Electronic Library of Scientific Literature Electronic Library of Scientific Literature
Volume 43 / October 1999 / number 5
K. YOSHIIE, S. MATAYOSHI, T. FUJIMURA, N. MAENO, H. ODA
Department of Bacteriology, Faculty of Medicine, Kagoshima University, Sakuragaoka 8-35-1, Kagoshima 890-8520, Japan
Summary. – We compared in vitro sensitivities to Coxiella burnetii of alveolar macrophages, derived from mice sensitive and resistant to C. burnetii, respectively, and examined the role of nitric oxide (NO) in the C. burnetii infection. Alveolar macrophages of sensitive A/J mice showed a larger population of C. burnetii antigen-positive cells than those of resistant C57BL/6 mice. C. burnetii induced NO production in alveolar macrophages, but N-methyl-L-arginine and sodium nitroprusside (SNP), NO inhibitor and donor, respectively, did not inhibit the infection. Thus the NO induction seems to be independent of the cell defense mechanism against the C. burnetii infection.
Key words: Coxiella burnetii; alveolar macrophage; nitric oxide; in vitro
sensitivity
Acta virologica 43: 273 – 278, 1999
C. CHASTEL, L. CHANDLER, F. LE GOFF, O. CHASTEL, R. TESH, R. SHOPE
Laboratoire de Virologie, Faculté de Médecine, F 29285 Brest Cédex, France;
WHO Collaborating Center for Tropical Diseases, University of Texas Medical Branch at
Galveston, Galveston, TX, USA;
CEBC/CNRS, Villiers-en-Bois, France
Summary. – A new phlebovirus (Bunyaviridae family, Phlebovirus genus), provisionally designed Chizé virus, was isolated from a nymph of Ixodes (Trichotoixodes) frontalis collected on a wren (Troglodytes troglodytes) found dead in the Chizé forest, western France. Chizé virus produced a lethal encephalitis in one-day-old mice and cytopathic effect (CPE) in Vero cells. Extracellular particles with a mean diameter of 105 nm with surface spikes characteristic of Uukuniemi (UUK) serogroup viruses were observed in Vero cells. Chizé virus reacted in complement-fixation test with several UUK serogroup viruses but was readily distinguished from all registered viruses in the serogroup. I. frontalis is highly specific for birds and unlikely to transmit Chizé virus to humans or domestic animals; the pathogenicity of the new virus to wild birds remains to be clarified.
Key words: Chizé virus; phlebovirus; bunyavirus, Ixodes frontalis;
Trichotoixodes frontalis; wren; Troglodytes troglodytes; France
Acta virologica 43: 279 – 283, 1999
A. TAHA, X. LÉRY, J. GIANNOTTI
Entomovirology Laboratory, ORSTOM, P.O.B. 26, Giza code 12211, Cairo, Egypt
Summary. – In analyzing populations of non-infected potato tuber moth (PTM) Phthorimaea operculella, using a total DNA probe from Phthorimaea operculella granulovirus (PhopGV), false positive reactions were obtained indicating homology between cellular and viral DNAs. Using a cloned 2.1 kbp fragment of PhopGV DNA, a specific digoxigenin-labeled probe was developed. This fragment did not show homology using both dot and Southern blot hybridization with either the genome of the larvae or genomes of the cell lines derived from the insect. The PhopGV-specific DNA probe detected as little as 1 ng, while the total DNA probe could detect even 35 pg of purified viral DNA. The 2.1 kb probe was highly specific for PhopGV. It gave negative results with two other granuloviruses isolated on Sesamia cretica and Spodoptera littoralis. The availability of a PhopGV-specific probe is an important prerequisite of detection of early stages of virus infection both in vivo on P. operculella larvae and in vitro on established P. operculella cell lines.
Key words: potato tuber moth; Phthorimaea operculella; granulovirus; specific
DNA probe; virus detection; cell line
Acta virologica 43: 285 – 289, 1999
N.E. BISHOP
Hepatitis Research Unit, Macfarlane Burnet Centre for Medical Research, P.O. Box 254, Fairfield, Victoria 3078, Australia
Summary. – Cleavage of VP0 to VP2 via intramolecular scission is known as the viral maturation cleavage, as VP0 is found in immature particles, whilst VP2 is found in mature particles. The effect of low pH on the kinetics of hepatitis A virus (HAV) capsid protein VP0 cleavage in provirions was examined by Western blot analysis. VP0 scission was found to be dramatically enhanced under acidic conditions, similar to those encountered on entry of virus particles into the cell via endocytosis. The cleavage of VP0 to VP2 led to an increase in the specific infectivity of viral particles, indicating that mature virions are more infectious than immature provirions. The data are consistent with a model where conformational changes induced by low pH aid scission of VP0, and the increase in kinetics of VP0 cleavage may have relevance for viral uncoating, as only mature HAV particles are thought capable of uncoating within the host cell.
