[Development of a rapid method, based on the latex agglutination reaction, for diagnosing carnivore plaque]. / Razrabotka methoda ékspress-diagnositki chumy plotoiadnykh na osnove reaktsii lateksnoi aggliutinatsii.

A highly effective rapid method for detecting carnivore plague virus antigen in dogs, based on the latex agglutination test, is developed. Latex conjugate is synthesized on the base of polystyrene suspension, representing a copolymer of styrene and methacrylic acid. The immunospecific reagent is gamma-globulin fraction of rabbit serum containing antibodies specific to carnivore plague virus. The resultant latex conjugate can be used in practical veterinary as a diagnostic agent for screening dogs for carnivore plague virus.

[Antigenic activity of measles immunostimulating complex]. / Antigennaia aktivnost' korevogo immunostimuliruiushchego kompleksa.

The liposomal technology for preparing the immunostimulating complex (ISCOM) helped obtain a complex measles preparation whose antigens are represented by the structural proteins of measles virus in the bilayer phosphate-lipid membrane. Immunization of mice with the resultant preparation induced antimeasles antibodies with the maximum titer of antihemagglutinins 1:640 and of antihemolysins 1:1280. The biological activity of antibodies was confirmed in the neutralization test.

[Immunoenzyme test systems for detecting the HIV antigen and a trial of their use in examining HIV-infected subjects]. / Immunofermentnye test-sistemy dlia vyiavleniia antigena VICh i popytka ikh ispol'zovaniia pri obsledovanii VICh-infitsirovannykh lits.

The results of the development and trials of two variants of EIA test system for the detection of HIV-1 antigen using biotinated HIV-antibody and streptavidin-peroxidase or biotin-beta-lactamase enzyme complexes are presented. These diagnostic preparations proved to be highly sensitive and specific allowing the detection of the antigen in the blood of HIV-infected subjects.

[Preparation of immunoenzyme conjugates of beta-lactamase from Bacillus licheniformis 749/c and horseradish peroxidase with human antibodies to HIV-1]. / Poluchenie immunofermentnykh kon"iugatov beta-laktamazy iz Bacillus licheniformis 749/c i peroksidazy khrena s chelovecheskimi antitelami k VICh-1.

By using three different linkage methods with carbodiimide, glutaraldehyde and periodite, immunoenzyme conjugates of beta-lactamase from Bacillus licheniformis 749/c and horse radish peroxidase with human antibodies to HIV-1 were prepared. The human antibodies were purified by the affinity procedure on Protein-A-Sepharose 6B. The conjugates were tested in a solid phase immunoenzymatic system for the HIV-1 antigen. It was shown that the conjugates prepared by the carbodiimide linkage method had the highest titer, the beta-lactamase conjugate being superior by its titer to the respective peroxidase conjugate. In the lyophilized state the conjugates prepared with the carbodiimide linkage method were stable.

[Reverse transcriptase of the human immunodeficiency virus: cloning, expression in Escherichia coli, purification of the enzyme, and production of monoclonal antibodies]. / Obratnaia transkriptaza virusa immunodefitsita cheloveka: klonirovanie, ékspressiia v Escherichia coli, ochistka fermenta i poluchenie monoklonal'nykh antitel.

To express HIV-1 reverse transcriptase in E. coli a number of genetic constructions containing reverse transcriptase and virus protease nucleotide sequences was obtained. The products of expression were characterized; monoclonal antibodies to reverse transcriptase were produced. The purification of reverse transcriptase was carried out. The substantial proteolysis of reverse transcriptase during purification was shown. The purified preparation is predominantly, an active protein with Mr 57 kDa. Some properties of this protein differed from the reverse transcriptase isolated from HIV.

[Biosynthesis of the membrane protein of the acquired immunodeficiency syndrome virus (HIV) with a deleted hydrophobic region in E. coli cells]. / Biosintez obolochechnogo elka virusa sindroma priobretennogo immunodefitsita (HIV) s udalennoi gidrofobnoi oblast'iu v kletkakh E. coli.

