Vol 96, No 3 (2019)

ORIGINAL RESEARCHES

Characteristics of bifidobacteria biological properties in gut microflora disorders in HIV-infected children

Zakharova Y.V., Levanova L.A., Shternis T.A., Sukhikh A.S., Marcovskaya A.A.

Abstract

Aim is to evaluate the biological properties of bifidobacteria in HIV-infected children with different degrees of gut microflora disorders to determine the direction for correction of bifidobacterium in HIV infection.

Materials and methods. 58 selected cultures of bifidobacteria from HIV-infected children, 45 children in the comparison group. In bifidobacteria studied adhesion index of microorganisms (IAM), hydrophobicity (H), acid formation, antioxidant activity (AOA), delayed antagonism to opportunistic microorganisms.

Results. In HIV-infected children at I degree of gut microflora disorders biological properties of bifidobacteria did not differ from the comparison group (p>0.05). At II degree of gut microflora disorders hydrophobicity of bifidobacteria was 45,75% (33; 58.8), IaM — 2.5 (2,0; 2.9), acid formation 67,30 T (53,0; 97,4), AOA — 0.8 Eaoa (0,6; 1.0). At III degree of gut microflora disorders H — 22,7% (15.8; 46,6), iAm — 2.4 (1,92; 3,69), acid formation — 66,3 T (48,9; 94,9), AOA-0.4 Eaoa (0,3; 0,6). Value biological properties at II and III degrees of gut microflora disorders were lower than in bifidoflora from HIV-negative children (p<0.01).

Conclusion. At the I degree of gut microflora disorders in HIV infection it is necessary to correct the quantitative level of bifidobacteria. At II and III degree it is necessary to correct the surface properties, antioxidant and fermentation of carbohydrates activity of bifidobacteria.

Journal of microbiology, epidemiology and immunobiology. 2019;96(3):3-9
pages 3-9 views

Age features of species diversity lactobacill of large intestine in residents of Tyumen

Kataeva L.V., Vakarina A.A., Kolotova O.N., Posoyuznykh O.V., Tashlanova V.V., Karpukhina N.F., Bychkova L.A.

Abstract

Aim. To study the species composition of bacteria of the genus Lactobacillus, inhabiting the distal large intestine of patients of different age groups, residents of the city of Tyumen.

Materials and methods. The content of bacteria of the genus Lactobacillus in feces of 1545 residents of Tyumen aged 1 month to 76 years who complained of intestinal dysfunction was analyzed. Grown colonies of lactobacilli identified by mass spectrometry.

Results. The study of the species composition of lactobacilli showed their wide diversity depending on age. Statistically significant differences in the content of lactobacilli in the large intestine in different age groups of patients were identified by the species L. rhamnosus and L. fermentum.

Conclusion. Information on the species characterization of lactobacilli that inhabit the large intestine of patients, taking into account age-related features, will increase the effectiveness of the correction of the microbiota of the large intestine and the prevention of inflammatory diseases of the gastrointestinal tract.

Journal of microbiology, epidemiology and immunobiology. 2019;96(3):10-15
pages 10-15 views

New forms of home dust mite allergoid

Berzhets V.M., Babakhin A.A., Petrova N.S., Vasileva A.V., Khlgatyan S.V., Emelyanova O.Y.

Abstract

Aim. The purpose of this study is to create the technology of preparation of the polymeric allergoid received by chemical modification of Dermatophagoides farinae extract by a formaldehyde and a monomeric allergoid of Dermatophagoides pteronyssinus received by succinilation and studying of their physical, chemical and immunologic properties.

Materials and methods. In the modification species determined the protein content, isoelectric points of protein components by a method of isoelectrofocusing (IEF), esterase activity. Antigenic properties of allergoid studied by methods of an immunodiffusion and an immunoelectrophoresis. Applied microdot immunoenzyme assay to characterize the specific activity of a resulting allergoid. Received by a succinilation allergoid was also studied on physical and chemical indicators.

Results. It is established That the formalinization is resulted by the expressed depression of allergenic activity of an allergoid. It is also shown that succinilation leads to essential depression of allergenicity of a monomeric allergoid due to blockade of B-cellular epitopes and to conservation of an adjuvanticity helps to T-cellular epitopes.

Conclusion. Thanks to the proved depression of allergenicity and rising of an adjuvanticity monomeric and polymeric allergoid can be recommended for carrying out allergenspecific immunotherapy (ASIT).

Journal of microbiology, epidemiology and immunobiology. 2019;96(3):15-21
pages 15-21 views

Antiretroviral therapy effect on specific antibodies spectrum changes to HIV-1 individual antigenes from persons infected with different virus subtypes

Lukhverchik L.N., Alatortseva G.I., Nesterenko L.N., Dotsenko V.V., Amiantova I.I., Zhukina M.V., Kabargina V.Y., Bobkova M.R., Kazennova E.V., Zverev V.V.

