[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.

[Identification of monoclonal antibodies against protein E1 of venezuelan equine encephalomyelitis virus blocking the hemagglutinating but not the infective activity of virions]. / Identifikatsiia klona antitel k belku E1 virusa venesuél'skogo éntsefalomielita loshadei, blokiruiushchego gemaggliutiniruiushcuiu, no ne infektsionnuiu aktivnost' virionov.

Cellular clone (MAK 14-7), producing antibodies against the virus of Venezuela equine encephalomyelitis (VEE), strain 230, was isolated using the standard hybridomata technology. Monoclonal antibodies neutralized the viral hemagglutinating activity leaving the infectious one intact. Monoclonal antibodies from MAD 14-7 reacted specifically with viral glycoprotein E1 as registered by the immunoprecipitation technique. The topography of antigenic determinants of viral E1/E2 glycoprotein dimer forming the virions outer spikes is discussed in connection with the results obtained.

[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.

[Radial hemolysis reaction with arboviruses]. / Reaktsiia radial'nogo gemoliza s arbovirusami.

A modification of the radial hemolysis test with arboviruses of the antigenic groups A and B was developed with Sindbis, Chikungunya, Venezuelan equine encephalomyelitis, West Nile, dengue 1 and 2 viruses. Goose erythrocytes were sensitized with hemagglutinating sucrose-acetone antigens of these viruses from mouse brains at pH optimal for each virus.

[Preparation of ribonucleoproteins and virion envelopes of the Zaisan-260 and R-16225 strains of Semliki Forest virus antibody production]. / Poluchenie ribonukleoproteidov i obolochek virionov shtammov Zaisan-260 i R-16225 virusa lesa Semliki dlia polucheniia antitel.

Conditions for the preparation of subviral structures, envelope and ribonucleoprotein, of the Zaisan-260 and R-16225 strains of Semliki Forest virus inducing antibody response in rabbits inoculated into lymph nodes were developed. The virus was cultivated in chick embryo fibroblasts in a medium containing rabbit serum. Ribonucleoproteins and envelopes were isolated by virus disruption with Nonidet P-40 at 25 degrees C for 20 min followed by fractionation in a linear sucrose density gradient 15-30% at 150,000 X g for 50 min, and identification of envelopes and ribonculeoprotein by 14C and 3H labels. Rabbit immune sera to virions, envelope and nucleopsid fragments of the both Semliki Forest strains wer prepared. The immune sera to the intact virion had slightly higher antibody titres than sera to subunits.

[Alphavirus reproduction in cultures of transplantable human lymphoblastic cells in acute infection]. / Reproduktsiia al'favirusov v kul'turakh perevivaemykh limfoblastoidnykh kletok cheloveka pri ostroi infektsii.

The results of investigations of acute infection of continuous human B- and T-cells with typical members of the alphavirus group--Semliki Forest, Sindbis, and Venezuelan equine encephalomyelitis viruses, are presented. Virus amplification was shown to pass through the typical phases: eclipse, logarithmic growth, plateau. Infectious virus production per one cell was from 10 to 10,000 PFU in various cultures. Cell infection results in interferon production. Replication of the viruses under study in lymphoblastoid cell cultures is not accompanied by the active cytocidal effect. The regularities determining the sensitivity of lymphoblastoid cells to viruses in general and alphaviruses in particular are discussed. Proceeding from the results of the study of alphavirus replication in human continuous B- and T-cells it is suggested that this system be used as a model for the analysis of antiviral activity of interferon, its inducers, and chemopreparations in special cells. Lymphoblastoid and fibroblast interferon are as active in B-cells and show no antiviral activity in continuous T-cells, as interferon inducer.

[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.

[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.

[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.

[Isolation of glycoproteins of the Venezuelan equine encephalomyelitis virus and an evaluation of their immunogenic activity]. / Vydelenie glikoproteidov virusa venesuél'skogo éntsefalomielita loshadei i otsenka ikh immunogennoi aktivnosti.

Isolation of purified Venezuelan equine encephalomyelitis virus glycoproteins by means of a new Soviet nonionic detergent, MESK, is described. The MESK detergent was shown to permit isolation of approximately 70% of virus particle glycoproteins. The resulting preparation had a high hemagglutinating activity, contained no admixtures of foreign proteins and was not infectious. The study of the immunogenicity of purified glycoproteins in experimental mice and rabbits showed them to be capable of inducing high levels of serum antibodies. The immunogenicity of the isolated glycoproteins was comparable to their immunogenicity as components of virus particles. Treatment of the virus with MESK detergent also yielded preparations with predominant content of capsid protein. The described procedure for disintegration and purification of viral proteins is technologically simple and may be the basis for manufacture of a subunit vaccine. The resulting material may also be used for preparation of diagnosticums and in laboratory studies.

[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.

[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.

[Identification of a group A arbovirus isolated in the Far East]. / Identifikatsiia arbovirusa iz gruppy A, vydelennogo na Dal'nem Vostoke

Strains Nos. 296 and 260 isolated from Aedes vexans and Culex pipiens mosquitoes in the USSR Far East in 1971 and 1973, respectively, were identified. The size of the viruses was found to be about 50 nm. Both strains were pathogenic for 2--3-day-old and 3--4-week-old mice by the intracerebral, subcutaneous and intraperitoneal routes, agglutinated goose red blood cells at the optimal pH 6.0. The strains were identical to each other and by their antigenic structure were classified as Semliki Forest virus.

[Antiviral action of monoclonal antibodies]. / Protivovirusnoe deistvie monoklonal'nykh antitel.

The antiviral effect of monoclonal antibodies MAK-14-7 possessing the neutralizing activity was studied on the model of Venezuelan equine encephalomyelitis virus in cell cultures and in experimentally infected laboratory animals. Mouse monoclonal antibodies exerted a marked specific antiviral effect in tissue culture and in white mice inhibiting virus reproduction by 1-3-5 log PFU/ml. The degree of inhibition of virus reproduction is inversely proportional to the multiplicity of infection. When used in combination with remantadine, ribavirin, interferon, and its inducer poly (I) X poly (C), the monoclonal antibodies MAK-14-7 exerted additive antiviral effect.

[Radial hemolysis reaction in the diagnosis of tick-borne encephalitis]. / Reaktsiia radial'nogo gemoliza v diagnostike kleshchevogo éntsefalita.

A test of radial hemolysis in gel (RHG) has been developed and first used for serodiagnosis of tick-borne encephalitis (TBE). In examinations of 464 blood serum specimens from 258 patients with TBE and subjects suspected of this disease in RHG and HI tests the diagnosis of tick-borne encephalitis was confirmed in 77 subjects, the results of both tests being in complete agreement. A comparative analysis of antibody levels by RHG and HI tests revealed a complete correlation. An advantage of RHG over HI test is its insensitivity to serum inhibitors and the possibility of quantitation of antibodies in whole sera which omits a complicated procedure of serum treatment to remove inhibitors and serum titrations in serial dilutions mandatory for HI tests. The specificity and sensitivity of RHG, simplicity in running and the possibility of analysing many sera within a short time recommend the RHG test for public health practice for TBE serodiagnosis.

[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.

[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.

[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.

[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.