Vol 64, No 3 (2019)
- Year: 2019
- Published: 20.06.2019
- Articles: 7
- URL: https://virusjour.crie.ru/jour/issue/view/30
- DOI: https://doi.org/10.36233/0507-4088-2018-64-3
Full Issue
REVIEWS
THE USE OF TRANSGENIC MOSQUITOES FOR PREVENTION OF SPREADOF ARBOVIRAL DISEASES
Abstract
The mosquitoes of Aedes genus are the most important vector such arboviral diseases as dengue, yellow, Chikungunya, West Nile and Zika fevers. Work is currently in progress to control the transmission of agents of these diseases by forming of transgenic mosquitoes in order to altering the capacity of wild mosquitoes to support of virus replication. There are two main strategies of genetic control of mosquitoes population. Sterile Insect Technique (SIT), that mainly uses population suppression methods for making self-sustaining genetic systems and Release of insects carrying of a Dominant Lethal (RIDL) that uses mainly gene transfer methods for making of self-limiting genetic systems. The RIDL is more expensive, but it has some significant preferences, according compares with SIT. The field trials of genetic control methods are conducted in several countries from 2009 to present time. Genetic control, transgenic technologies to induce sterility, genetic elimination and stable transformation of Aedes mosquitoes are viewed in this review.
Problems of Virology. 2019;64(3):101-104
101-104
ORIGINAL RESEARCH
IDENTIFICATION AND MOLECULAR-GENETIC CHARACTERISTICS OF THE HEPATITIS B VIRUS AMONG HIV-INFECTED PATIENTS IN ARKHANGELSK
Abstract
Aim. To estimate the prevalence and characterize the hepatitis B virus among HIV-infected patients with virological failure of antiretroviral therapy in Arkhangelsk. Material and methods. HBV markers determinations (HBsAg, anti-HBs IgG, anti-HBcor IgG, DNA HBV) were performed in isolates from blood plasma samples 64 HIV-infected patients with virological failure of antiretroviral therapy (viral load >50 IU / ml after 6 months of antiretroviral therapy or an increase in viral load after primary suppression of viral replication). For the detection of the hepatitis B virus, nucleic acids were isolated using the commercial kit «AmplePrime Ribo-prep». The virus presence analysis was performing by the polymerase chain reaction (PCR) method with hybridization-fluorescence detection in “real time” using the commercial set of «AmpliSens® HBV-FL». In the future, we used the method developed by the Saint-Petersburg Pasteur Institute, which allows detecting HBV in biological material with a low viral load. Results. HBsAg-negative (occult) HBV was detect in 28 (43.8%) HIV-infected patients. Only HBV genotype D was detected, and the HBV subgenotype D1 prevailed (39.3%) compared with the HBV subgenotype D2 (32.1%) and D3 (28.6%). Serological markers in 42.8% of patients with HBV DNA were founding. Two HBV isolates with drug resistance mutations in the polymerase gene, leaded to amino acid substitutions (L180M, M204V) associated with the resistance development to lamivudine, entecavir, telbivudine and tenofovir were identifying. Conclusion. The occult (HBsAg-negative) HBV high prevalence among HIV-infected patients suggests the need to use molecular-biological diagnostic methods to identify HBV, as well as to analyze the HBV drug resistance mutation before starting antiretroviral therapy for HIV.
Problems of Virology. 2019;64(3):105-111
105-111
DETECTION OF EPSTEIN-BARR VIRUS GENOME IN ORAL CAVITY SQUAMOUS CELL CARCINOMA SAMPLES OF RUSSIAN PATIENTS
Abstract
Introduction. Oral cavity squamous cell carcinoma (OC-SCC) is the most common and aggressive malignancy of the oral cavity. Recent studies have revealed infections with human papilloma virus (HPV) as an additional risk factor for oral squamous cell carcinoma development, while distinguished role of Epstein-Barr virus (EBV) remains still uncertain. However, the evidence for association between virus infection and risk of oral squamous cell carcinoma is controversially and varies significantly by geographic regions and race. Purpose. The aim of the present study was to elucidate the prevalence of HPV and EBV in OC-SCC samples of Russian patients from Moscow region. Material and methods. We investigated fresh-frozen tumor tissue fragments obtained from 11 patients with OC-SCC. DNA was extracted and the viral genome was examined by quantitative PCR assays with high-risk type-specific HPV and EBV specific markers followed by sequencing-based analysis. Results. No HPV infection in analyzed OC-SCC samples was observed, while EBV was identified in 70.0% (7/10) of patients. Further based on Q-PCR amplification of the EBV targets including BamHI-W, EBNA1 and C-terminal fragment of LMP1 gene, EBV infection and measurement of virus load in the tumor samples was assessed. Sequencing LMP1-positive products revealed that the most samples (5/6) contained variants LMP1 with Cao deletion characterized by an increased transforming potential. Conclusion. These data suggest that prevalence of EBV infections is common and may influence cancer development, although detected LMP1 variants of EBV are not necessarily associated with the pathogenesis of OC-SCC. Further studies are necessary to determine the potential role of EBV and its possible importance as an infection factor in OC-SCC.
