[A comparison of two existing catalogues of the alleles of gliadin-coding loci in winter common wheat].

Two catalogs of alleles of gliadin-coding loci, controlling synthesis of a storage protein of wheat caryopsis, gliadin, were compared. One catalogue comprises the alleles detected according to the electrophoretic patterns in starch gels; the other, in polyacrylamide gels. Determination of the allelic state of gliadin-coding loci in 31 previously not studied cultivars of winter common wheat allowed us to construct a matching system for the alleles compiled in the two catalogs, which gives the possibility to compare the results of wheat cultivar analyses performed at different scientific institutions.

[Kama, a new virus (Flaviviridae, Flavivirus, Tiulenii antigenic group), isolated from Ixodes lividus ticks]. / Novyi virus kama (Flaviviridae, Flavivirus, antigennaia gruppa tiulenii), izolirovannyi iz kleshchei Ixodes lividus.

Three identical strains of a new virus Kama (Flaviviridae, Flavivirus, Tyuleny antigenic group) were isolated from Ixodes lividus Roch, obligate parasites of Riparia riparia L. The ticks were collected in June, 1990 in Tatarstan on the islands in the basin of the Kama river. The strains were examined under electron microscope and by serological tests (neutralization, complement fixation, hemagglutination inhibition, and indirect immunofluorescence). The virus is antigenically related but not identical to Tyuleny virus. Hence, the Tyuleny antigenic group at present includes viruses Tyuleny, Meaban, Gadgest Gally, Saumares Reef, and Kama. All these viruses are associated with an ecosystem including ixodide ticks, obligate parasites of colonial birds.

[Analysis of the stability of some properties of tick-borne encephalitis virus (strain 205) upon passaging in mice]. / Analiz stabil'nosti nekotorykh svoistv virusa kleshchevogo éntsefalita (shtamm 205) pri passirovanii na myshakh.

Comparative analysis of the characteristics of tick-borne encephalitis (TBE) strain 205 used for the production of vaccine against TBE and of its variants obtained by passages in mouse brain showed the stability of such properties as infective activity, neurovirulence, sensitivity to physical (heating to 50 C) and chemical (sodium deoxycholate treatment) factors. At the same time increased neurovirulence of variants 205/M10 and 205/M20, which undergone through 10 and 20 passages in white mouse brain, for low-sensitive Syrian hamsters was revealed. Use of a panel of 5 monoclonal antibodies to protein E and of 4 monoclonal antibodies to protein E and of 4 monoclonal antibodies to protein NS3 helped differentiate between not only strains 205 and Sofyin, but between variants of strain 205 as well.

[The detection of antibodies to the Venezuelan equine encephalomyelitis virus by immunoenzyme analysis using antigen purified in a water-polymer system]. / Vyiavlenie antitel k virusu venesuél'skogo éntsefalomielita loshadei metodom immunofermentnogo analiza s primeneniem ochishchennogo v vodno-polimernoi sisteme antigena.

Human and animal sera were tested for the presence of antibodies to Venezuelan equine encephalomyelitis (VEE) virus by direct enzyme immunoassay (EIA). For the test, the plates were sensitized with a VEE virus preparation purified in a two-phase system of water-soluble polymers. The proposed EIA variant was as specific as that with VEE antigen obtained by fractionation on sucrose cushion, but more sensitive. The high specificity of the assay allowed the antigen purified in the water-polymer system to be used for investigation of antigen relationships among viruses of the VEE complex.

[A monoclonal antibody study of the receptor area of the Venezuelan encephalitis virus]. / Izuchenie retseptornoi oblasti virusa venesuél'skogo éntsefalita loshadei pri pomoshchi monoklonal'nykh antitel.

The receptor region for virus-cell interaction in Venezuelan equine encephalomyelitis (VEE) and Eastern equine encephalomyelitis (EEE) viruses was studied using a panel of 17 monoclonal antibodies (MCA). They were able to block agglutination of goose erythrocytes. The dominant role of glycoprotein E2 in the formation of viral receptor for EEE and VEE viruses was demonstrated. Competitive radioimmunoassay identified three antigenic sites in this region. These sites were also responsible for virus neutralization. MCAs to these sites protected outbred mice against lethal infection. The presence of a highly conservative region in VEE (site E2-3) and EEE (site E2a) which produced cross-reacting antibodies blocking hemagglutination of Western equine encephalomyelitis, Semliki Forest, Sindbis, Getah, Aura, Chikungunya, and Pixuna viruses was established. A hypothesis is suggested concerning the existence of similar regions for the entire alphavirus genus, and the role of this region in virus-cell interaction.

