. MATSUO, K. NEROME
Department of Virology I, National Institute of Infectious Diseases, Tokyo, Japan;
Department of Pediatrics, School of Medicine, Keio University, Tokyo, Japan;
Laboratory of Electron Microscopy, National Institute of Infectious Diseases, Tokyo, Japan
Summary. - To elucidate epidemiological interference between respiratory syncytial (RSV) and influenza viruses, the influence of influenza A (H1N1) virus on the growth of RSV was examined. Although RSV grew in MDCK cells, coinfection with influenza A virus led to a reduction of progeny RSV. The degree of growth interference depended on the time of infection with influenza A virus post infection (p.i.) with RSV. In fact, infection with influenza A virus 12 hrs p.i. with RSV did not influence growth of the latter virus. On the contrary, growth suppression of influenza A virus by RSV was observed when the coinfection began at the later stages of RSV infection. Suppression of the growth of RSV by influenza A infection was further demonstrated at the level of viral protein synthesis. An indirect immunofluorescence (IF) test revealed that a large proportion of infected cells synthesized both RSV and influenza A virus antigens. Scanning electron microscopic (SEM) examination demonstrated that influenza A and RSV virions possessing surface antigens specific for each virus were selectively released from dually infected cells. In the present study, we proved for the first time that the growth of RSV is blocked by competitive infection with influenza A virus in a susceptible cell population, competitive protein synthesis and selective budding of RSV and influenza viruses from the same infected cells.
Key words: respiratory syncytial virus; influenza A virus; interference; dual
infection; MDCK cells
Acta virologica 44: 91 - 97, 2000
I. Oravcová, M. Kúdelová, J. Mlčuchová, J. Matis, M. Bystrická, D.F. Westra, S. Welling-Wester, J. Rajčáni
Institute of Virology, Slovak Academy of Sciences, Dúbravská cesta 9,
842 45 Bratislava, Slovak Republic;
DSM Biologics, Hoogkerk, The Netherlands;
Department of Medical Microbiology, University of Groningen, Groningen, The Netherlands
Summary. - Sequences of UL44 genes of strains HSZP, KOS and 17 of herpes simplex virus 1 (HSV-1) were determined and the amino acid sequences of corresponding glycoproteins (gC) were deduced. In comparison with the 17 strain, the HSZP strain showed specific changes in 3 nucleotides and in 2 amino acids (aa 139 and 147, both from Arg to Trp) in the antigenic locus LII. The change at aa 147 was situated within the GAG-binding epitope. In a similar comparison, KOS strain had changes in 3 nucleotides and 3 amino acids (aa 3, 14, and 300). The UL44 genes of HSZP and KOS strains were expressed in insect Sf-21 cells by means of the baculovirus (Bac-to-BacTM) expression system. As shown by immunoblot analysis, both the recombinant baculoviruses (B1-HSZP and B6-KOS) expressed a glycosylated gC, the Mr of which (116 K) was lower than that of gC synthesized in Vero cells (129 K) infected with strains HSZP or KOS. In addition, smaller gC-specific proteins (of apparent Mr of 50-58 K and 98 K) corresponding to a non-glycosylated precursor polypeptide and/or incomplete forms of the partially glycosylated gC were found. When Balb/c mice were immunized with Sf-21 cells expressing gC, the recombinant gC-HSZP represented a more efficient immunogen possibly due to its stronger expression in these cells. The corresponding gC-HSZP antiserum reacted in enzyme-linked immunosorbent assay (ELISA) equally well with HSZP and KOS virion antigens and neutralized HSZP strain at a low titer. Both gC-HSZP and gC-KOS antisera detected the homologous as well as the heterologous gC antigens in Vero cells regardless whether infected with strains HSZP, KOS or 17, revealing the presence of gC from 6 to 16 hrs post infection (p.i.) in the cytoplasm, on the nuclear membrane and at the cell surface.
Key words: HSV-1; HSZP strain; KOS strain; 17 strain; UL44 gene; glycoprotein
C; mutations; immunogenicity; baculovirus expression system
Acta virologica 44: 99 - 108, 2000
F. Friedrich
Departamento de Virologia, Instituto Oswaldo Cruz, FIOCRUZ, 21040 360 Rio de Janeiro, RJ, Brazil
Summary. - The oral poliovirus vaccine (OPV) has been effectively used in the control of poliomyelitis and in the eradication of wild polioviruses. Although there are many advantages in using attenuated OPV strains in the campaign to eradicate poliomyelitis, several studies have demonstrated that there are some disadvantages such as (a) excretion by vaccines of OPV-derived polioviruses with genomic modifications known to increase the neurovirulence, (b) appearance of vaccine-associated paralytic poliomyelitis (VAPP) and other adverse effects in vaccinees, (c) occurrence of persistent infections caused by OPV-derived strains in immunodeficient patients with VAPP, (d) transmission of OPV-derived polioviruses to susceptible individuals which develop VAPP, and (e) detection of OPV-derived polioviruses in the environment, which could be a source of infection for humans in the future. Different studies indicate that it is important to consider the possibility of persistent infections and excretion of OPV-derived polioviruses for long periods by humans, and also the survival in the environment of OPV-derived polioviruses excreted by humans, which could be transmitted and circulate in a non-immune population after stopping poliovirus vaccination. The findings reported here may have important implications for global poliomyelitis eradication initiative and indicate that surveillance of OPV-derived strains will also be important in the final step of eradication of poliomyelitis from the planet.
Key words: poliovirus; poliomyelitis; oral poliovirus vaccine, poliomyelitis
eradication, vaccine-derived polioviruses, mutation, recombination, deletion
Acta virologica 44: 109 - 117, 2000
K. SUBI
Department of Virology, Institute of Experimental and Clinical Medicine, Hiiu 42, 11619 Tallinn, Estonia
Acta virologica 44: 119 - 120, 2000
I. VANČOVÁ, E. MARTENS, K. VANDENBROECK, E. KONTSEKOVÁ, A. BILLIAU, P. KONTSEK
Institute of Virology, Slovak Academy of Sciences, Bratislava, Slovak Republic;
Rega Institute, Catholic University Leuven, Leuven, Belgium;
Department of Microbiology and Virology, Faculty of Sciences, Comenius University,
Bratislava;
Institute of Neuroimmunology, Slovak Academy of Sciences, Bratislava
Key words: porcine interferon gamma; antigenic structure; monoclonal antibodies;
pH 2 treatment
Acta virologica 44: 121 - 122, 2000