Vol 62, No 3 (2017)

ORIGINAL RESEARCH

MOLECULAR-GENETIC CHARACTERIZATION OF FIELD ISOLATES OF RABIES VIRUS IDENTIFIED IN THE TERRITORY OF VLADIMIR, MOSCOW, TVER, NIZHNY NOVGOROD AND RYAZAN REGIONS

Zaykova O.N., Grebennikova T.V., Gulyukin A.M., Shabeykin A.A., Polyakova I.V., Metlin A.E.

Abstract

The article presents a molecular genetic study of genomes of field isolates of rabies virus isolated in the Vladimir, Moscow, Tver, Nizhny Novgorod and Ryazan regions, with the aim of carrying out phylogenetic analysis. We studied 20 samples of purified PCR products containing the rabies virus nucleoprotein. The samples were provided by the Vladimir veterinary service. Sequencing and phylogenetic analysis of the gene showed that 12 fragments of isolates under study were close to the Central phylogenetic group of the rabies virus; namely - 5 isolates from the Vladimir region, 2 from the Nizhny Novgorod region, 2 from the Moscow region, and 3 from the Tver region. Eight studied isolates from the Nizhny Novgorod and Ryazan regions were attributed to the Eurasian phylogenetic group.
Problems of Virology. 2017;62(3):101-108
pages 101-108 views

DIAGNOSTIC CAPACITY OF DETECTION OF SPECIFIC ANTIBODIES TO PANDEMIC INFLUENZA A(H1N1)PDM09 VIRUS

Mukasheva E.A., Nikolaeva L.I., Makhnovsky P.I., Kirillova E.S., Kolobukhina L.V., Merkulova L.N., Kruzhkova I.S., Malyshev N.A., Burtseva E.I.

Abstract

Serologic studies occupy a significant place in influenza diagnosis. The article presents an analysis of the developed experimental version of ELISA test-systems for the detection of specific antibodies to the virus influenza A(H1N1)pdm09, and their dynamics at different stages of infection as compared with those of the traditional HAI method. The study included 20 paired samples of serum from patients hospitalized at different stages of the disease with etiology associated with the influenza virus A(H1N1)pdm09. Two groups were formed on the basis of HAI data, which showed the presence or absence of significant growth of specific antibodies to the influenza virus A(H1N1)pdm09. The control group consisted of 20 serum samples from individuals without influenza but with chronic hepatitis C. To examine the virus specific antibody two types of ELISA test systems were used. The first system was intended for the detection of IgM to the influenza virus A(H1N1)pdm09; the second was used for revealing specific IgG. The study showed the accuracy and specificity of detectable IgM and IgG to the virus influenza A(H1N1)pdm09. The dynamics of specific IgG titers in 15 of the 20 pairs of sera was reliable. The increase in titers was more pronounced than in the HAI. IgM against influenza virus could be detected up to 10 days, although reliable dynamics of these antibodies was not detected in paired samples. The test system was specific for the determination of both IgG and IgM antibodies to the influenza virus A(H1N1)pdm09 and significantly more sensitive than HAI. Using this ELISA test system, it is possible to monitor the dynamics of IgG to this virus even in the absence of diagnostic increases in antibody titers in HAI.
Problems of Virology. 2017;62(3):109-114
pages 109-114 views

THE INFLUENCE OF HIV INFECTION DURATION BEFORE ANTIRETROVIRAL THERAPY ON IMMUNOLOGICAL TREATMENT FAILURE

Oleynik A.F., Fazylov V.K.