Key words: picornavirus; uncoating; provirion; VP0; VP2; maturation cleavage
Acta virologica 43: 291 – 296, 1999
T.V. S. RAO, POONAM MALIK, D. ASGOLA
Division of Virology, Indian Veterinary Research Institute Campus, Mukteswar-Kumaon, Nainital 263138, U.P., India
Summary. – A noninfectious soluble antigen fraction of goat poxvirus (GPV) fractionated by ammonium sulfate precipitation was tested for its suitability as coating antigen in an indirect enzyme-linked immunosorbent assay (ELISA). Accordingly, an avidin-biotin ELISA for the detection of GPV antibodies was optimized and evaluated using different groups of serum samples from goats with known or unknown immune status. A cut-off value higher by 60% than A492 reading of control negative sera gave a 91.8% specificity and a 94.1% sensitivity for the assay. Out of 90 goat pox-suspect sera obtained from the field, only 2 (2.2%) were found positive in the counter immunoelectrophoresis (CIE) test, which is so far the routinely used diagnostic test for goat pox, while 58 (64.4%) were positive in the avidin-biotin ELISA. The McNemar’s analysis of these data showed that the avidin-biotin ELISA was significantly more efficient than the CIE test for the detection of GPV antibodies in goat sera.
Key words: antibody; avidin-biotin ELISA; counter immunoelectrophoresis;
goat poxvirus; noninfectious diagnostics; orf; soluble antigen
Acta virologica 43: 297 – 301, 1999
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. – To assess the possible interactions among picornavirus replication inhibitors, inhibitory effects of dual combinations of enviroxime, disoxaril, arildone, S-7, guanidine, PTU-23, and HBB on poliovirus type 1 (Mahoney) replication in FL cells were tested. Beforehand, the 50% inhibitory concentration (IC50) in the plaque inhibition test was been determined for each individual compound, i.e. enviroxime – 0.2 micromol/l, disoxaril – 0.3 micromol/l, arildone – 2.7 micromol/l, S-7 – 100 micromol/l, guanidine – 200 micromol/l, PTU-23 – 200 micromol/l, and HBB – 300 micromol/l. Each of the dual combinations, in which enviroxime or HBB was one of the partners, showed synergistic or additive effects. Combining disoxaril with enviroxime, HBB or PTU-23 resulted in synergism, while combining it with guanidine, S-7 or arildone led to antagonism. Arildone showed additive or synergistic effects when combined with enviroxime, HBB and PTU-23, and antagonistic ones when combined with disoxaril, S-7 or guanidine. All dual combinations of PTU-23 were synergistic with the exception of the pair of PTU-23 + guanidine that was antagonistic. Guanidine had additive to synergistic interactions with HBB or enviroxime but antagonistic ones with disoxaril, arildone and PTU-23. Guanidine or PTU-23 when combined with S-7 showed an unusual effect – synergistic one with an antagonistic zone. The combinations of S-7 with enviroxime or HBB were synergistic but those with disoxaril or arildone were antagonistic. Research on interactions of picornavirus replication inhibitors could possibly contribute to the development of efficient chemotherapy of infectious diseases caused by picornaviruses as well as to the better understanding of the mode of action of those inhibitors.
Key words: antivirals; poliovirus; synergy; antagonism; mechanism of action
Acta virologica 43: 303 – 311, 1999
V. REPKA
Laboratory of Molecular Biology and Virology, Research Institute of Viticulture and Enology, Matúškova 25, 833 11 Bratislava, Slovak Republic
Summary. – Three different molecular forms, isoforms, of the major virus-inducible anionic peroxidase (PRX) of cucumber (Cucumis sativus L.) were purified to homogeneity from crude extracts of hypersensitively reacting cotyledons and subjected to proteolysis with five exogenous endoproteinases. The PRX isoforms were fully resistant to degradation by trypsin and chymotrypsin even though at a prolonged incubation. Partial proteolysis with pepsin yielded peptides which were similar in size and serological properties. When papain was used, the peptides released from PRX1 isoform differed both in size and number but not serologically from the peptides released from isoforms PRX2 and PRX3 confirming similar primary structure of polypeptide chains. PRX3 was the only substrate giving a peptide map after incubation with protease K. Under experimental conditions used in this work, PRX1 and PRX2 were degraded completely with protease K. These results indicate that PRX1, PRX2, and PRX3 contain similar antigenic determinants and indicate very similar but not identical primary structures. Several practical implications of the present study are also mentioned.
