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Volume 47 / 2003 / number 2
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Articles
Short Communication
Letter-to-the Editors
YADAV, P., SHOUCHE, Y.S., MUNOT, H.P., MISHRA, A.C., & MOURYA, D.T.: Genotyping of Chikungunya virus isolates from India during 1963–2000 by reverse transcription –polymerase chain reaction
TOMAN, R., HUSSEIN, A., SLABÁ, K., & ŠKULTÉTY, Ľ.: Further structural characteristics of the lipopolysaccharide from Coxiella burnetii strain Nine Mile in low virulent phase II
J. SCHMIDT-BRAUNS*
Max-Planck-Institute for Marine Microbiology, Celsiusstr. 1, 28359 Bremen, Germany
Received January 17, 2003; accepted April 2, 2003
Summary. – Some picornaviruses
might use the general increase of ionic strength in the host cell that
occurs successively after infection to induce shutoff of host protein
synthesis and to stimulate viral protein synthesis. In order to
investigate this discrimination mode on a molecular level, in vitro
experiments under different salt conditions comparing the Foot-and-mouth
disease virus (FMDV) internal ribosome entry site (IRES)-dependent
translation with the translation via the classical scanning mechanism
were performed. For classical mRNA optimum concentrations of all
investigated salts ranged between 70 and 100 mmol/l. However, for FMDV
IRES-dependent translation the optima depended strongly on the anion
used. While acetates caused only a weak stimulation of translation
efficiency with maxima ranging between 150 and 180 mmol/l, chlorides
lead to a strong stimulation with maxima ranging between 120 and
150 mmol/l. Competition experiments revealed that the concentration of
chlorides had a greater influence on the discrimination between
cellular and viral RNA translation than the total ionic strength. Taken
together, the data support a model in which a specific
increase in the chloride concentration rather than a general
increase in the ionic strength is responsible for the shutoff effect
induced by some picornaviruses.
Key words: in vitro translation; initiation; elongation;
picornavirus; shutoff; IRES
Acta virologica 47: 65 – 72, 2003
M.V. Bais, R.S. Kataria, A.K. Tiwari*, Indervesh, V.V S. Suryanarayana, S.K. Das
National Biotechnology Center, Indian Veterinary Research Institute, Izatnagar, 243122 UP, India
Received November 15, 2002; accepted April 3, 2003
Summary. – Sequence analysis of
genome segment A of an Indian Infectious bursal disease virus (IBDV)
field isolate (KT1/99) revealed total 95 nucleotide substitutions,
resulting in 17 amino acid changes. Of these, five amino acid changes,
namely F60S, T137I, I374V, V519I and E682D were unique to the KT1/99
isolate. The amino acid change P222A and the proposed hot mutation spot
680Y, reported to be present in very virulent IBDV isolates were also
found in KT1/99. This isolate had nucleotide divergence of 1.1% to 4.95%
from the other reported serotype 1 IBDV isolates and 19.6% from serotype
2 strain OH in polyprotein gene sequence, while divergence at amino acid
level was 0.6% to 2.9% and 11.4%, respectively. Based on both nucleotide
and amino acid sequence analysis, KT1/99 was grouped phylogenetically
with the reported Bangladesh isolate BD3/99 in one cluster along with
other reported very virulent isolates in same lineage.
Key words: Infectious bursal disease virus; nucleotide
sequence; amino acid sequence; segment A
Acta virologica 47: 73 – 77, 2003
M. Xiao1,2, J. Chen2, B. Li2*
1Biology Department, College of Life
and Environment Sciences, Shanghai Teachers' University, Shanghai,
200234 P.R. China;
2Key Laboratory for Biodiversity Science and Ecological
Engineering of Ministry of Education,
The Institute of Biodiversity Science, Fudan University, Shanghai,
200433 P.R. China
Received January 28, 2003; accepted April 4, 2003
Summary. – The NS5B gene, cloned
from Classical swine fever virus (CSFV) genome, was expressed in porcine
kidney cells PK-15, natural host of CSFV. In purifying cytoplasmic
extracts from these cells by means of different concentrations of salt,
glycerol and detergent four fractions, namely crude supernatant (SC) and
different purified supernatants (S1, S2 and S3) were obtained. Using
Western blot analysis the NS5B protein was found in all these fractions,
showing that it was soluble in both higher and lower concentrations of
salt, glycerol and detergent. The NS5B protein present in the four
different fractions exhibited RNA-dependent RNA polymerase (RdRp)
activity, but it was unable to complete the whole process of RNA
synthesis. Site-directed mutation analysis showed that Thy216 and Cyt228
were essential for RNA synthesis while Cyt219 was not, suggesting that
CSFV RdRp was template-specific. We conclude that initiation of RNA
synthesis by CSFV RdRp includes also template priming.