Effect of the structure of cloned env gene of HIV virus on the level of its expression has been studied. The deletion of the hydrophobic region from the env gene has been shown to increase considerably the biosynthesis of antigen-specific proteins in Escherichia coli cells. At the same time, the low level of expression of the gene fragment the transcription of which is controlled by a powerful lac-promoter suggests the presence of toxic regions of nonhydrophobic nature in the synthesized antigen.

Interaction of influenza virus with gangliosides and liposomes containing gangliosides.

It has already been shown that influenza virus binds unspecifically to liposomes containing ganglioside GM1 wheras with gangliosides GD1b and GT1b binding occurs in a specific and saturable manner [Slepushkin et al. (1986) Biol. Membr. 3, 229-235]. In the present study the mode of interaction between influenza virus and various gangliosides or phospholipid liposomes containing cholesterol and gangliosides has been investigated. The influence of exogenous gangliosides on the structure of the viral envelope was studied using fluorescent and photoactivatable phospholipids incorporated into the viral membrane. With both types of probes maximal effects of gangliosides were caused by GT1b. Addition of that ganglioside resulted in a marked decrease in the fluorescence polarization (P) of fluorescent labeled virus as well as in substantial changes in the binding of photoactivatable analogues of sphingomyelin and phosphatidylcholine to virus proteins, mainly hemagglutinin. The effects of GT1b and GD1b on P value were comparable, whereas gangliosides with other oligosaccharide chains caused much smaller changes in P. Furthermore GT1b but not GM1 influenced phospholipid-hemagglutinin cross-linking. Interaction of the virus with large unilamellar liposomes was monitored by two fluorescence assays based on resonance-energy transfer from the tryptophans and tyrosines of viral proteins to vesicles labeled with a triacylglycerol (anthrylvinyldioleoylglycerol) or from these labeled vesicles to virions labeled with a perylenoyl derivative of galactosylcerebroside (PGalSph). A third fluorescence assay was based on relief of self-quenching in PGalSph-labeled virions, upon low-pH-induced virus-liposome fusion. With all three fusion assays the changes of fluorescence caused by GT1b were more pronounced than those induced by GM1. On the other hand, virus-induced release of [14C]glucose from multilamellar liposomes was enhanced by GM1 but not by GT1b or GD1b. It is concluded that the interaction of GT1b or GD1b with virus hemagglutinin induces a rearrangement of the viral lipids rendering lipid bilayer areas of the viral envelope significantly fluid, which in turn promotes fusion of the virus with target membranes. Probably virus-liposome fusion and virus-induced liposome leakage are brought about by different mechanisms depending on specific or unspecific binding of the virions to the target.

[Differences in the gene structure of the matrix protein and hemagglutinin in remantadine-resistant and -sensitive variants of the influenza virus type A FPV]. / Razlichiia v strukture genov matrikskogo belka i gemaggliutinina ustoichivogo i chuvstvitel'nogo k remantadinu variantov virusa grippa tipa FPV.

The oligonucleotide mapping technique and RNA-RNA hybridization revealed structural differences between the HA- and M-protein genes of two variants of influenza A FPV, remantadine-sensitive and remantadine-resistant ones. A quantitative estimation of the structural divergence of the two influenza FPV pairs of genes was carried out. The possible functional role of the differences in the structure of the HA- and M-protein genes of the two FPV variants is discussed.

[Rapid diagnosis of rotavirus infections by RNA electrophoresis on polyacrylamide gel]. / Bystraia diagnostika rotavirusnykh infektsii putem élektroforeza RNK v poliakrilamidnom gele.

A method of rapid diagnosis of rotavirus infections is described. It is based on the isolation of viral RNA from the feces of patients with viral gastroenteritis and determination of RNA mobility by 6% polyacrylamide gel electrophoresis. It was demonstrated that rotavirus infection could be diagnosed after long-term storage of fecal specimens as well. The method is of interest for epidemiological and genetic studies of human rotavirus diseases.

