Vol 5, No 4 (2015)

REVIEW ARTICLES

MOLECULAR-EPIDEMIOLOGICAL FEATURES OF HEPATITIS B VIRUS

Gerasimova V.V., Levakova I.A., Bichurina M.A., Maksimova N.R.

Abstract

Abstract. Hepatitis B virus is a serious issue of public health services all over the world, particularly in Russia and its regions. The cause can be determined due to its higher incidence rate in different ethnic and geographical groups, variety of manifestation of clinical forms as well as outcomes. According to WHO data, about 50 million people in the world are annually infected with hepatitis B virus. Development of molecular-genetic methods of research has allowed to expand representations about the activator biology. Genetic variability of hepatitis B virus genome has been subject to identification of 10 various geographically widespread genotypes designated by letters from A to J. In a number of genotypes subgenotypes differing from one another on 4–8% full genome sequences are revealed. In the work a table of geographical prevalence of various genotypes of hepatitis B virus in the world is presented. The geographic distribution of genotypes of hepatitis B virus is closely connected with endemic regions and indigenous population, living there. So the genotypes B and C are connected with the population of Asian countries, genotypes A and D are extended among European countries and in the USA. The genotype D is considered to be a prevailing type among patients with hepatitis B in several regions of Turkey. The phylogenetic analysis of nucleotide sequences of hepatitis B virus isolates in African regions has shown presence of three subgenotypes — А1, А2, А3. In the countries of Asia the following six subgenotypes B are noted: В1 in Japan, В2–В5 and В7 in East Asia Subgenotip B6 is found among indigenous population living in the Arctic regions, including Alaska, northern Canada and Greenland. The genotype C has five serotypes (С1–С5) which are extended in East and South East Asia. The genotype D, also has five subgenotypes (D1–D5) which are extended in Africa, India, the Mediterranean region, Europe. Genotype F has four subgenotypes (F1– F4) which circulate in Central and the South America. Subgenotypes are not revealed among genotypes E, G, H, I and J. In the territory of the Russian Federation the highest incidence of hepatitis B virus is noted in 3 genotypes (A, C, D). Among them the genotype D comprising 88% of them is the prevailing one. 

Russian Journal of Infection and Immunity. 2015;5(4):297-302
pages 297-302 views

Rgg-LIKE TRANSCRIPTIONAL REGULATORS IN STREPTOCOCCUS spp.

Dmitriev A.V., Chaussee M.S., Kalinina O.V.

Abstract

An ability of pathogenic bacteria to survive in different ecological niches, to successfully adapt to changing environments, to colonize different organs and tissues, and to cause numerous diseases in human and animals including severe invasive diseases is provided, in particular, by the presence of specific proteins involved in regulation of gene transcription. This review summarizes the current data on the Rgg-family (TIGR01716 family, The Institute for Genomic Research, http://www.jcvi.org) of regulatory proteins encoded by some of the low G+C gram positive bacteria such as Streptococcus gordonii, Streptococcus agalactiae, Streptococcus pneumoniae, Streptococcus mutans, Streptococcus thermophilus, Streptococcus suis and Streptococcus pyogenes. Proteins of this family has helix-turn-helix (HTH) motif at N-terminus which is able to bind promoter regions of the genes and regulate their transcription. The mechanisms of Rgg-dependent transcriptional regulation and the role of certain amino acids for functioning of Rgg-like proteins are discussed. The Rgg-like regulators have evolved to regulate diverse set of genes associated with virulence, metabolism, stress response, competence, biofilm formation, etc. The Rgg-like regulators are also involved in quorum sensing. Rgg-like proteins regulate not only the genes located adjacently to rgg, but also distantly located genes. Rgg-like proteins of different bacterial species have certain sequence similarity, and it is suggested that their genes are horizontally acquired. Rgg-dependent transcriptional regulation varies in a strainand species-specific manner that supports the hypothesis of the complexity of transcriptional regulation in gram-positive bacteria. The current review also discusses the role or Rgg-like regulators in control of virulent properties of gram-positive bacteria and their interaction with human host. Given the importance of Rgg-like regulators for virulence, these proteins (their genes or transcripts) can be considered as targets for development of the novel selective agents against different bacterial infections. 