Abstract

Aim. To estimate the antibodies (Ab) level to HIV-1 antigens in persons infected with virus various subtypes, which received and didn’t receive antiretroviral therapy (ART).

Materials and methods. Blood serum samples of HIV-1-infected with subtypes A1, B and C (ART+) — 40 persons, (ART-) — 29 persons. Ab were determined by modified linear immunoassay. Positivity indices for each antigen were calculated for each sample.

Results. Multidirectional changes in the Ab level to HIV-1 antigens were revealed in persons infected with virus various subtypes at the ART background and without it.

Conclusion. The study of Ab spectrum changes in persons infected with virus various subtypes at the ART background is promising for the additional dynamic criteria development for HIV infection progress estimation.

Journal of microbiology, epidemiology and immunobiology. 2019;96(3):22-27
pages 22-27 views

Analysis of prevalence of HIV-1 primary resistance to antiretroviral drugs in the territory of Moscow and Moscow region

Nosik M.N., Ryzhov K.A., Kravchenko A.V., Sevostyanihin S.E., Kuimova U.A., Potapova А.B., Sobkin A.L.

Abstract

Aim. To analyze the level of HIV drug resistance among primary patients (who did not receive antiretroviral therapy (ART)) living in the Moscow region, where access to the antiretroviral therapy is quite high.

Material and methods. Blood samples from 527 drug-nanve patients who live in the Moscow (n=279) and in the Moscow region (n=248) were examined for the period 2008-2015 years. For detecting and analyzing possible mutations of resistance virus genome nucleotide sequence region was studied using the test systems ViroseqTMHIV-1 Genotyping System and StanfordnHIV Drug Resistance database (http://cpr.stanford.edu/cpr.cgi).

Results. It was found that the level of transmission of HIV-1 resistant strains among na^e-drug HIV-positive patients is still quite low, that is 2%. However, the percentage of polymorphism mutations and secondary substitutions which in combination with other resistance mutations in the reverse transcriptase gene can reduce the virus sensitivity to antiretroviral drugs (ARVs) is quite high — 34.5%.

Conlusion. The level of primary resistance among drug-nanve HIV-infected persons does not exceed 5%. However, given the wide access to ART and the emergence in recent years of ambiguous mutations and deletions in the reverse transcriptase/ protease gene that could potentially lead to ARVs resistance, it is obvious that continuous monitoring of the circulation of resistant strains is necessary.

Journal of microbiology, epidemiology and immunobiology. 2019;96(3):27-31
pages 27-31 views

The method to detect sIgE to thyroid peroxidase in cases of the patients with autoimmune and allergic diseases

Zubkov A.V., Kuzmina N.S., Mazurina S.A., Butova L.G., Rahmatulina N.M., Sibgatulina N.A., Gervazieva V.B.

Abstract

Aim of this research has been to create a method to detect sIgE to thyroid peroxidase (TPO) in serum of the patients with autoimmune and allergic diseases.

Materials and methods. In this research native affine-purified TPO (more the 95% clean, enzyme activity 60±15 of enzyme unit per mg) with immune dominant elements only (confirmed by inhibitory ELISA) has been used. The detection of sIgE in serum, combined with TPO, was carried out with monoclonal antibodies to IgE (8E/4F4) conjugated with horseradish peroxidase.

Results. In serum of patients with bronchial asthma, allergic rhinitis and chronic spontaneous urticarial sIgE to TPO has been detected in 18—21% of cases. The healthy people and the patients with autoimmune thyroiditis had no sign of sIgE to TPO in their serum.

Conclusion. The detection of sIgE to TPO in cases of patients with chronic spontaneous urticarial and allergic diseases will make possible to perform the early diagnosis of thyroid autoimmune disorders and provide timely and adequate therapy.

Journal of microbiology, epidemiology and immunobiology. 2019;96(3):31-36
pages 31-36 views

Specific humoral immunity after immunization with vaccine contained Leningrad-3 (L-3) mumps virus strain of different potency

Оtmshevskaya E.V., Kulak M.V., Bukin E.K., Ignatev G.M.

Abstract

Aim. The persistence of the specific antibodies and the kinetics of the antibody neutralizing activity level and spectrum were studied in two groups of healthy young adults after the immunization with two different vaccine lots characterized with the different vaccine potency were studied for 3 years.

Materials and methods. Antibodies were measured by enzyme immunoassay (EIA) and by plaque reduction neutralization (PRN) assay using the L-3 vaccine mumps virus (MuV) and five more geterologous MuV strains of genotypes A, B, C, D and H as the targets in 3-year follow-up serum samples (n=60) of once-vaccinated volunteers.

