Electronic Library of Scientific Literature - © Academic Electronic Press

Acta virologica

Volume 48 / 2004 / number 2

Here you can download FULL version of Acta Virologica 02/2004 in PDF format (parts are ordered by name of author): 
 Part 1 (Barde.pdf – 121 kB),  Part 2 (Hasmah.pdf – 143 kB),  Part 3 (Hosseini.pdf – 136 kB), Part 4 (Jirathitikal.pdf – 127 kB), Part 5 (Mosko.pdf – 393 kB), Part 6 (Pappova.pdf – 145 kB), Part 7 (Sierra.pdf – 161 kB), Part 8 (Stanekova.pdf – 93 kB), Part 9 (Subramanian.pdf – 150 kB), Part 10 (Zhao.pdf – 393 kB)
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Articles

Book Review

EFFECT OF AN ORAL THERAPEUTIC HIV-1 VACCINE ON AIDS PATIENTS WITH CD4 COUNT ABOVE 250 CELLS/MM3

V. JIRATHITIKAL1, A.S. BOURINBAIAR2*

1Immunitor Corporation, Ltd., 71 Moo 5, Bangpakong Industrial Park, Takarm, Chachoengsao 24130, Thailand;
2Immunitor USA Inc., College Park, MD 20740, USA

Received April 14, 2003; accepted March 16, 2004

Summary. – A simple delivery route, e.g. oral, would greatly facilitate the acceptance of an AIDS vaccine by a large target population. We developed a vaccine that took advantage of inherent properties of mucosal immunity in response to oral antigen challenge, namely the V-1 Immunitor vaccine (V1), which is a polyvalent oral Human immunodeficiency virus 1 (HIV-1) vaccine. The vaccine, currently manufactured in Thailand, contains pooled, inactivated viral antigens. In order to compare this vaccine with HIV-1 therapeutic vaccines reported earlier we analyzed retrospectively 13 HIV-1-positive patients that had the baseline CD4 T-cell counts greater than 250 cells/mm3 (range 270–605 cells/mm3). The patients self-administered one 850 mg vaccine tablet at breakfast and dinnertime for an average of 32 weeks (median 26 weeks). The treatment was well tolerated without any toxic effect. Twelve of thirteen patients (92%) and 9 of 13 patients (69%) experienced an elevation in CD4 and CD8 cells. The mean increase in absolute CD4 and CD8 cell counts across this group was 98 (22%; P = 0.02) and 324 (26%; P = 0.05) cells/mm3, respectively. Viral plasma load was measured by PCR in six patients. The observed viremia reduction was within 1 log unit. Subjective parameters, i.e., appetite, energy, and sense of well-being were reported by patients as being markedly improved, reflected in a mean body weight gain of 2.75 kg (P = 0.0008). Oral administration of HIV-1 immunogens provides compelling clinical response, especially when patients are treated earlier.
Key words: antiviral therapy; cellular immunity; clinical trials; gut, immune-based therapy; therapeutic vaccine

Acta virologica 48: 73 – 78, 2004

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MOLECULAR CHARACTERIZATION OF AN INFECTIOUS BURSAL DISEASE VIRUS ISOLATE FROM IRAN

S.D. HOSSEINI1,2, A.R. OMAR2*, I. AINI2

1Research Center of Animal Science and Natural Resources, Arak, Iran;
2Faculty of Veterinary Medicine, Universiti Putra Malaysia, 43400 UPM, Serdang, Selangor, Malaysia

Received November 21, 2003; accepted April 19, 2004

Acta virologica 48: 79 – 83, 2004

Summary. – The segment A of an Infectious bursal disease virus (IBDV) isolate from Iran was amplified by reverse transcription–polymerase chain reaction (RT-PCR), sequenced and compared with published sequences of 26 IBDV isolates from other parts of the world. The Iranian isolate showed 8 unique amino acid differences. In addition, 9 common amino acid differences, namely 3 in VP2, (222 Ala, 256 lIe and 294 lIe), 3 in VP4 (685 Asn/Ser, 715 Ser and 751 Asp), 2 in VP3 (990 Val and 1005 Ala), and 1 in VP5 (49 Arg) were found. Phylogenetic analysis indicated that the Iranian isolate is closely related to highly virulent (hv) IBDV isolates from Asian countries. Nevertheless, it may share a common origin with hv isolates from other parts of the world.
Key words: Infectious bursal disease virus; segment A; RT-PCR; nucleotide sequence; phylogenetic analysis

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GENETIC DIVERSITY OF CHICKEN ANEMIA VIRUS FOLLOWING CELL CULTURE PASSAGING IN MSB-1 CELLS

M.S. HASMAH, A.R. OMAR*, K.F. WAN, M. HAIR-BEJO, I. AINI

Faculty of Veterinary Medicine, Universiti Putra Malaysia, 43400 UPM, Serdang, Selangor, Malaysia