Russian Journal of Infection and Immunity. 2015;5(4):303-314
pages 303-314 views

ORIGINAL ARTICLES

PRODUCTION OF MYCOBACTERIUM TUBERCULOSIS TB10.4 RECOMBINANT PROTEIN IN ESCHERICHIA COLI

Dukhovlinov I.V., Fedorova E.A., Dobrovolskaya O.A., Bogomolova E.G., Chernyaeva E.N., Al-Shekhadat R.I., Simbirtsev A.S.

Abstract

Nowadays tuberculosis is considered one of the most dangerous infectious diseases occurring everywhere, and it remains a cause of death of millions of people around the world. According to the World Health Organization data, in 2013 tuberculosis caused more than 9 million cases worldwide and about 1.5 million of infected people died. The causative agent of tuberculosis in most cases is Mycobacterium tuberculosis. But sometimes it can be Mycobacterium bovis or Mycobacterium africanum. Mainly as a result of infection, a bacterial infection affects the lungs, but the disease may develop in other organs and tissues. Now for the prevention of tuberculosis vaccination of newborns with attenuated vaccine BCG is widely used. The production of this vaccine is cheap and it is safe to use. Thus today, vaccination is the primary means of prevention of tuberculosis. However dubious efficacy and a number of side effects observed after vaccination, makes the scientific community to develop new effective methods for the treatment of tuberculosis. One of the ways to develop new vaccines against tuberculosis is to provide a subunit vaccine based on recombinant proteins. Advantages of subunit vaccines are that the preparation containing the purified protein is stable and secure, its chemical properties are known, it does not contain additional proteins and nucleic acids, which could cause undesirable effects in the human body. One of the most promising antigens for use as components in new vaccines is considered a low molecular weight secreted protein TB10.4. TB10.4 protein is recognized at an early stage of tuberculous infection and contributes to the proliferation of lymphocytes responsible for the production of IFNγ. TB10.4 protein also possesses an adjuvant effect when administered in combination with mycobacterial proteins. Given these properties, the recombinant protein TB10.4 can be used to generate new candidate vaccines against tuberculosis. During the study was created high-yield E. coli strain, which produces the recombinant protein TB10.4, selected the optimal protocol of induction of the gene encoding the protein. The protein was purified using metal affinity chromatography. The purity of the final preparation reached 98%. 

Russian Journal of Infection and Immunity. 2015;5(4):315-322
pages 315-322 views

THE INVARIANT PATTERNS OF THE INTERNAL PROTEINS OF PANDEMIC INFLUENZA VIRUSES

Kharchenko E.P.

Abstract

The purpose of the study was to find molecular recognition characteristics of pandemic strains of influenza A viruses and to find out whether avian strains are the probable cause of a new influenza pandemic. Computer analysis of the internal proteins (nucleoprotein, matrix protein M1 and M2 proteins polymerase complex PB1, PB2 and PA, non-structural protein NS2; because of the variability of the length the non-structural NS1 protein was excluded from the analysis) of influenza A virus pandemics in 1918 (H1N1), 1957 (H2N2), 1968 (H3N2), 1977 (H1N1) and 2009 (H1N1) strains was used for search of the invariant pattern primary structure. It was revealed that internal proteins of pandemic strains are characterized by the constancy of the number and positions of certain amino acids and the presence of extended invariant fragments. On the basis of these identified patterns of invariances in internal proteins it was possible to accurately identify pandemic strains in the control sample. Pandemic strains, divided by decades in their emergence and different composition of subtypes of hemagglutinin and neuraminidase (H1, H2, H3 and N1, N2), have strong relationship for their internal proteins, forming a special subset. This suggests that emergence of influenza A virus pandemic strains is related to convergence of their internal proteins to the detected pandemic invariants. To identify pandemic invariant patterns is enough to have the training set including strains of four pandemics (1918, 1957, 1968, 1977). Therefore the 2009–2010 pandemic influenza strain could be predicted at the earliest stage according to its genome and proteome sequencing. According to a comparative analysis, the internal proteins of avian strains H5N1 and H7N9, particularly their nucleoproteins, are not close to those of pandemic strains. This suggests that the threat of a new influenza pandemic, provoked by current circulating avian strains, is unlikely. Invariant patterns of pandemic strains can potentially be used to track pre-pandemic strains among circulating influenza A viruses and detect the formation of a possible trajectory of pandemic alert. 