[The demonstration of the Venezuelan equine encephalomyelitis virus by the method of indirect lanthanide immunofluorescent analysis]. / Indikatsiia virusa venesuél'skogo éntsefalomielita loshadei metodom nepriamogo lantanidnogo immunofliuorestsentnogo analiza.

A test system of indirect time-resolved fluoroimmunoassay (TR-FIA) was developed and tested on an alpha-virus, Venezuelan equine encephalomyelitis virus. The indirect TR-FIA test system was shown to be highly effective in the detection of antigens of this virus. Not differing in specificity from the direct TR-FIA, the newly developed test system was 4 times as sensitive.

[The differentiation of viruses in the Venezuelan equine encephalomyelitis complex by using monoclonal antibodies and lanthanide immunofluorescence analysis]. / Differentsiatsiia virusov kompleksa venesuél'skogo éntsefalomielita loshadei s primeneniem monoklonal'nykh antitel i lantanidnogo immunofliuorestsentnogo analiza.

Potentialities of differentiation between Venezuelan equine encephalomyelitis (VEE) complex viruses by time-resolved fluoroimmunoassay and enzyme immunoassay were studied. For this, 4 test systems were used based on different combinations of native and labeled polyclonal antibodies to VEE virus, strain Trinidad, and monoclonal (MCA) antibody MAK 14-7 to protein EL of this virus. The maximal sensitivity and specificity was achieved in the test system formed from native MCA MAK 14-7 for sensitization of the solid phase and labeled polyclonal immunoglobulins for demonstration of the test results. This combination of antibodies allowed to differentiate the epidemic variant of VEF/Trinidad (IA) from epizootic variants of Mucambo (III), Pixuna (IV) and attenuated strain No. 230.

[Monoclonal antibodies to the Karelian fever virus. The characteristics of their biological properties]. / Monoklonal'nye antitela k virusu karel'skoi likhoradki. Kharakteristika biologicheskikh svoistv.

Biological properties of 6 variants of monoclonal antibodies (MAb) to Karelian fever virus, a member of the alpha-virus serocomplex Sindbis-WEE, produced by the available hybridomas. The productivity of hybridomas of the "Karel" series in tissue culture and in cultivation as ascitic fluid was evaluated. Among the antibodies analysed, all were specific to envelope proteins, of them 2 were against protein E2 and four against protein E1. Comparison of MCA biologic activity (neutralizing, antihemagglutinating activities, participation in immunofluorescence, EIA, and immune blotting) allows one to distinguish four different hybridomas among them producing specific antibodies differing in their properties.

[Monoclonal antibodies cross-reacting with the tick-borne encephalitis virus and with the Venezuelan equine encephalomyelitis virus]. / Monoklonal'nye antitela, perekrestno reagiruiushchie s virusom kleshchevogo éntsefalita i virusom venesuél'skogo éntsfalomielita loshadei.

Employment of radioimmunoassay led to the demonstration of serological crossing between tick-borne encephalitis (TBE) virus and Venezuelan equine encephalomyelitis (VEE) virus. Using hybridoma technology, three hybridomas were produced secreting monoclonal antibodies (MAb) cross-reacting with these two viruses. With MAb, the epitope of binding of these antibodies was shown to be located on protein E of TBE virus and protein E1 of VEE virus. Despite the low percentage (14%) of homology of amino acid sequences of these proteins, 12 areas with homology from 24% to 63% were demonstrated. Considering conservative replacements, homology of these areas was 53%-75%. The assumed existence of some of these areas in alpha-helical conformation may explain the observed immunological crossing.

[Virological study of cases of sandfly fever in Afghanistan]. / Virusologicheskoe izuchenie sluchaev moskitnykh likhoradok v Afganistane.

Three strains (Af-1008, Af-1038, and Af-130) of Neapolitan sandfly fever virus (NSFF) and 2 strains (Af-1028 and Af-83) of Sicilian sandfly fever virus (SSFF) were isolated from febrile patients among soldiers of the limited military contingent of Soviet troops in Afghanistan in May-August, 1986-1987. The viruses were isolated in neonate mice and identified by indirect immunofluorescence, complement-fixation tests and plaque reduction neutralization test in Vero E6 and SPEV cells. The new NSFF strains in serological tests differed slightly from the original Sabin strain and completely differed from Toskana virus. In the examinations of 104 paired sera from convalescents, 87 subjects showed a rise in antibody titres either to NSFF or to SSFF virus, and 16 patients had antibodies to both viruses but without any rises in titre. Among 60 subjects with a presumable diagnosis of sandfly fever 12 persons had an antibody No information on isolation of sandfly fever viruses in Afghanistan has been available hitherto.

[Antigenic analysis of tick-borne encephalitis virus group using monoclonal antibodies and immunofluorescence]. / Antigennyi analiz virusov gruppy kleshchevogo éntsefalita s ispol'zovaniem monoklonal'nykh antitel i immunofliuorestsentsii.