Received October 16, 2003; accepted April 29, 2004

Summary. – It has been shown that a chicken anemia virus (CAV) isolates which had undergone 60 passages in MSB-1 cells (SMSC-1/P60, 3-1/P60) acquired 33–66 nucleotide substitutions at the coding region resulting in 13–16 amino acid changes as compared to the CAV isolates passaged only 5 times in MSB-1 cells (SMSC-1 and 3-1) (Chowdhury et al., Arch. Virol. 148, 2437–2448, 2003). In this study we found that a low CAV (BL-5) and a high CAV passage (BL-5/P90) differed by only 15 nucleotide substitutions resulting in 11 amino acid changes. Phylogenetic analysis based on VP1 also revealed that both isolates were close to each other but not to other CAV isolates from Malaysia, namely SMSC-1 and 3-1.
Key words: Chicken anemia virus; virus passaging; nucleotide substitutions; amino acid substitutions; coding regions, nucleotide sequence; phylogenetic analysis

Acta virologica 48: 85 – 89, 2004

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PATHOGENETICAL CHARACTERIZATION OF ISOLATE MHV-60 OF MOUSE HERPESVIRUS STRAIN 68

M. PAPPOVÁ1, M. STANČEKOVÁ1, I. SPIŠŠÁKOVÁ2, V. ĎURMANOVÁ2, J. MISTRÍKOVÁ1,2*

1Department of Microbiology and Virology, Faculty of Natural Sciences, Comenius University, Mlynská dolina B2, 842 15 Bratislava, Slovak Republic;
2Institute of Virology, Slovak Academy of Sciences, Dúbravská cesta 9, 842 05 Bratislava, Slovak Republic

Received January 28, 2004; accepted May 4, 2004

Summary. – Infection of mice with mouse herpesvirus strain 68 (MHV-68) is an excellent small animal model of gammaherpesvirus pathogenesis in a natural host. We carried out comparative studies on MHV-60, another isolate of MHV-68. The acute infection of BALB/c mice inoculated intranasally (i.n.) with MHV-60 as well as its impact on tumor development were investigated. During the acute phase of infection the lungs were the main tissues infected. Our results show that MHV-60 has similar pathological features like other 4 isolates so far examined, namely MHV-72, MHV-78, MHV-Šumava inclusive of MHV-68. Nevertheless, MHV-60 differed from other isolates in following features: (i) the acute phase of infection was established very soon and lasted 10 days post infection (p.i.) in contrast to 14–28 days p.i. in the abovementioned isolates with a peak on days 3–5 p.i. The virus could also be recovered from the spleen, thymus and kidneys but not in other investigated organs. A lymphoproliferative response was associated with splenomegaly. At this time an increase in the number of leukocytes and appearance of atypical leukocytes in peripheral blood were observed. (ii) the infection was localized in the lungs and spleen, while in other isolates it was detected in a much broader scale of organs, and (iii) the acute phase of infection was accompanied by a massive splenomegaly, which was characteristic for the chronic phase of infection. Despite the fact that after clearance of the acute infection the virus was hardly detected, the tumor formation was later observed in 22% of infected mice as compared to 5% in control non-infected mice.
Key words: MHV-60; MHV-68; pathogenesis; tumors; lymphoma

Acta virologica 48: 91 – 96, 2004

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EXPRESSION OF HERPES SIMPLEX VIRUS 1 GLYCOPROTEIN D IN PROKARYOTIC AND EUKARYOTIC CELLS

T. MOŠKO1, J. KOŠOVSKÝ2, I. REŽUCHOVÁ2, V. ĎURMANOVÁ2, M. KÚDELOVÁ2, J. RAJČÁNI2*

1Institute of Microbiology and Immunology, Jessenius Faculty of Medicine, Comenius University, Martin, Slovak Republic;
2Institute of Virology, Slovak Academy of Sciences, Dúbravská cesta 9, 845 05 Bratislava, Slovak Republic

Received January 28, 2004; accepted May 13, 2004

Summary. – Recombinant plasmids encoding either the full-length glycoprotein D (FLgD) or truncated gDs were constructed. The recombinant plasmids were expressed in Escherichia coli and BHK-21 cells. The strongest expression was obtained with the recombinant plasmid encoding a truncated gD which corresponded to the gD ectodomain. The cells transformed with this plasmid showed good exponential growth ensuring satisfactory yields of the expressed polypeptide in the form of the fusion protein. The fusion protein was biotinylated and efficiently purified. The shortest truncated gD, which contained the main continuous antigenic locus VII binding neutralization antibody and additional continuous antibody binding epitopes, still reacted with specific antibody as proven by immunoblot analysis. In addition, a shuttle vector for expression of FLgD in mammalian cells was constructed. This vector-transfected BHK-21 cells expressed gD for 40 days during 9 consecutive passages. The expression of gD began on day 2 and culminated at day 9 post transfection (p.t.).
Key words: BHK-21 cells; Escherichia coli; expression; fusion protein; glycoprotein D; Herpes simplex virus 1; recombinant plasmid; shuttle vector