Key words: NS5B gene; Classical swine fever virus;
RNA-dependent RNA polymerase
Acta virologica 47: 79 – 85, 2003
O. Kim1*, S.S. Kim2
1USDA-ARS, ADRU, 319 Bustad, WSU,
Pullman, WA 99164, USA;
2Department of Viral Diseases, National Institute of
Health, Seoul 122-701, Republic of Korea
Received February 27, 2003; accepted April 30, 2003
Summary. – Polymerase chain
reaction (PCR) is a powerful technique of detecting Human
herpesvirus 8 (HHV-8), but has a limited sensitivity and
specificity. A new assay of HHV-8 based on combination of PCR with
dot blot hybridization (DBH) was developed and evaluated for its
sensitivity and specificity. An HHV-8-specific primer pair, ORF26out was
used for amplification of target DNA. When the PCR product was detected
visually the limit of detection was 0.1 ng DNA isolated from
HHV-8-infected BC-3 cells. For DBH, the DNA amplified with the primer
pair ORF26in specific for HHV-8 was labeled with digoxigenin (DIG). This
DIG-labeled probe was capable of detecting 1.0 ng of DNA isolated from
HHV-8-infected BC-3 cells. On the other hand, PCR combined with DBH
(PCR/DBH) was more sensitive than PCR or DBH alone and also very
specific. The sensitivity of PCR/DBH was higher than that of PCR and DBH
alone. The PCR/DBH assay can be applied efficiently to confirm the
presence of HHV-8 in clinical samples and to differentiate specifically
HHV-8 infection from other viral infections.
Key words: polymerase chain reaction; dot blot
hybridization; Human herpesvirus 8
Acta virologica 47: 87 – 90, 2003
O. Kim1*, S.J. Yi2
1USDA-ARS, ADRU, 319 Bustad, WSU,
Pullman, WA 99164, USA;
2College of Veterinary Medicine, Kyungpook National
University, Republic of Korea
Received February 12, 2003; accepted May 6, 2003
Summary. – Although many viruses
can induce apoptosis in infected cells, large DNA viruses, such as
poxviruses, herpesviruses and adenoviruses, usually exhibit the ability
to suppress the induction of apoptosis in the infected cells. We
investigated the ability of Human herpesvirus 8 (HHV-8) to protect cells
from apoptosis induced by the virus. HHV-8 has been shown to harbor
genes with anti-apoptotic capacity. However, we demonstrate here that
a lytic replication of HHV-8 resulted in induction of apoptosis
using different techniques to detect apoptosis. Therefore, despite the
presence of anti-apoptotic genes in its genome, HHV-8 could complete its
cycle of productive infection while inducing apoptosis in infected
cells. This finding might have implications for the pathobiology of
HHV-8 and other gamma herpesviruses in vivo.
Key words: apoptosis; gammaherpesvirus; Human herpesvirus 8
Acta virologica 47: 91 – 95, 2003
C.S. LIM1, G. KUMARASINGHE2, V.T.K. CHOW1*
1Programme in Infectious Diseases,
Department of Microbiology, Faculty of Medicine, National University of
Singapore, Kent Ridge 117597, Singapore;
2Department of Laboratory Medicine, National
University Hospital, Kent Ridge, Singapore
Received March 26, 2003; accepted May 14, 2003
Summary. – To study the genetic
variability and molecular epidemiology of Human respiratory syncytial
virus (HRSV) occurring in Singapore, nucleotide sequencing of three
membrane-associated genes (SH, G and F) of four local isolates was
performed. Comparison of their nucleotide and amino acid sequences with
those of the prototype strains A2 (subgroup A) and CH-18537 (subgroup B)
indicated that the Singapore isolates belong to the subgroup A.
Comparison of the Singapore isolates with the reference strain A2 showed
that whereas the G protein was the most divergent with up to 15%
difference, the F and SH proteins showed less diversity of only up to
4%. Each gene exhibited its distinct variable and conserved regions. The
N- and O-glycosylation sites within the G protein of the isolates were
analyzed to ascertain their potential implications on the antigenicity
of the viral glycoprotein. Based on the second variable region of the G
protein, phylogenetic analysis of the Singapore isolates with 91
previously identified genotypes of subgroup A revealed that more
than one genotype (GA2 and GA5) may circulate in the local population at
a given time. This epidemiological study reflects the pattern of
genetic relationships between the HRSV isolates from Singapore to those
from other parts of the world.