Problems of Virology. 2019;64(3):112-117
112-117
ALLOKIN-ALPHA - NEW APPROACHES IN THE TREATMENT OF CHRONIC VIRUS EPSTEIN-BARR INFECTIONS
Abstract
Introduction. Epstein-Barr virus causes recurrent infectious mononucleosis-like symptoms. Today it is shown that the poisons of insects and animals are rich sources of antimicrobial substances (peptides) and contain a wide range of active biological compounds. Antimicrobial peptides play an important role in the immune response of the innate immunity of the host in the presence of pathogenic microorganisms. Russia has developed an antiviral drug Allokin-alpha on the basis of antimicrobial peptides. The active ingredient of this drug is cytokin-like peptide alloferon. The aim of the study is to evaluate the effect of allokin-alpha therapy on the amount of EBV DNA in saliva samples and clinical complaints in patients with chronic Epstein-Barr infection (ChEBVI). Material and methods. 59 patients with ChEBVI were examined (45 women and 14 men; mean age 32.52 ± 1.75 years). Patients were examined quantification of DNA Epstein-Barr virus in saliva samples by the method of polymerase chain reaction (PCR) with hybridization-fluorescence detection in “real time” mode. The analytical sensitivity of the test system is 400 copies / ml. Patients were randomized into two groups: group 1 (25 patients) received Allokin-alpha therapy (9 injections of s / c, 1.0 mg every other day); group 2 (33 patients) received Valtrex (500 mg x 2 times / day, by mouth) for two months. Results. 59.67% of patients had negative PCR results after treatment with Allocin-alpha. Only 27.27% of patients had negative PCR results after two months of treatment with Valtrex. In a correlation analysis, a significant effect of the initial number of copies of DNA EBV on the severity of clinical complaints in patients was revealed in the general group ChEBVI. Discussion. Allokin-alpha improves the recognition of virus-infected cells and helps suppress viral replication. Conclusions. Allocin-alpha therapy can be recommended for the treatment of chronic EBVI at a dose of 1 mg subcutaneously every other day with a course dose of at least 9 injections.
Problems of Virology. 2019;64(3):118-124
118-124
ANTIVIRAL EFFECT OF «KAGOCEL» SUBSTANCE IN VITRO ON INFLUENZA VIRUSES H1N1, H1N1PDM09 AND H3N2
Abstract
Introduction. Active circulation of pandemic influenza and new variants of influenza H3N2 strains requires monitoring of antiviral efficacy of drugs permitted for influenza therapy in the Russian Federation. Purpose. Assessment of antiviral efficacy of «Kagocel» substance against influenza viruses H1N1, H1N1pdm09 and H3N2 in vitro. Material and methods. Cytotoxic effect of «Kagocel» substance on MDCK cells had been determined by stained with MTS. Antiviral efficacy of «Kagocel» substance against influenza infection has been studied in vitro in the culture of MDCK cells infected with influenza virus strains: A/Puerto Rico/8/34 (H1N1), А/California/7/2009 (H1N1)pdm09, А/Hong Kong/1/68 (H3N2) and А/Hong Kong/4801/2014 (H3N2). The antiviral activity of «Kagocel» substance was tested by its effect on the infectious titer of the influenza viruses and on its impact on the expression level of viral antigens in the enzyme immunoassay test system. Results. «Kagocel» substance had low toxicity for MDCK cells. «Kagocel» inhibited the infection titer of influenza virus strains A/Puerto Rico/8/34 (H1N1), А/California/7/2009 (H1N1)pdm09, А/Hong Kong/1/68 (H3N2) and А/ Hong Kong /4801/2014 (H3N2) in the MDCK cell culture with equal efficacy. Study of the impact of «Kagocel» substance on the expression level of viral antigens by ELISA also revealed its antiviral efficacy for all tested strains. Dose dependence was observed from concentration of substance and from infective dose of virus. Discussion. Effective suppression of the reproduction of influenza virus strains A(H1N1), A(Н1N1)pdm09 and A(H3N2) in the different sublines of MDCK cells with «Kagocel» was shown by the different methods. These results give the possibility to suggest that along with the ability to induce interferons, «Kagocel» can impact on the reproduction of influenza virus, but the further research is needed. Conclusion. «Kagocel» substance effectively inhibits the reproduction of influenza virus strains A(H1N1), A(Н1N1)pdm09 and A(H3N2) in vitro. At the same time, the selectivity index is quite high.