Russian Journal of Infection and Immunity. 2015;5(4):323-330
pages 323-330 views

ASSOCIATION OF TREM-1 GENE POLYMORPHISMS WITH INFECTIVE ENDOCARDITIS

Ponasenko A.V., Kutikhin A.G., Khutornaya M.V., Yuzhalin A.E., Rutkovskaya N.V., Golovkin A.S.

Abstract

Infective endocarditis (IE) is a septic inflammation of endocardium, which generally involves the lining of the heart chambers and heart valves. The development of IE depends in many respects on how properly and efficiently the immune system responds to the occurrence of an infection. Innate immunity, which carries out the response to a transient bacteremia, is genetically determined in a large extent. Pattern recognition receptors, which identify pathogenand danger-associated molecular patterns, are the main effectors of innate immune response; one of these receptors is triggering receptor expressed on myeloid cells-1 (TREM-1). We hypothesized that inherited variation in TREM-1 gene may affect individual susceptibility to IE. The distribution of genotypes and alleles of rs1817537, rs3804277, rs6910730, rs7768162, rs2234246, rs4711668, rs9471535, and rs2234237 gene polymorphisms was investigated in 110 Caucasian (Russian) subjects with IE and 300 age-, sex-, and ethnicity-matched healthy blood donors. Odds ratios with 95% confidence intervals were calculated. We found that rs1817537 polymorphism was associated with decreased IE risk (OR = 0.60; 95%CI = 0.37–0.99; р = 0.046, dominant model); however, this was not significant after an adjustment for multiple comparisons. Therefore, we observed no statistically significant association between the investigated polymorphisms within TREM-1 gene and IE. Further in-depth investigations in this field are necessary to shed the light on the impact of inherited variation within innate immune response genes on the development of IE. 

Russian Journal of Infection and Immunity. 2015;5(4):331-338
pages 331-338 views

BLOOD LEUKOCYTES PHENOTYPING BY HEMATOFLOW METHOD IN PATIENTS WITH ONYCHOMYCOSIS

Savchenko A.A., Borisov A.G., Anisimova E.N., Belenyuk V.D., Kudryavtsev I.V., Reshetnikov I.V., Kvjatkovskaja S.V., Cejlikman V.J., Zorin A.N.

Abstract

The aim of the investigation was to evaluate the information content of Hematoflow method in the pathogenetic significance in determining the violations of the cellular responses of innate and adaptive immunity, as well as the decision on the appointment of immunotropic treatment of patients with onychomycosis. The study involved 42 patients with onychomycosis feet and hands/feet at the age of 20–45 years before the appointment of a systemic antifungal therapy. The diagnosis of mycosis, onychomycosis was confirmed by microscopic examination of the fragments of the damaged nail plate. The growth of the fungus culture on special media was observed in 64% of patients. 24 healthy persons were examined as controls. A study of the phenotype of white blood cells was performed on a dual-platform technology hematology analyzer and flow cytometry using a set of antibodies Cytodiff: CD36-FITC, CD2-PE, CD294(CRTH2)-PE, CD19-ECD, CD16-PC5 и CD45-PC7. The phenotypic composition evaluation of the white blood cells by the Hematoflow method allowed to establish in patients with onychomycosis the violation cellular innate and adaptive immunity. Minor changes were detected in the population composition of granule cells in the peripheral blood of patients manifested to an increase in the content of the young and segmented granulocytes. When monotsitopeniya patients with onychomycosis increases the content of the «classic» monocytes and decreases the level of «non-classical» monocytes. Changes in the composition of blood monocytes subpopulation identified in patients with the infection lasting up to 3 years and stored in the course of the disease. The most pronounced changes were found in patients with onychomycosis by the performance of adaptive immunity. Lymphopenia in these patients is realized by reducing the number of immature and mature B-cell, but by increasing the content of T-lymphocytes. Moreover, if the content of immature B-cells have decreased in patients with a duration of infection of up to 3 years, the change in the number of mature Tand B-lymphocytes detected during disease duration 3–10 and 10 years. These changes in the content of Tand B-lymphocytes reflect immunopathogenetic processes and determine the importance of Tand B-cell immunity in onychomycosis. In general, the Hematoflow method is informative in assessing violations the cell of innate and adaptive immunity. It allows to evaluate the severity of the immunopathological process mechanism and the level of damage to the immune system, can recommend its use for a personalized approach to the appointment immunotropic treatment. 

