Finnish-Russian programme for the control of rabies (2005-2010).

The Finnish-Russian collaboration on rabies control began in 2000. This data summarizes the results of the scientific part of the programme, including rabies monitoring in Russia and the molecular epidemiological studies with field viruses.

[Production of monoclonal antibodies to the prion protein and their characterization].

Antibodies to the prion protein (PrP), particularly, monoclonal antibodies, are necessary tools in the diagnostics and study of prion diseases and potential means of their immunotherapy. For the production of monoclonal antibodies, BALB/c mice were immunized by a recombinant bovine PrP. Three stable hybridomas producing antibodies of IgM class were prepared. The antibodies were bound to PrP in a solid-phase enzyme immunoassay and immunoblotting. The epitope mapping accomplished with the use of synthetic peptides showed that an epitope located in region 25-36 of PrP corresponds to one antibody, and epitopes located in region 222-229, to the other two. The antibodies to fragment 222-229 purified by affinity chromatography recognized with a high specificity conglomerates of a pathogenic prion in the brain tissue of cows suffering from spongiform encephalopathy. Thus, in nontransgenic mice, PrP-specific monoclonal antibodies were produced, useful in studies and diagnostics of prion diseases.

[Primary structure of the A22 RNA polymerase gene of the foot and mouth disease virus]. / Pervichnaia struktura gena RNK-polimerazy virusa iashchura A22.

Complete nucleotide sequence of gene RNA polymerase for the foot-and-mouth disease virus subtype A22 has been determined.

[Synthesis and immunogenic properties of peptides--fragments of the immunodominant regions of the VP1 protein of the Asia-1 type of foot- and-mouth disease virus]. / Sintez i immunogennye svoistva peptidov--fragmentov immunodominantnykh raionov belka VP1 virusa iashchura tipa Aziia-1.

Potential immunodominant epitopes were predicted on the basis of a theoretical analysis of the antigenic structure of the VP1 protein of the type Asia-1 foot-and-mouth disease virus. Peptides corresponding to the 140-153, 136-153, 132-153, 143-157, 137-157, and 193-208 fragments of the VP1 protein sequence were synthesized by the solid phase method, and the immunogenic properties of the peptides were studied on guinea pigs. The shortest peptide exhibiting the protective effect was found to correspond to the, 140-153 fragment of the VP1 sequence. The Plm-(Gly)3-(140-153)-(Gly)2-Lys(Plm)-Leu and [Ac-(140-153)-(Gly)3]8-(Lys)7-Gly synthetic constructions in combination with adjuvants provided up to 80% protection of immunized animals against infection with the foot-and-mouth disease virus.

[Protection from foot-and-mouth disease virus in naturally-susceptible animals by a linear polymer of a synthetic peptide]. / Zashchita ot iashchura estestvenno-vospriimchivykh zhivotnykh lineinym polimerom sinteticheskogo peptida.

Linear polymer of a peptide corresponding to the fragment 142-155 of the foot-and-mouth disease virus A22(550) protein (VP1) was synthesized. Whereas the monomeric peptide was only slightly immunogenic, the polymer induced virus-neutralizing antibodies in rabbits and protected 100% guinea pigs. Sheep vaccinated once and cattle vaccinated twice were stable against infection with the homologous virulent foot-and-mouth disease virus.

[Protection of naturally susceptible animals against foot-and-mouth disease with a peptide, synthesized on a lysine matrix]. / Zashchita estestvenno-vospriimchivykh zhivotnykh ot iashchura peptidom, sintezirovannym na lizinovoi matritse.

A peptide VP1-(142-158)-MAP (Multiple antigen peptide system) consisting of two parts: a lysine matrix made up of three levels of lysine residues coupled with each other and amino acid sequence 142-158 of VP1 of FMD virus strain A(22)550--has been synthesized. Guinea-pigs inoculated with 20 mkg of the peptide incorporated with Freund's complete adjuvant were protected against challenge with 500 ID50 of homologous FMD virus. Sheep were immunized with a single inoculation of the peptide in a dose of 1.0 mg. Cattle inoculated twice with 1.5 mg of the peptide with incomplete adjuvant on the basis of synthetic oil developed high virus-specific antibody titres both after the first (5.3-7.6 log2 ND50/0.1 ml) and the second inoculation (10.2-11.0 log2 ND50/0.1 ml). The peptide-immunized animals were resistant to challenge with homologous virulent virus in a dose of 10(4) ID50. The immunogenic and protective capacities of the peptide VP1-(142-158)-MAP were shown to be greater as compared with those of its linear analogue-peptide VP1-(141-160).

[Induction of immune response by synthetic fragments of the bovine prion protein and their analogues in mice of various lines]. / Induktsiia immunogo otveta sinteticheskimi fragmentami prionnogo belka i ikh analogami u myshei raznykh linii.