Problems of Virology. 2019;64(3):125-131
125-131
THE ROLE OF THE NEONATAL FC RECEPTOR IN THE UNCOATING OF ECHOVIRUSES AND COXSACKIEVIRUS A9
Abstract
The aim of this study was to determine the role of the human neonatal receptor for the Fc fragment of IgG (hFcRn) as a common uncoating cellular receptor for echoviruses and coxsackievirus A9 during infection of human rhabdomyosarcoma (RD) cells. Material and methods. The protective effect of the human serum albumin, purified from globulins, (HSA-GF) and antibodies to hFcRn was studied in RD cells infected with several strains and clones of species B enteroviruses possessing different receptor specificity (echoviruses 3, 9, 11, 30 and coxsackieviruses A9, B4, B5). Results. It was shown that HSA-GF at concentrations of 4% or less protected RD cells from infection with echoviruses 3, 9, 11 and coxsackievirus A9. The antibodies to hFcRn at concentrations of 2.5 ug/mL or less demonstrated the similar spectrum of protective activity in RD cells against infection with echoviruses 3, 9, 11, 30 and coxsackievirus A9. The protective effect of HSA-GF or the antibodies to hFcRn was not observed in RD cells infected with coxsackieviruses B4 and B5 that need coxsackievirus-adenovirus receptor for uncoating. Discussion. The usage of the previously characterized echovirus 11 clonal variants with different receptor specificity allowed us to define the function of hFcRn as a canyon-binding uncoating receptor in RD cells. The kinetics and magnitude of the observed protective effects correlated with receptor specificity of the enteroviruses used in this work supporting the two-step interaction of DAF-dependent echoviruses with the cellular receptors. Conclusions. In this study, the function of hFcRn was defined in RD cells as a canyon-binding and uncoating receptor for echoviruses and coxsackievirus A9. The two-step interaction of DAF-dependent echoviruses during entry into the cells was confirmed: initially with the binding receptor DAF and subsequently with the uncoating receptor hFcRn.
Problems of Virology. 2019;64(3):132-139
132-139
IDENTIFICATION OF ROTAVIRUS I- AND E-GENOTYPES BY MULTIPLEX PCR METHOD
Abstract
Introduction. In recent years the presence of reassortant rotavirus strains is increasingly mentioned in the world due to the application of the full-genome based classification system. Information on the circulation of such strains in the territory of Russia is limited. The aim of this work was the development of the approach for determination of genotypes of segments encoding VP6 (I) and NSP4 (E) to reveal reassortant strains. Material and Methods. Rotavirus-positive samples were studied by means of nucleotide sequencing and multiplex PCR. Phylogenetic analysis was conducted using the Bayesian approach. Results. Three alleles of the VP6 gene (I1-1, I2-IV, I2-VII) and seven alleles of the NSP4 gene (E1-I, E1-III, E2-VI, E2-VII, E2-X, E2-XII, E3) were detected on the base of nucleotide sequences of Nizhny Novgorod rotaviruses. Taking into account these results, the oligonucleotide primers specific to genotypes I1, I2, I3 and E1, E2, E3 were designed. Optimal conditions for multiplex PCR were chosen. The method was tested using the strains collected in Nizhny Novgorod in 2018. The diversity of I and E genotypes was determined and various combinations with G and P genotypes were identified. Discussion. G9-P[8]-I1-E1 rotaviruses were predominant (32.7 %) and G2-P[4]-I2-E2 rotaviruses were in second place (29.1 %). Strains with genotypes G4-P[8]-I1-E2, G3-P[8]-I2-E2 and G2-P[4]-I2-E1 were detected sporadically. They had genes of two rotavirus genogroups, so can be considered to be reassortant. Conclusion. The proposed approach is a useful tool for the characterization of rotaviruses in the conditions of the beginning of vaccination against rotavirus infection in Russia.
Problems of Virology. 2019;64(3):140-144
140-144
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