The study included 18 monoclonal antibodies (MAb) to E- or NS1-antigens tested by immunofluorescence with tick-borne encephalitis (TBE) complex viruses. MAb were induced to 3 strains of TBE virus: the pathogenic 4072 strain isolated from a patient; the Skalica strain of low pathogenicity; and the Neidorf strain isolated from ticks. According to their reactivity to complex viruses, MAb comprised 3 groups: monospecific for TBE virus (T6, T15) which detected tick-borne encephalitis virus alone; widely cross-reactive with 4-6 viruses of the complex (NEK, KEN, T7, T9); and partially complex-reactive (T11, T12, T13, T33/3) and bound to 2-3 viruses of the complex. T13 and T33/3 MAb reacted with the Omsk hemorrhagic fever virus to the same degree or stronger than with TBE virus. The cross-reactivity was more marked in anti-E-than in anti-NS1 MAb. The similarity of the Langat viruses and the Skalica strain was confirmed. Using anti-NS1 MAb in tests with non-fixed cells, the release of NS1-antigen was found to begin at hour 18 (time of observation). The results of the study may be useful for improvement of laboratory diagnosis of TBE and evaluation of the capacity of a vaccine to induce cross immunity to viruses of the TBE complex.

[Hybridomas producing monoclonal antibodies to Crimean hemorrhagic fever virus]. / Gibridomy, produtsiruiushchie monoklonal'nye antitela k virusu Krymskoi gemorrhagicheskoi likhoradki.

A series of hybridomas producing monoclonal antibodies to Crimean hemorrhagic fever virus was obtained and their cultural properties were characterized. HEMA-12 and HEMA-24 secreted monoclonal IgG2b antibodies, HEMA-101 secreted monoclonal IgG1 antibodies, HEMA-31, HEMA-9 and HEMA-11 secreted monoclonal IgG2 antibodies. According to the results of the indirect immunofluorescence test, the titer of specific immunoglobulins in the culture fluid was 1:16-1:32, but sometimes reached 1:64-1:128. The titer of antibodies in ascitic fluid amounted to dilutions of 10(4)-10(5). Hybridomas were cloned by the method of ultimate dilutions. The injection of 5-15 million HEMA cells into the abdominal cavity of BALB/c mice induced the formation of ascitic tumors in the animals within 7-11 days and the accumulation of ascitic fluid in a volume of 1-4 ml. Hybridomas, found to be capable of passage from mouse to mouse, underwent 5-16 passages during the term of observation.

[Characteristics of the monoclonal antibodies induced by the Crimean hemorrhagic fever virus]. / Kharakteristika monoklonal'nykh antitel, indutsirovannykh virusom krymskoi gemorragicheskoi likhoradki.

Twelve lines of hybridomas secreting monoclonal antibodies (MCA) have been generated by fusion of spleen lymphocytes of BALB/c mice immunized with crude Crimean hemorrhagic fever virus (CHF). The hybridomas multiply actively in vitro (over 50 passages) and as ascitic tumors in the abdominal cavity of BALB/c mice. MCA were characterized by indirect immunofluorescence (IF), complement fixation (CF), biological neutralization test (NT), agar gel diffuse precipitation tests, and by the type of immunoglobulins. In the indirect IF, all kinds of MCA reacted with CHF virus, in CF only GEMA 12 and GEMA 31 which also precipitated the virus antigen in AGDPT. The CF-positive MCA belonged to the IgG2a and IgG2b subclasses, but 2 more MCA species of the same immunoglobulin type did not react in CF. No neutralizing properties have been found with MCA. All MCA reacted similarly in indirect IF and CF both with CHF and Congo viruses. Among other members of the CHF-Congo antigenic group, only GEMA 4 MCA interacted with Hazara virus. MCA generated in this study are intended for use in identification of CHF and Congo viruses in diagnostic tests.

[Monoclonal antibodies to flavivirus antigens induced by a vaccinal strain of yellow fever]. / Monoklonal'nye antitela k flavivirusnym antigenam, indutsirovannye vaktsinnym shtammom zheltoi likhoradki.