Russian Journal of Infection and Immunity. 2015;5(4):339-348
pages 339-348 views

CYTOKINES AND HERPESVIRUSES IN CHILDREN WITH MULTIPLE SCLEROSIS

Zheleznikova G.F., Skripchenko N.V., Ivanova G.P., Surovtseva A.V., Monakhova N.E.

Abstract

It was determined earlier (G.P. Ivanova, 2012) that a chronic course of leukoencephalitis in teenagers caused by inadequate response of cytokine system to the combination of two herpesviruses (HV) — EBV and HHV-6, leads to the development of multiple sclerosis (MS) in 44% of cases. The research objective was to characterize the cytokine response in children with MS with simultaneous screening of the presence of active HV infections. 39 children with the diagnosis “MS” were under observation, 34 of them had relapsing-remitting (RR) MS, and 5 children had a progressing course of MS (PMS). Concentration of cytokines IL-1β, IL-6, IL-8, IL-10, IFNα, IFNγ, and IL-4 was identified in blood serum and cerebrospinal liquid (CSF) by enzyme-linked immunosorbent assay, HV DNA was revealed by PCR. Cytokine status in children with MS had some differences depending on the phase of the disease, clinical severity of the relapse and the course of MS. The relapse phase of RRMS was associated with the accumulation of IL-8, IL-10, and IL-6 in the blood, and index IFNγ/IL-4 modulations in accordance with the clinical severity of the relapse. A severe aggravation of the disease in children with PMS was accompanied by the increase of IL-8 system response. HV DNA was revealed in 27 patients from 39 ones (69%) in blood and in 17 patients (44%) in CSF with the predominance of EBV (93%), frequently in combination with HHV-6. During an acute period the frequency of HV DNA identification increased 2–3 times to compare with the remission period. Unlike children with RRMS, a mixed-infection of 3–4 herpes viruses was revealed in all 5 patients with PMS. According to the results summary it is possible to make a conclusion that HV-infection has an important role in MS pathogenesis in teenagers, taking part in the aggravation and progression of the disease by its effect on the cytokine system response. EBV-infection dominates among HV, however the risk of MS development increases to a great extent in the case of EBV association with another lymphotropic virus — HHV-6. 

Russian Journal of Infection and Immunity. 2015;5(4):349-358
pages 349-358 views

MOLECULAR EPIDEMIOLOGY FEATURES OF HEPATITIS C VIRUS ISOLATES FROM DIFFERENT REGIONS OF THE REPUBLIC SAKHA (YAKUTIA)

Semenov A.V., Ostankova J.V., Gerasimova V.V., Bichurina M.A., Kozlov A.V., Mukomolov S.L., Totolian A.A.

Abstract

According to WHO data about 3% of population are infected by hepatitic C virus (HCV) worldwide. Chronic hepatitis C is the leading cause of cirrhosis and hepatocellular carcinoma, thus it becoming one of the global public health problems. Clinical manifestations are varied and depend mainly of the virus biological properties and its interaction with the host immune system. Determination of virus genotype and subtype is important for a better understanding of the epidemiological and virological features of the disease. The prevalence genotypes hepatitis C virus is varies in different geographical regions of the world. The data about HCV genotypes distribution in some Russian Federation regions are very limited, especially about HCV genotypes prevalence in Siberia, Far East and some rural regions. One of such regions is Yakutia. In our study we identified genetic variants of HCV in chronic hepatitis C patients with moderate and high viral load from Yakutia by direct sequencing of HCV RNA NS5B region. Based on phylogenetic analysis we found the prevalent genotype 1 (88.3%), than genotype 2 (6.7%) and 3 (3.2%) among HCV patients with moderate and high viral load. Our results on the prevalence of subtype 1b are consistent with the data on the connection between this subtype with high levels of viremia, greater duration and severity of liver disease, as well as the development of chronic hepatits C in patients infected by HCV subtype 1b, compared with those infected with other subtypes of hepatitis virus C. The similarity of some Yakutian isolates with isolates from the United States, Brazil and Ireland was found. We discuss HCV subtype 2a isolates identified origin from isolates found in China. First in the territory of the Russian Federation HCV subtype 3g was identified, presumably imported from South Asia. Interconnected use of molecular, virological, demographic and epidemiological methods and information to monitor the infections will contribute to the understanding of the current HCV epidemiology in Russia. 