Key words: cucumber; tobacco necrosis virus; proteolysis; electrophoresis; Western
blot analysis
Acta virologica 43: 313 – 319, 1999
P.K. GUPTA, M. SAINI, L.K. GUPTA, S.K. GARG
National Biotechnology Centre and
Division of Biochemistry and Food Science, Indian Veterinary Research Institute,
Izatnagar, 243 122, U.P., India;
Department of Veterinary Microbiology, G.B. Pant University of Agriculture and Technology,
Pantnagar, U.P., India
Summary. – Bovine peripheral blood mononuclear cells (PBMCs) and monocytes were stimulated with bovine herpesvirus 1 (BHV-1), LPS and concanavalin A (Con A) to produce L-arginine-dependent nitric oxide (NO) in vitro. NO was detected as early as 12 hrs and up to 72 hrs post stimulation (p.s.). The NO from lipopolysacharide (LPS)-stimulated PBMCs and monocytes was found to exhibit antiviral effect against BHV-1. The anti-BHV-1 effect was inhibited with Nw-methyl L-arginine indicating involvement of inducible NO synthase (iNOS) in NO production.
Key words: bovine herpesvirus 1; monocytes; NO; NO synthase; peripheral blood
mononuclear cell
Acta virologica 43: 321 – 324, 1999
V. ARRUNATEGUI-CORREA, S. BALTATZIS, C.S. FOSTER
Department of Microbiology and Immunology, Vanderbilt University, Nashville, TN, USA;
University of Athens, Athens, Greece;
Hilles Immunology and Rhoads Molecular Immunology Laboratories, Massachussetts Eye and Ear
Infirmary, Harvard Medical School, 243 Charles St., Boston, MA 02114, USA
Summary. – Experimental corneal infection with herpes simplex virus 1 (HSV-1) resulted in 11–21 days in herpes simplex keratitis (HSK) in C.Al-20 but not C.B-17 strain of BALB/c Igh-1-disparate mice. Formation of mRNAs of various pro-inflammatory cytokines was analyzed in corneas and draining lymph nodes (LNs) of HSK-susceptible C.Al-20 and HSK-resistant C.B-17 mice following HSV-1 corneal inoculation by reverse transcription-polymerase chain reaction (RT-PCR) and Southern blot analysis. Transcripts for interleukin (IL)-2 and interferon (IFN)-gamma were expressed in LNs of susceptible but not resistant mice. The level of IL-6 expression in the cornea correlated with the severity of keratitis in susceptible mice, being evident at days 4 and 14 after virus inoculation and thus showing a biphasic response. Resistant mice did not develop HSK and did not express IL-6. The IL-1beta and IL-4 gene transcription began early (day 7) in the corneas of resistant mice and then ceased, while in the corneas of susceptible mice, it began later (day 11). Taken together, these results indicate that IL-1beta, IL-4, IL-6, and IL-7 participate in the local inflammatory response in HSK.
Key words: herpes simplex virus 1; herpes simplex keratitis; cytokines; gene
expression; reverse transcription; polymerase chain reaction
Acta virologica 43: 325 – 330, 1999
R. Woessner, M.T. Grauer, A. Haass, B. Gaertner, G. Holzer, D. Mueller-reiland, N. Mueller-Lantzsch, J. Treib
Department of Neurology, University Hospital of the Saarland, D-66421 Homburg/Saar,
Germany;
Department of Virology, University of the Saarland, Homburg/Saar, Germany
Summary. – The goal of the present study was to investigate whether a direct association exists between false-positive recognition of IgG antibodies and inflammatory changes in the central nervous system (CNS) and whether inflammatory diseases of the CNS affect the specificity of the enzyme-linked immunosorbent assay (ELISA) of tick-borne encephalitis (TBE) virus. A group of patients (1,815), treated in the Department of Neurology, University Hospital of the Saarland, Homburg/Saar, Germany, were tested for TBE IgG antibodies by ELISA. Several subgroups of patients with and without inflammatory changes in the CSF as well as patients with and without confirmed multiple sclerosis (MS) were investigated. Overall, 4.5% of all the 1,815 patients and 4.8% of the patients with inflammatory changes in the CSF but without MS had TBE IgG antibodies. In the subgroup with inflammatory changes in the CSF and MS, 4.4% of the patients were TBE IgG-positive. In the subgroup without inflammatory changes in the CSF, 3.8% of the patients without MS were TBE IgG-positive and 4.9% of the patients with MS were TBE IgG-positive. The rate of TBE IgG positivity was not significantly different in the subgroups with and without inflammatory changes in the CSF (P = 0.45). The comparison of the subgroups with and without MS showed no significant difference in the TBE IgG titer (P = 0.83) as well. This indicates that the specificity of the ELISA was affected neither by inflammatory changes in the CSF nor by MS.
Key words: tick-borne encephalitis; multiple sclerosis; inflammatory CNS
disease
Acta virologica 43: 331 – 333, 1999
J. ŠPAK, D. KUBELKOVÁ
Department of Plant Virology, Institute of Plant Molecular Biology, Academy of Sciences of the Czech Republic, Branišovská 31, 37005 České Budějovice, Czech Republic
Key words: raspberry bushy dwarf virus; virus strain; grafting; ELISA; Czech
Republic
Acta virologica 43: 335 – 336, 1999