Monoclonal antibodies (MABs) YEL-2 induced by the vaccine FNS Dakar yellow fever (YF) virus were characterized for their capacity to enter into serological reactions and to react with heterologous flaviviruses. YEL-2 MABs belong to the IgG2a class of immunoglobulins, possess the antihemagglutination properties, are active in indirect IF test but do not activate complement and have no neutralizing properties. The inability to enter into CFT in the presence of antihemagglutinating properties suggests that YEL-2 MABs are directed for the structural E glycoprotein. YEL-2 MABs reacted similarly with the vaccine 17D strain and the FNS Dakar strain by which they had been induced. In addition to YF virus, YEL-2 MABs reacted with Tyuleniy, Rosio, Ilhéus, Uganda S, Karshi, and Sokuluk viruses, the reaction with Tyuleniy virus reaching the same titer as with the homologous virus but was of one-way nature. No reaction of YEL-2 MABs was observed with the viruses of the tick-borne encephalitis complex, Japanese encephalitis, West Nile, Dengue 2 and 4 viruses. These results specify the antigenic classification of flaviviruses.

[Monoclonal antibodies to the yellow fever virus]. / Monoklonal'nye antitela k virusu zheltoi likhoradki.

A highly technological Yel-2 hybrid cell clone was isolated. The clone produces monoclonal antibodies to protein E of the yellow fever virus (Dakar strain). The Yel-2 hybridoma was cloned by the method of limiting dilutions. By the data of indirect immunofluorescence the titre of the specific antibodies reached 1:128 in the culture fluid and 1:10240 in the ascitic fluid. The Yel-2 antibodies had no cross reactions with forest-spring encephalitis, Dengue 2 and Dengue 4 viruses. In the hemagglutination inhibition test the activity of the Yel-2 ascitic preparations was observed at a dilution of 1:50000 while the complement fixation test did not reveal it. Administration of the Yel-2 cells into the abdominal cavity of mice BALB/c induced development of ascitic tumors in 97 per cent of pristane-sensitized mice and in 90 per cent of nonsensitized mice. The hybridoma was successfully transplanted from mouse to mouse (more than 20 passages during the observation period) and maintained high constant levels of specific antibody secretion. The high ascite forming capacity of the Yel-2 hybridoma provided decreasing of the cell dose administered to the mice from 10(7) to 10(4) cells per mouse.

[Conditions for forming ascitic tumors in hybridoma cultivation in vivo]. / Izuchenie uslovii obrazovaniia astsitnykh opukholei pri kul'tivirovanii gibridom in vivo.

The conditions of the formation of ascitic cells in BALB/c mice injected with hybridoma cells were studied. All the hybridomas under study, producing monoclonal antibodies to viral antigens, induced the formation of ascitic tumors when introduced into the abdominal cavity of BALB/c mice pretreated with sensitizing agents. In the mice pretreated with pristane hybridoma cells took at a rate of 43-80% and in the mice pretreated with Freund's complete adjuvant, 31-70%. Angara oil and perfume oil, as well as Bayol F, were less effective. The time of the formation of ascites was inversely proportional to the dose of the injected cells, while the volume of ascitic fluid depended rather on the type of hybridoma and not on the dose of the injected cells. The study showed that the use of physiological saline or culture medium without serum for washing the abdominal cavity of mice after withdrawing ascites permitted the additional collection of 2.6-13.7 million hybrid cells, as well as a considerable amount of immunoglobulins.

[Antiviral activity of proteinase inhibitors in cultured cells infected with alpha-viruses]. / Antivirusnoe deistvie ingibitorov proteinaz v kul'ture kletok, zarazhennykh al'favirusami.

The ability of synthetic inhibitors of trypsin-like (TLCK) and chymotrypsin-like (TPCK) proteinases and natural antiproteinase oligopeptides of animal (aprotinin) and microbial (enzistatin) origin to suppress multicycle replication of different alpha viruses (Semliki, Sindbis, Venezuelan equine encephalomyelitis viruses) in cultured cells was studied. Antiviral activity was found to be induced by TPCK and aprotinin (Gordox). These compounds were shown to reduce virus yield 100-fold and to prevent the involvement of cells into infection process. The mechanisms of antiviral activity and chemotherapeutic possibilities of antiproteinase compounds are discussed.

[Serological characteristics of KAMA-51 monoclonal antibodies to the tick-borne encephalitis virus]. / Serologicheskaia kharakteristika monoklonal'nykh antitel KAMA-51 k virusu kleshchevogo éntsefalita.

KAMA-51 monoclonal antibodies to tick-borne encephalitis virus are produced by hybridoma obtained by fusion of splenocytes of mice immunized with this virus with myeloma X-653 cells. The antibody belongs to the IgG class, is active in the immunofluorescence test (IFT), does not react in CFT, and has no antihemagglutinating or neutralizing properties. In the IFT, it reacts with all viruses of the tick-borne encephalitis complex indicating their directivity to the groupspecific determinant of E protein. In the indirect IFT, antibody titres in the culture fluid are within the range of 1: 16-1: 62, in the ascitic fluids 1: 320-1: 640. Because of the wide range of interspecies reactions, KAMA-51 monoclonal antibody may be used for group detection of the tick-borne encephalitis complex viruses.