Russian Journal of Infection and Immunity. 2015;5(4):359-372
pages 359-372 views

SHORT COMMUNICATIONS

SELECTION OF PHYLOGENETICALLY CLOSELY-RELATED YERSINIA PESTIS STRAINS DIFFERING IN THEIR VIRULENCE FOR GUINEA PIGS

Anisimov N.V., Kombarova T.I., Platonov M.E., Ivanov S.A., Sukhova M.A., Dentovskaya S.V., Anisimov A.P.

Abstract

Genomic, transcriptome or (and) proteomic comparison of closely related virulent and avirulent microbial strains underlies the search for new pathogenicity factors, potential molecular targets for etiotropic therapy, vaccine prevention and immunotherapy of infectious diseases. This investigation was aimed in testing the ability of method of testicular animalization to select phylogenetically close pairs of Y. pestis strains, which dramatically differ in their pathogenicity for guinea pigs, from the populations of as a rule subcutaneously avirulent for guinea pigs “vole” strains of the plague pathogen. Animalization of Y. pestis cultures were performed on guinea pig males by fourfold testicular passage with reducing infective dose. There was no correlation between the ability to cause generalized infectious process (death) after testicular and subcutaneous infection of guinea pigs, but testicular passages made it possible to enrich bacterial culture with a portion of microbes displaying high virulence after subcutaneous infection of this animal species. The methodical approach under study can be successfully applied for selection of pairs of phylogenetically closely related bacterial strains, dramatically differing in their degrees of selective virulence. 

Russian Journal of Infection and Immunity. 2015;5(4):373-376
pages 373-376 views

THE AGGREGATION OF BACTERIA KLEBSIELLA OXYTOCA AND KLEBSIELLA PNEUMONIAE UNDER THE INFLUENCE OF CHEMICAL FACTOR

Sadrtdinova G.R.

Abstract

The article acknowledges the formation of bacterial biofilms in strains of bacteria species Klebsiella oxytoca and Klebsiella pneumoniae when grown in liquid media under the influence of negative factors (chemical factor) containing agents. Biofilms, as a community of microorganisms cause many chronic infections (meningitis, inflammatory diseases of the oral cavity, urogenital infections) and create problems in the industry (fouling of processing equipment, ship hulls, oil platforms, biocorrosion metal products). Ordinary disinfectants, such as chlorine and sodium chlorite, can not remove the biofilm, so finding an effective means of dealing with them is enough actual problem. Various antibacterial agents are ineffective in combating biofilms, since bacteria produce large amounts of polysaccharides substances that help the colony stay without disintegration. Polysaccharide serves as a barrier layer for substances in water, including for biocides. This is the main reason for the survival of microorganisms even in the heavily chlorinated water. In the study the latest data took into account on the subject, especially concerning adverse effects of oxygen on the growth of bacterial cells and directs action as a factor in the formation of biofilms. In our study we analyzed the latest generation disinfectant as an influencing factor. Working concentrations were shown in three embodiments. The number of strains studied was 6 (3 strains of each species). All strains were obtained from the Department of Museum MVE and VSE Ulyanovsk State Agricultural Academy n.a. P.A. Stolypin. In our research the biofilm community formation phenomenon has been confirmed, marked differences in biofilm formation, depending on the intensity (in this case, concentration) of the promoter and bacteria species. In vivo biofilm is easily destroyed by mechanical action (shaking test tubes with the medium). Biofilm recovery after this manipulation was not observed. The results of further studies on solid medium (meatpeptone agar) and the color smear simple dye (Gram) into the death of the bacterial cells have not been confirmed. In all cases, bacterial growth was observed, characteristic of these types of bacteria. The studies were conducted in 2015 on the basis of the Research Innovation Center of Microbiology and Biotechnology (Ulyanovsk) and with the financial support of the Foundation for Assistance to Small Innovative Enterprises in the scientific and technical sphere. 

Russian Journal of Infection and Immunity. 2015;5(4):377-381
pages 377-381 views

OBITUARY

IN MEMORY OF OLEG I. KISELEV

Abstract

IN MEMORY OF OLEG I. KISELEV 

Russian Journal of Infection and Immunity. 2015;5(4):382-382
pages 382-382 views

Instructions to authors

Abstract

Instructions to Authors 

Russian Journal of Infection and Immunity. 2015;5(4):383-385
pages 383-385 views

Author index. Subject index

Abstract

Author index. Subject index
Russian Journal of Infection and Immunity. 2015;5(4):386-386
pages 386-386 views


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