Acta virologica 48: 97 – 107, 2004

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THE ROLE OF HEMAGGLUTININS IN THE MIDGUT EXTRACTS OF TWO LINES OF AEDES AEGYPTI IN THEIR SUSCEPTIBILITY TO DENGUE-2 VIRUS

P.V. BARDE1, M.I. KHAN2, M.D. GOKHALE1, A.C. MISHRA1, D.T. MOURYA1*

1National Institute of Virology, 20-A, Dr. Ambedkar Road, Pune, India;
2Division of Biochemistry, National Chemical Laboratory, Pashan Road, Pune 411 008, India

Received February 25, 2003; accepted May 25, 2004

Summary. – Hemagglutinin activity (HA) was studied in the midgut extracts from highly (h) and lowly (l) susceptible strains of Aedes aegypti mosquitoes to Dengue-2 virus (DEN-2). HA in the midgut extracts from these two isofemale strains of mosquitoes was high in (l) as compared to (h) mosquitoes. HA was found to be higher with chicken red blood cells (RBCs) than with rabbit and human RBCs of O group. Larval midgut extracts showed higher activity than those from adult female mosquitoes. Exposure of midgut extracts to 100oC for 10 mins destroyed the activity. The activity was observed between pH 6 and pH 10. HA in midgut extracts was also studied using twenty different carbohydrates; five of them showed an inhibition of HA. The inhibitory carbohydrates, when incorporated into DEN-2-infected bloodmeal, showed a reduction in the susceptibility of mosquitoes to the virus as compared to the control ones fed on the virus alone. Similarly, when these carbohydrates were incorporated in the DEN-2-infected inoculum, the inoculated mosquitoes showed a reduction in the susceptibility to the virus. HA in the virus-infected midgut extracts was higher than that in the uninfected controls. These results suggest that the presence of HA in the midgut may be one of the factors that affect the susceptibility of Ae. aegypti mosquitoes to DEN-2.
Key words: Aedes aegypti; carbohydrates; dengue; Dengue-2 virus; hemagglutinin; mosquito

Acta virologica 48: 109 – 113, 2004

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GENETIC STABILITY OF THE ATTACHMENT GLYCOPROTEIN OF HUMAN RESPIRATORY SYNCYTIAL VIRUSES DURING SERIAL PASSAGES IN CELL CULTURES

M. DE SIERRA, J. ARBIZA*

Sección Virología, Facultad de Ciencias, Universidad de la República, Igua 4225 (11400), Montevideo, Uruguay

Received November 18, 2003; accepted May 25, 2004

Summary. – Thirteen isolates of human respiratory syncytial viruses (HRSV) of groups A and B were isolated in HEp-2 cells from nasopharyngeal aspirates (NPA) from the children with acute respiratory infections. Three isolates of HRSV of group A were propagated in HEp-2 cells in 20 serial passages. Nucleotide sequences of the products obtained by RT-PCR from the glycoprotein (G) hypervariable region of the original virus isolates in NPA and those after one or several passages were compared. All the isolates analyzed showed no changes during passaging in HEp-2 cells.
Key words: human respiratory syncytial viruses; genetic stability; attachment glycoprotein

Acta virologica 48: 115 – 121, 2004

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INTERACTION BETWEEN GENOMES OF INFECTIOUS BRONCHITIS AND NEWCASTLE DISEASE VIRUSES STUDIED BY REVERSE TRANSCRIPTION–POLYMERASE CHAIN REACTION

B.M. SUBRAMANIAN, G.D. RAJ*, K. KUMANAN, K. NACHIMUTHU, A.M. NAINAR

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

Received October 16, 2004; accepted June 11, 2004

Summary. – Reverse transcription–polymerase chain reaction (RT-PCR) with specific primers for the S1 gene of IBV and for the fusion protein cleavage site of NDV was used for detection of Infectious bronchitis virus (IBV, the family Coronaviridae) and Newcastle disease virus (NDV) genomes. The sensitivity of IBV and NDV RT-PCR was 103.7 and 103.0 EID50, respectively. Although a multiplex RT-PCR could detect and differentiate NDV and IBV genomes present in the same sample, there was a slight inhibition of the IBV PCR if a high amount of NDV genome was present in the sample. To overcome this problem a separate PCR for each virus was used to assess the interaction between vaccine IBV and NDV either inoculated singly or together into chickens. In the group vaccinated with the Newcastle disease (ND) vaccine alone, the viral genome was detected on days 2, 4 and 7 post_vaccination (p.v.), while in the chickens given the infectious bronchitis (IB) vaccine alone, the viral genome was detected only on day 4 p.v. In the group inoculated with both vaccine viruses there was a 103-fold reduction in the cDNA dilution factor on day 4 p.v. for both IBV and NDV genomes. This demonstrated clearly that when both these vaccines are administered there is a transient reduction in the replication of both viruses, probably due to their competition for the same target epithelial cells in the respiratory tract.
Key words: RT-PCR; Infectious bronchitis virus; Newcastle disease virus; virus interaction; cDNA dilution factor

Acta virologica 48: 123 – 129, 2004

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