Abstract

Introduction. Some patients with HIV infection receiving virologically effective antiretroviral therapy (ART) did not show any growth in CD4 cell count during treatment. Despite the long-term treatment of patients, immunodeficiency persisted. Methods. In the observational cohort retro/prospective study we investigated the effect of the duration of HIV infection before starting the antiretroviral therapy on the development of immunological treatment failure. Results. In a group of 140 HIV-infected patients a moderate inverse correlation was found between the duration of HIV infection and CD4 cell gain after 6 months, 1, 2 and 3 years of ART (r = -0.33, p < 0.01; r = -0.3, p < 0.01; r = -0.3, p < 0.01; r = -0.29, p < 0.01, respectively). In the case of ART starting at CD4 count of 200-350 cells/mcl statistically significant differences were revealed in the levels of relative CD4 count at 6 months, 1, 2 and 3 years of ART in sub-groups with durations of HIV infection prior to initiating therapy of 1-8 years and > 8 years (p = 0.035; p = 0.015; p = 0.05; p = 0.05, respectively). In patients who started ART with CD4 levels > 200 cells/mcl after 8 years of HIV infection, the risk of immunological treatment failure is 4 times higher as compared to patients with the same CD4 level, but lesser duration of HIV infection. Conclusions. The shorter the duration of HIV infection, the greater the increase in CD4 cell count. When starting ART at CD4 level of 200-350 cells/mcl, restoration of CD4 count is more intense in the group with lesser period of HIV infection. A negative effect of the >8-year infection duration before the start of ART on the development of the immunological treatment failure was observed in a subgroup with a starting level of CD4 > 200 cells /mcl.
Problems of Virology. 2017;62(3):114-119
pages 114-119 views

MORPHOLOGICAL ANALYSIS OF HEPATITIS B VIRUS WITH ESCAPE MUTATIONS IN S-gene G145R AND S143L

Konopleva M.V., Sokolova M.V., Shevlyagina N.V., Bazhenov A.I., Fel’Dsherova A.A., Krymskij M.A., Borisova V.N., Semenenko T.A., Nesterenko V.G., Suslov A.P.

Abstract

Background. In terms of serological properties and immunization, the wild type of HBsAg HBV and its G145R mutant behave as different antigens. This testifies to serious structural changes, which presumably could have a significant impact on the morphogenesis of virions and subviral particles. Nevertheless, morphological and ultrastructural investigations of HBV with G145R mutation have not been carried yet. Objectives. Research of structural and morphological organization of HBV in the presence of the G145R escape mutation. Methods. Studies of sera, purified viruses and recombinant HBsAg were carried out by transmission electron microscopy by the method of negative staining and indirect reaction of immunelabeling using monoclonal antibodies of different specificity. Specimens of wild type HBV and HBV with S143L mutation obtained in an identical manner were used as the control. Results. The presence of typical virus particles of HBV was shown in the specimens of wild strain and HBV with S143L mutation. Specimens of HBV with G145R mutation were characterized by expressed morphological heterogeneity. In the initial serum and in the specimen of purified virus containing G145R mutant, large oval particles 60-70 nm and up to 200 nm in size, respectively, were found. The presence of antigen structures of HBV in all heterogeneous forms was confirmed. It was shown that forming of subviral particles in the process of expression of the recombinant HBsAg with G145R mutation depends on conditions of expression and purification of the protein. They can vary from well-formed circular and oval particles to practically unstructured fine-grained masses. Conclusion. Direct data on the impact of G145R escape-mutation in S-gene, in contrast to S143L mutation, on the morphogenesis of virions and subviral particles of HBV were obtained.
Problems of Virology. 2017;62(3):119-128
pages 119-128 views

INFLUENCE OF THE IMIQUIMOD ON THE INTERFERON PRODUCTION AND TREATMENT OF THE EXPERIMENTAL HERPES SIMPLEX VIRUS INFECTION

Smirnov V.S., Petlenko S.V.

Abstract

Background. Imiquimod is an imidazole derivative acting as an immunomodulator on the level of innate and adaptive immune system. Our objective was to evaluate the antiviral activity of generically reproduced imiquimod administered subcutaneously in mice and intravaginally in guinea pigs against herpes simplex virus (HSV), as well as to study the dynamics of serum interferon (IFN) synthesis under different dosing regimens. Results. When administered subcutaneously at doses of 0.5, 1.0 and 10 mg/kg imiquimod increased IFN production in mice in a dose-dependent manner with maximum serum IFN concentrations occurring 4 hours after dosing. Imiquimod protected mice from intraperitoneal HSV infection at doses of 3.2 and 32 LD50.The utmost protection (100% survival) was observed when imiquimod was administered at a dose of 100 mg/kg daily for 5 days before infection. Topical application of imiquimod 5% cream exhibited significantly more rapid and complete virus elimination in guinea pigs intravaginally infected with HSV type 2 compared to control group. Conclusion. Imiquimod produced as a generic possesses the same immunomodulatory and antiviral properties as the originally synthesized substance.
Problems of Virology. 2017;62(3):128-134
pages 128-134 views

