[Use of recombinant protein E2 of the classical swine fever virus for immunization of animals]. / Primenenie rekombinantnogo belka E2 virusa klassicheskoi chumy svinei dlia immunizatsii zhivotnykh.

Recombinant major surface glycoprotein E2 from virulent Shimen strain of classical swine fever virus (CSFV) has been tested for immunogenicity in animal immunization experiments. Immunization of 3-month-old piglets with 200 micrograms of recombinant protein protected the animals from lethal challenge with virulent CSFV strain. CSFV-specific antibody detection test based on competitive ELISA has been developed using the recombinant E2 protein. The test can evaluate specific antibody levels after subunit vaccination with recombinant E2 after immunization with live vaccine based on attenuated CSFV strain.

[Synthesis and immunochemical properties of the recombinant major surface glycoprotein E2 of the classical swine fever virus]. / Sintez i immunokhimicheskie svoistva rekombinantnogo glavnogo poverkhnostnogo glikoproteina E2 virusa klassicheskoi chumy svinei.

Recombinant E2 protein from vaccine strain of classical swine fever virus (CSFV) and from SCFV virulent strain Shimen was synthesized in SF-21 and High-Five cell culture with baculovirus as the expressing vector. For secretion, hydrophobic C-terminal transmembrane domain was removed and N-terminal signal polypeptide of 38 amino acids was added. Maximum accumulation of recombinant products in SF-21 cells was observed after 48 h and in medium 96 h after infection with recombinant baculovirus. In High-Five cells and in culture medium the maximum accumulation of E2 was observed after 96 h. The level of E2 expression is 5-10 micrograms/106 cells. The products of expression were purified by affinity chromatography and their specificity confirmed in immunochemical tests with a series of reference monoclonal antibodies. The product can be used for detecting antibodies to SCFV by competitive enzyme immunoassay.

[Plasmapheresis in the combined therapy of progressive forms of glomerulonephritis]. / Plazmaferez v kompleksnoi terapii progressiruiushchikh form glomerulonefrita.

29 patients aged 6-16 with glomerulonephritis lasting 4-5 years received multimodality treatment with plasmapheresis as a component. The majority of the patients suffered from primary glomerulonephritis in mesangio- or membrano-proliferative morphological variants. Previous long-term conventional therapy (prednisolone, cytostatics, anticoagulants and antiaggregation drugs) failed. The test course comprised 1-3 plasmapheresis sessions (centrifuge method on [symbol: see text] apparatus), cyclophosphamide or maintenance methyl-prednisolone pulse therapy, heparin and curantil. One-third of the patients achieved remission lasting from 5 months to 3 years, in the other one-third the improvement was as short as 2-4 weeks, and the last one-third appeared non-responders. Improvement of clinical indices occurred in parallel with trends to reduction in the levels of CIC, IgG, B-lymphocytes, T-helpers, inhibition of lymphocyte succinate dehydrogenase activity, better phagocytosis. No complications which may prohibit plasmapheresis use in glomerulonephritis were observed. Adjuvant plasmapheresis use in glomerulonephritis treatment needs further studies.

[In vitro immunization of human lymphocytes with Epstein-Barr virus capsid antigen]. / Immunizatsiia in vitro limfotsitov cheloveka kapsidnymi antigenami virusa Epshteina-Barr.

Conditions for in vitro immunization of human lymphocytes from adult peripheral blood, tonsils and cord blood with Epstein-Barr Virus (EBV) capsid antigens have been studied. Pokeweed mitogen and B cell growth factor from Namalva cell line were shown to induce a significant production of specific antibodies by human lymphocytes stimulated with EBV. This effect made it possible to generate primary immune response in vitro using lymphocytes from EBV seronegative donors.

[Construction of a complete "library" of mutants at region -35 of a model prokaryotic promotor]. / Poluchenie polnoi "biblioteki" mutantov oblasti -35 model'nogo prokaryioticheskogo promotora.

A novel approach to study the variability of consensus sites of regulatory regions of DNA is proposed. The overall strategy includes chemical synthesis of representative series of all possible DNA structures under investigation, cloning and screening according to their function. The chemical-enzymatic synthesis of a complete library of 40-bp DNA duplexes, corresponding to the model prokaryotic promoter and differing in 6-membered segments at -35 region, is described.

A new class of antivirals: antisense oligonucleotides combined with a hydrophobic substituent effectively inhibit influenza virus reproduction and synthesis of virus-specific proteins in MDCK cells.

To enhance the penetration of oligonucleotide ('oligo') into cells, the oligo was combined with the hydrophobic undecyl residue. Using the 'DNA-synthesator', we synthesized oligo, complementary to the loop-forming site of the RNA, encoding polymerase 3 of the influenza virus (type A), and combined it with the undecyl residue added to the 5' terminal phosphate group. It was found that the modified oligo effectively suppresses the influenza A/PR8/34 (H1N1) virus reproduction and inhibits the synthesis of virus-specific proteins in MDCK cells. Under the same conditions, the non-modified antisense oligo and modified nonsense oligo did not affect the virus development.