[Morphological characteristics of the rotaviruses and the structure of their genome]. / Morfologicheskaia kharakteristika rotavirusov i struktura ikh genoma.

The data on human and animal rotavirus reproduction in monkey kidney cell culture, their morphology, and genome structure are presented. Both viruses, simian rotavirus SA 11 and human rotavirus K1, did not differ morphologically, in both cases two-capsid and one-capsid particles were found. The genome structure was determined by electrophoresis in 6% PAG. The electrophoretic mobility of genome fragments of both viruses was similar.

[Isolation and characteristics of endocytic vacuoles containing the influenza virus]. / Vydelenie i kharakteristika éndotsitarnykh vakuolei, soderzhashchikh virus grippa.

Endocytic vacuoles (receptosomes) containing influenza virus were isolated from the cytoplasm of Ehrlich ascitic carcinoma cells and characterized. In the sucrose density gradient, the virus-containing material was detected in two peaks with a buoyant density of 1.175-1.16 and 1.155-1.135 g/cm3 with which the activity of marker enzymes of cell plasma membranes was associated. The virus was present in receptosomes in morphologically and electrophoretically intact condition. Examinations for the lipid composition of endocytic vacuoles showed the presence in their membranes of large amounts of cholesterol and glycolipids, particularly asialo-GM1 which, according to some authors may enhance the fusion of viral and cell membranes.

[Transcription of influenza virus genome. Location and dependence on the nuclear primer function]. / Transkriptsiia genoma virusa grippa. Lokalizatsiia i zavisimost' ot praimernoi funktsii iadra.

The synthesis of the complete and incomplete transcripts (the templates in genome replication and mRNA, respectively) of influenza A WSN virus in chicken fibroblasts was analyzed by gel electrophoresis analysis of the duplexes formed between virion RNA and complementary RNA. Three steps in the transcription could be defined: 1) primary transcription when similar amounts of mRNA of all the genes are accumulated; 2) early secondary transcription when mRNA of NS gene is synthesized in larger amounts than that of other genes and 3) late secondary transcription when the amplification of transcription from all the genes is performed. The synthesis of complete transcripts starts during or after primary transcription. When actinomycin D was added to infected cells, the synthesis of incomplete transcripts was inhibited to a larger degree then that of complete transcripts. Most of incomplete transcripts was observed within the cell nucleic while the complete transcripts were found in the nuclei and cytoplasm, suggesting that the synthesis of incomplete transcripts is located in the nuclei. alpha-Amanitin blocked the synthesis of incomplete transcripts without interfering with that of complete transcripts. These data suggest that the synthesis of complete transcripts does not require the synthesis of cell mRNA as primers in transcription.

[Differences in the biosynthesis of 2 types of viral transcript in cells infected with influenza virus]. / Razlichie biosinteza dvukh tipov virusnykh transkriptov v zarashennykh virusom grippa kletkakh.

An analysis of poly A(+)-and poly A(--)-transcripts in influenza A/WSN (H0N1) virus-infected cells done by the method of Hay and coworkers demonstrated three stages of transcription: (1) primary transcription during which mRNA of all the genes is synthesized, (2) early secondary transcription during which mRNA of NS gene is amplified, and (3) late secondary transcription in which mRNA of all the genes is amplified. Poly A(--) transcripts are synthesized at the stage of secondary transcription. The effect of three concentrations of actinomycin D at various stages of transcription was studied. The experimental results suggest differences in inhibition by actinomycin D of synthesis of transcripts of both types at the early and late stages of secondary transcription. The effect on the infected cells of alpha-amanitine specifically inhibiting synthesis of primary mRNA showed the synthesis of cellular RNAs to be much more necessary for the synthesis of polyA(+)-transcripts than for that of polyA(--)-transcripts.