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Volume 46 / 2002 / number 1
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Q. ZHANG, Y.Q. YANG, Z.Y. ZHANG, L.LI, W.Y. YAN, W.J. JIANG, A.G. XIN, C.X. LEI, Z.X. ZHENG
State Key Laboratory of Genetic Engineering,
School of Life Science, Fudan University, Shanghai 200433, P.R. China;
Yunnan Tropical and Subtropical Animal Virus Diseases Laboratory,
Kunming, Yunnan, P.R. China
Summary. – In this study, the sequences of capsid protein VP1 regions of YNAs1.1 and YNAs1.2 isolates of foot-and-mouth disease virus (FMDV) were analyzed and a peptide containing amino acids (aa) 133–158 of VP1 and aa 20~34 of VP4 of FMDV type Asia 1 was assumed to contain B and T cell epitopes, because it is hypervariable and includes a cell attachment site RGD located in the G-H loop. The DNA fragments encoding aa 133–158 of VP1 and aa 20~34 of VP4 of FMDV type Asia 1 were chemically synthesized and ligated into a tandem repeat of aa 133–158–20~34–133–158. In order to enhance its immunogenicity, the tandem repeat was inserted downstream of the beta-galactosidase gene in the expression vector pWR590. This insertion yielded a recombinant expression vector pAS1 encoding the fusion protein. The latter reacted with sera from FMDV type Asia 1-infected animals in vitro and elicited high levels of neutralizing antibodies in guinea pigs. The T cell proliferation in immunized animals increased following stimulation with the fusion protein. It is reported for the first time that a recombinant fusion protein vaccine was produced using B and T cell epitopes of FMDV type Asia 1 and that this fusion protein was immunogenic. The fusion protein reported here can serve as a candidate of fusion epitopes for design of a vaccine against FMDV type Asia 1.
Key words: FMD;
FMDV type Asia 1; VP1; epitope; recombinant protein; immunogenicity;
guinea pig
Acta virologica 46: 1 – 9, 2002
H.W. ZHANG, Z.C. QU, X.Y. XU, X.N. ZHANG, F.W. BAI, Y.Z. WAN, M.H. SHAO, M.M. YE, D.L. SHEN
Institute of Genetics, School of Life Sciences, Fudan University, Shanghai 200433, P.R. China
Summary. – Phages with high affinity to the P20 protein of rice stripe virus (RSV) were enriched from phage-displayed random 12-mer peptide library after three rounds of phage display screening. Nine different peptides from the enriched library were selected by enzyme-linked immunosorbent assay (ELISA). The P20 protein from raw extracts of rice leaves infected with RSV could be detected by those 9 peptides displayed on the phage, which suggested that a peptide could be an effective tool for diagnosis of RSV in rice and planthopper. Circular dichroism (CD) spectra of P20 fusion proteins with the binding phages and non-binding phages showed that the conformation of P20 protein was changed after binding to each of the 9 selected 12-mer peptides, which suggested that these peptides might disrupt the function of the P20 protein. Thereafter, those peptides might be used to develop plant resistance and disrupt virus transmission. Three of the 12-mer peptide genes were fused with the glutathione-S-transferase (GST) gene in the vector pGEX 3X. The fusion proteins were obtained from an Escherichia coli expression system and purified. The fusion proteins might have a potential to develop a plant peptide-based resistance to its pathogens and virus diagnosis. It also provided a tool (i) to confirm the inhibition of the function of P20 protein by the fusion peptides in vivo, and (ii) to detect the function of P20 protein and the interaction between the virus and its vector.