The antibodies to the bovine prion protein were produced by immunizing mice of three lines with five synthetic fragments of the protein and their six analogues. The analogues contained the amino acid substitutions that, according to theoretical calculation, should lead to an increase in the immunogenic activity of peptides. All the peptides, except for one, induced the formation of antibodies. All the sera containing the antipeptide antibodies were tested by an immunohistochemical method. The sera that were effectively bound to the brain preparations from the bovine with spongiform encephalopathy were identified; it was shown that they do not interact with the preparations of normal brain. Therefore, it was shown that the immunization of mice with the synthetic fragments of a prion protein helps obtain specific antibodies suitable for the study and diagnostics of prion diseases.

[Nucleotide sequence of the RNA polymerase gene attenuated by a variant of foot-and-mouth disease virus and its comparison with the virulence of related variants of subtype A22]. / Nukleotidnaia posledovatel'nost' gena RNK-polimerazy attenuirovannogo varianta virusa iashchura i ee sravnenie s virulentnymi rodstvennymi variantami podtipa A22.

The nucleotide sequence of RNA-polymerase gene and 3'-terminal untranslated genome region of attenuated foot-and-mouth disease virus (FMDV) strain A(22)645 has been determined. RNA-polymerase gene and predicted amino acid sequences of attenuated FMDV strain A(22)645 were compared with those of the original virulent FMDV strain A(22)550. The examined genome region of strain A(22)645 differed from that of strain A(22)550 by 22 nucleotides and 3 amino acids. Three mutations occurred within nucleotide residues 883 to 1026, which encode an extremely conserved amino acid domain corresponding to RNA polymerization and nucleoside triphosphate binding functions. Mutation in 920n caused substitution of conserved amino acid Asn for Ser in the core motif for nucleotide binding Gxxx-TxxxN(S/T). It is possible that change in functional motif caused the alteration of phenotypic virus and loss of pathogenicity for cattle. This may be finally confirmed only after comparative analysis of complete genomes of the examined virus strains.

[The use of synthetic peptides for detection of antibodies against foot-and-mouth disease virus in the blood from convalescent animals]. / Ispol'zovanie sinteticheskikh peptidov dlia vyiavleniia antitel k virusu iashchura v syvorotkakh krovi perebolevshikh zhivotnykh.

Peptides were synthesized, which, according to theoretical analysis of the antigenic structure of protein VP1 of foot-and-mouth disease (FMD) virus types A, 0, and Asia 1, corresponded to potential immunodominant protein sites. Activities of the peptides were studied by solid-phase indirect radioimmunoassay on polyethylene film with purified immunoglobulins against intact FMD virus. Virtually no cross reactions were observed. Blood sera of cattle convalescent after FMD were tested with the FMD virus and peptides containing VP1 fragments 141-160 (A22 No. 550), 140-160 (O1 No. 194), and 140-153 (Asia 1 No. 48). The specificity of interactions between the sera and peptides and the virus was uniform, this permitting the identification of the virus type which caused the disease.

[Cloning fragments of the RNA polymerase gene of an attenuated variant of the foot-and-mouth disease virus A22]. / Klonirovanie fragmentov gena RNK-polimerazy attenuirovannogo varianta virusa iashchura A22.

The cDNA fragments complementary to RNA-polymerase gene and 3'-untranslated genome region of attenuated foot-and-mouth disease virus strain A(22)645 have been synthesized and cloned into a plasmid vector pUC19 in E. coli JM109. The cloned cDNA fragments were characterized as to their size, orientation towards the plasmid, and localization in the virus genome. Restriction maps for complete gene and two cDNA clones were constructed.

Monoclonal antibody characterization of rabies virus isolates from Russia, Finland and Estonia.

Five different rabies virus variants were identified among rabies virus-positive samples from Russia, Finland and Estonia, using a panel of five anti-nucleocapsid monoclonal antibodies. Two rabies virus isolates showed a different reaction pattern, suggesting the presence of a new antigenic variant. The results were compared with the data obtained by other research groups.

[Primary structure of the 3'-terminal region of the RNA-polymerase gene in foot-and-mouth virus A22]. / Pervichnaia struktura 3'-kontsevoi oblasti gena RNK-polimerazy virusa iashchura A22.

This study was undertaken for the purpose of determining the primary structure of the 3' end gene of RNA polymerase of foot-and-mouth disease virus A22 550. Reported are isolation and purification of the virus, isolation of RNA, synthesis of cDNA, experience obtained from cloning as well as analysis of hybridisation and isolation of plasmid DNA. Nucleotide sequences, characterised by specific clones, were tested for potential needle structures. Also described are homology comparisons among FMD virus types A12, A10, O1, and C1.