Key words: F, G, SH genes/proteins; genotypes; phylogeny;
Human respiratory syncytial virus; sequence analysis
Acta virologica 47: 97 – 104, 2003
G. Raikhy, V. Hallan, S. Kulshrestha, M. Lal Sharma, R. Ram, A.A. Zaidi*
Plant Virology Laboratory, Floriculture Division, Institute of Himalayan Bioresource Technology, CSIR, Palampur, Himachal Pradesh, 176 061, India
Received April 29, 2003; accepted May 21, 2003
Summary. – Incidence of the
Carnation etched ring virus (CERV), the only DNA virus reported to date
on carnation, was investigated by a bioassay using a partially
purified virus as inoculum and then by a double-antibody sandwich
enzyme-linked immunosorbent assay (DAS-ELISA). Out of 61 carnation
cultivars analyzed 41 (67%) were found positive. The virus positivity
was verified by polymerase chain reaction (PCR) and nucleotide
sequencing. The amplified 1349 bp fragment was by about 98% and 96%
identical with respect to coat protein (CP) and enzymatic polyprotein
genes, respectively, as compared to the sequences available in the
database. In terms of amino acid sequence similarity, the homology
values were 99% and 97%, respectively. Comparison with other
caulimoviruses revealed that CERV is most closely related to the
Cauliflower mosaic virus (CaMV). High genetic stability of CERV may be
attributed to the fact that it has evolved from the same initial
sequence in an original host. Because of global market of cut flowers
and vegetative propagation it has been dispersed around the world.
Key words: carnation; Carnation etched ring virus; coat
protein; enzymatic polyprotein; Indian isolate; nucleotide sequencing
Acta virologica 47: 105 – 111, 2003
R.M. de Campos, D.F. Ferreira, V.F. da Veiga, M.A. Rebello, M.C.S. Rebello*
Department of Virology, Prof. Paulo de Góes Institute of Microbiology, Federal University of Rio de Janeiro, Rio de Janeiro, CEP 21941-590, RJ, Brazil
Received March 3, 2003 accepted May 29, 2003
Summary. – The effect of
a cationic ionophore, monensin, on the replication of Mayaro virus
in monkey kidney TC7 and Aedes albopictus cells has been studied.
Treatment of these cells with 1 µmol/l monensin during infection
did not affect the virus protein synthesis but inhibited severely the
virus replication. Electron microscopy of the cells infected with Mayaro
virus and treated with monensin revealed that the morphogenesis of
Mayaro virus was impaired in TC7 but not in A. albopictus cells.
Key words: Mayaro virus; monensin; TC7 cells; Aedes
albopictus cells; protein synthesis; virus morphogenesis
Acta virologica 47: 113 – 119, 2003
O. Beran1*, M. Holub1, J. Špála1, J. Kalanin2, M. Staňková1
1 3rd Department of Infectious and
Tropical Diseases, First Faculty of Medicine, Charles University,
Budínova 2, Prague 8, CZ-180 81, Czech Republic;
2 Department of Immunology, Institute of Clinical and
Experimental Medicine; Prague, Czech Republic
Received February 14, 2003; accepted March 13, 2003
Summary. – The aim of this study
was to assess whether the density of CD38 antigen expression on CD8+ T
cells can be used as a marker of activation of the immune system in
Human immunodeficiency virus 1 (HIV-1)-positive patients treated with
highly active antiretroviral therapy (HAART). T cell subsets, expression
of CD38 antigen on CD8+ T cells, HIV-1 viral load and stage of the
disease were analyzed at baseline and after 12 months of HAART in 24
HIV-1-infected patients. Our data showed that the use of HAART is
effective in reducing plasma viral load and in achieving a stable
CD4+ count and percentage of CD8+/CD38+ cells. The percentages of
CD8+/CD38+ cells in HIV-1-infected patients at baseline and after 12
months of HAART were significantly higher than those of controls.
Analysis of the density of CD38 expression revealed that it was due to
CD8+/CD38+ subsets with low and medium density of antigen expression.
Absolute number of CD4+ T cells correlated negatively with the
percentage of CD8+/CD38+ cells at baseline of the study. Persistent
up-regulation of the CD38 expression on CD8+ T cells and its correlation
with the decreased CD4+ count despite the reduction of plasma viral load
may reflect residual replication of HIV-1 in reservoirs. Thus, this
immunological parameter can serve as a biological marker of HIV-1
infection and might have utility in clinical management of
HIV-1-infected persons.
Key words: HIV-1; CD38; CD8+; HAART; flow cytometry
Acta virologica 47: 121 – 124, 2003
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