Key words: peptide;
phage display; P20 protein; protein conformation; rice stripe virus
Acta virologica 46: 11 – 17, 2002
M. MRMUSOVÁ, M. HORVÁTHOVÁ, M. KLOBUŠICKÁ, J. MISTRÍKOVÁ
Department of Microbiology and Virology, Faculty
of Natural Sciences, Comenius University, Mlynská dolina B2, 842 15
Bratislava, Slovak Republic;
Institute of Virology, Slovak Academy of Sciences, Dúbravská cesta 9,
842 45 Bratislava, Slovak Republic;
Department of Clinical Immunology, Institute of Preventive and Clinical
Medicine, Bratislava, Slovak Republic;
Institute of Experimental Oncology, Slovak Academy of Sciences,
Bratislava, Slovak Republic
Summary. – We characterized leukocytes in peripheral blood of BALB/c mice infected with mouse herpesvirus isolate 72 (MHV-72) representing an isolate of mouse herpesvirus strain 68 (MHV-68, species Murid herpesvirus 4, genus Rhadinovirus, subfamily Gammaherpesvirinae, family Herpesviridae) (van Regenmortel et al., 2000). In acute infection (up to day 30 post infection (p.i.)) the number of CD8+ T cells increased, reaching a maximum at day 11 p.i. This increase correlated with that of CD4+ T, activated CD19+ B and natural killer (NK) cells. At day 30 p.i. the numbers of CD4+, CD8+, CD14+ and CD19+ cells decreased to normal values. A similar increase in the number of these cells was observed at day 730 p. i. In the course of persistent infection (after day 30 p.i.) some of the mice developed a leukemia-like syndrome characterized by an increase in the number of leukocytes and appearance of atypical, blastic immature forms of leukocytes. The latter forms of leukocytes were characteristic by an increased amount of argyrophilic proteins. These results show further similarities between MHV-72 (another isolate of MHV-68) and EBV infections and justify the use of MHV-68 or MHV-72 as an appropriate mouse model for the study of EBV infection of humans.
Key words: argyrophilic
proteins; blood picture; immunophenotyping; mouse herpesvirus isolate 72
(MHV-72); mouse herpesvirus strain 68 (MHV-68); infectious
mononucleosis-like syndrome
Acta virologica 46: 19 – 24, 2002
J.Z. ZHANG, M.Y. FAN
Rickettsial laboratory, Institute of Epidemiology and Microbiology, Chinese Academy of Preventive Medicine, Beijing 102206, P.R. China
Summary. – Pulsed-field gel electrophoresis (PFGE) was used to determine the genome size and the restriction fragment length polymorphism (RFLP) of four new Chinese isolates of spotted fever group (SFG) rickettsiae. The genome size of the isolates Ha-91, BJ-90, 054 and 036 was 1253 kb, 1236 kb, 1272 kb, and 1272 kb, respectively. The isolates 054 and 036 had identical RFLP profiles. All the isolates differed in the properties under study from the so far known SFG rickettsiae. The unique RFLP profiles of the isolates supported our opinion that they are new strains of SFG rickettsiae or even new species of SFG rickettsiae.
Key words: rickettsiae;
spotted fever group; Chinese isolates; pulsed-field gel electrophoresis
Acta virologica 46: 25 – 30, 2002
E. MITROVÁ, G. BELAY
Institute of Preventive and Clinical Medicine, Limbová 14, 833 01 Bratislava, Slovak Republic
Summary. – Creutzfeldt-Jakob disease (CJD), the most important human prion disease, occurs in sporadic, iatrogenic and familial form. Except Slovakia and Israel, the recorded familial cases have never exceeded 10–15%. In the Slovak CJD group 95 out of 136 CJD cases (74.2%) carried a CJD-specific mutation in the prion protein gene (PRNP) at codon 200 (mutation E200K). All CJDE200K patients carried a heterozygous E200K mutation within the alelle with methionine at codon 129. No more than 53.7% were typical familial cases. The penetrance of the E200K mutation in 1975–2000 was 59.5%. The distribution of codon 129 polymorphism showed 78.6% of methionine-homozygous and 21.4% of methionine/valine-heterozygous patients. Genetic analysis performed on 278 CJD patient