ADAPTATION OF THE CORN EARWORM SINGLE NUCLEOCAPCIDE NUCLEOPOLYHEDROVIRUS (HELICOVERPA ZEA SNPV) FOR THE CONTROL OF THE COTTON BOLLWORM (HELICOVERPA ARMIGERA) POPULATION

Kolosov A.V., Ternovoy V.A., Shvalov A.N., Moiseeva A.A., Safatov A.S., Mikheev V.N.

Abstract

Helicoverpa zea (Boddie, 1850) (Hz) single nucleocapcide nucleopolyhedrovirus (SNPV) was adapted to the cotton bollworm (Helicoverpa armigera, (Hübner, 1805) (Ha)) by five blind passages on larvae. The full genomic sequence of the resulting strain HS-18 has been determined (GenBank acc. №: KJ004000.1). Biological activity of the HS-18 strain is higher than the activity of all other Russian strains of NPV, as far as cotton bollworm strain HearSNPV-G4. HS-18-infected caterpillars at the 3-rd and 4-th ages died much faster than those infected with HearSNPV-G4 strain. A major difference of HS-18 genome is an 18 bp repeat in the RING-finger ORF that confirms high variability of this region. Three other insertions and seven base substitutions were observed earlier, while six base substitutions are new. Mutations are located at ORF42, lef-9, ORF58, VP39, PIF-4, P48, SOD, ORF111, ORF129 and ORF138 genes. Among all nucleotide mutation only one is synonymous. Thus we suppose the selective pressure to the virus. The resulting strain HS-18 is recommended as a biopesticide for controlling the number of cotton bollworm in cotton fields.
Problems of Virology. 2017;62(3):134-137
pages 134-137 views

TO VIROLOGIST’S AID

ADAPTATION OF HUMAN ROTAVIRUS STRAINS OF GROUP A TO THE REPRODUCTION IN PASSAGED CELL CULTURES

Kolpakov S.A., Kolpakova E.P.

Abstract

The incidence of rotavirus gastroenteritis in the world still has no tendency to reduction. The development of an effective vaccine would reduce or, in the future, even defeat this highly contagious dangerous disease. However, both in Russia and abroad there is still no developed technique for adapting and cultivating strains of the human rotavirus A that would stably produce a high "yield" of virus progeny in transplanted culture cells. The phenomenon of gene exchange for the segmented genome of rotavirus was used by foreign researchers to create the rotavirus vaccine using reassortant strains which are the result of joint cultivation of low-titer (1-2·106 virions per ml) human rotavirus strains and rotavirus strains of animals, such as monkey rotavirus SA-11 or Nebraska calf rotavirus diarrhea providing a relatively high "yield" of virus progeny (1·107-1·108). It is clear that such vaccine compositions will not be able to replace a full-fledged vaccine of human rotavirus strains of different serotypes, but they can be used for the time being as a solution to the problem. Ideally, a rotavirus vaccine is needed that includes the full set of G and P serotypes of rotaviruses circulating in the territory of their application. The paper describes an original technique for adaptation and cultivation of human rotaviruses of group A on the culture of transplantable cells developed by the authors. This technique allows 5·108 virions to be obtained per 1 ml of culture fluid. High-titer cultivated strains of human rotavirus that can be used as vaccine strains were obtained, as well as highly-active antigens for the construction of diagnostic test-systems.
Problems of Virology. 2017;62(3):138-143
pages 138-143 views

ANNIVERSARY DATES

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pages 144-144 views


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