relatives demonstrated the E200K mutation in 97 (34.8%) of healthy relatives tested. The E200K mutation carriers were methionine-homozygous in 64% and methionine/valine-heterozygous in 36%. The relatives without the mutation showed a 54.9% methionine homozygosity, 10.4% valine homozygosity and 34.7% methionine/valine heterozygosity. Analysis of the E200K carriers provided evidence that the methionine homozygosity is a CJD risk factor, more efficient in CJD patients than in asymptomatic relatives. Th influence of both the E200K mutation and methionine homozygosity at codon 129 was evident in the duration of the clinical stage of CJD and in the immunoreactivity pattern of PrP resistant to proteases (PrPres). In the CJDE200K methionine-homozygous patients the mean duration of the disease was significantly shorter (3.7±2.0 months) than in the methionine/valine-heterozygous patients (7.84±7.3 months). Comparison of the PrPres positivity in the cerebellum of familial and sporadic CJD using specific polyclonal and monoclonal antibodies (MAbs) to PrP showed less conspicuous immune reaction in CJDE200K cases. Methionine-homozygous CJD patients were characteristic mainly by synaptic pattern of staining, while methionine/valine-heterozygous patients by PrPres granules and plaque-like structures. Most of numerous plaque-like PrPres deposits were found in sporadic valine/valine-homozygous cases. Potential professional risk was excluded in health facility workers. The percentage of professions related to farming was significantly higher in CJDE200K (48%) and sporadic CJD (44%) cases as compared to the employed population (9%).
Key words: Creutzfeldt-Jakob
disease; genetic form; E200K mutation; penetrance; potential
professional risk; Slovakia
Acta virologica 46: 31 – 39, 2002
J. MISTRÍKOVÁ, T. MOŠKO, M. MRMUSOVÁ
Department of Microbiology and Virology, Faculty of Natural Sciences, Comenius University, Mlynská dolina B2, 842 15 Bratislava, Slovak Republic
Summary. – BALB/c mice inoculated intranasally (i.n.) with the mouse herpesvirus isolate Šumava (MHV-Šumava) did not show apparent symptoms of illness. However, they showed an increase in the number of leukocytes and appearance of atypical leukocytes in peripheral blood. The infiltration of spleen with atypical cells resulted in splenomegaly. In the course of the infection the virus persisted in lungs, spleen, thymus, bone marrow, mammary glands, peritoneal macrophages and liver. We regard MHV-Šumava as one of the eight isolates of MHV-68 (a virus and species Murid herpesvirus 4, genus Rhadinovirus, subfamily Gammaherpesvirinae, family Herpesviridae (van Regenmortel et al., 2000)) so far obtained. MHV-Šumava differs from MHV-68, MHV-72 and MHV-76 in some virological and pathogenetical features.
Key words: mouse
herpesvirus; isolate Šumava; pathogenesis, lymphatic system
Acta virologica 46: 41 – 46, 2002
A.Y. HUI, F.K.L. CHAN, P.K.S. CHAN, J.S.L. TAM, J.J.Y. SUNG
Departments of Medicine and Therapeutics and Microbiology, Prince of Wales Hospital, Chinese University of Hong Kong, Shatin, Hong Kong
Key words: hepatitis
C virus; serology; whole-blood test
Acta virologica 46: 47 – 48, 2002
E. ŠPITALSKÁ, E. KOCIANOVÁ
Institute of Virology, Slovak Academy of Sciences, Dúbravska cesta 9, 842 45 Bratislava, Slovak Republic
Key words: Ehrlichia
phagocytophila; ticks; Slovakia
Acta virologica 46: 49 – 50, 2002
D.T. MOURYA
National Institute of Virology, 20-A, Dr. Ambedkar Road, Pune 411 001, India
Key words: anticoagulants;
Aedes aegypti; dengue virus; hemorrhagic fever; virus transmission;
viremia
Acta virologica 46: 51 – 53, 2002
H.J. Schurman, G. Feutren, J.F. Bach (Eds): Modern Immunosuppressives. The series: M.J. Parnham and J. Bruinwels (Eds): Milestones in Drug Therapy. Brikhäuser Verlag, Basel-Boston-Berlin, 2001, 218 pp.
Acta virologica 46: 55, 2002
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