Results. Maximal of the specific antibodies functional activity according to the avidity index as well as to the neutralizing activity level against 6 used in the investigation MuV strains was registered on month 12th till 18th after vaccination in both groups of volunteers. Interestingly the specific IgG levels registered by EIA were going already down from month 12th. It was a waning immunity tendency, in general, over time in both groups of volunteers. In the current investigation two vaccines lots contained significantly different MuV titer in one dose (2, 76-fold difference, U-test p>0,005) induced the specific humoral immunity with no statistical differences in the measured parameters in 60 once vaccinated volunteers.

Conclusion. The current study results make it possible to suggest controlling or limiting the maximum permissible level of vaccine potency, as no more. Such limitation could reduce the risk of the post vaccination reaction and of vaccine virus horizontal transmission. At the same time the optimal vaccine potency bound will make vaccine production more economical.

Journal of microbiology, epidemiology and immunobiology. 2019;96(3):37-45
pages 37-45 views

Interferon status in estimating the therapy of bronchial asthma with immunomodulating preparations

Ospelnikova T.P., Zarembo N.V., Konischeva A.Y., Gervazieva V.B., Osipova G.L., Michailova N.A.

Abstract

Aim: to investigate the violations of the interferon status and to evaluate the clinical and immunological efficacy of immunoactive drugs in patients with asthma with the justification of their use in persons with an immunocompromised viral background.

Materials and methods. We examined 86 patients with atopic asthma, 67 patients with atopic BA of the lung, medium and moderate course during remission of the disease, and 19 patients with asthma exacerbation. Immunovac VP-4 was used in patients with BA in remission, interferon inducer (IFN) Cycloferon in patients with exacerbation of BA. The observation period of outpatients was 1 year, during which they monitored clinical, allergological and immunological parameters, including the assessment of interferon status with the determination of the biological activity of IFN type I and II, as well as the activity and quantitative content of type II IFN and IL-4 in the blood serum.

Results. All patients with asthma, regardless of the severity of the disease, showed a pronounced decrease in the biological activity of IFN types I and II produced by the patients’ leukocytes, and a reduced potential of Th2 cells to synthesize IL-4 during remission, incomplete remission, and especially in period of exacerbation caused by ARVI.

Conclusion. The inclusion of immunomodulating drugs Immunovac VP-4 and cycloferon in the complex therapy of patients with atopic asthma of light and moderate severity leads to an increase in leukocyte IFN-producing ability, a decrease in the incidence of ARVI and the number of exacerbations of the underlying disease. The use of synthetic and bacterial immunomodulators in patients with immunocompromised genesis of the disease will reduce the number of exacerbations of asthma and the frequency of respiratory viral infections.


Journal of microbiology, epidemiology and immunobiology. 2019;96(3):46-54
pages 46-54 views

The application of molecular genetic analysis and genomic profiling of infectious diseases causative agents in Sochi during the preparation and holding of the Fifa World Cup 2018

Popova A.Y., Kulichenko A.N., Volynkina A.S., Kuznetsova I.V., Podkolzin A.T., Chekhvalova E.V., Orobey V.G.

Abstract

During the preparation of the FIFA World Cup 2018 in 2015-2017 genetic features of regional pathogenic strains in the city of Sochi are determined. The strains and nucleic acid isolates of bacterial and viral infectious diseases causative agents — acute intestinal infections: rota-, noro-, astro and enteroviruses, Salmonella enteritidis, natural focal infections: hantaviruses, Rickettsia of tick spotted fevers group, Borrelia are genotyped. In result, complex genetic profiling of infection diseases causative agents variants that are actual for the Sochi region has been carried out. The obtained database was used to provide molecular analysis of pathogens during the XIX International Festival of Youth and Students (2017) and during the FIFA World Cup (2018). The perspective of using this algorithm of genetic analysis in description of outbreaks (cases) of infectious diseases in specific areas is shown.

Journal of microbiology, epidemiology and immunobiology. 2019;96(3):54-59
pages 54-59 views

Pre-clinical study of protective potency of candidate recombinant live pertussis vaccine for intranasal administration

Sinyashina L.N., Semin E.G., Medkova A.Y., Syundyukova R.A., Karataev G.I.

Abstract

The aim of present research is studying of protective potency of candidate recombinant live pertussis vaccine (RLPV) for intranasal administration.

Materials and methods. Two methods of protective potency assessment, based on mice survival estimating, were used: after intracerebral administration of B.pertussis 18323 bacteria and after intranasal administration of virulent Bordetella spp. bacteria (B.pertussis, B.parapertussis and B.bronchiseptica) to immunized mice.

Results. An ingenious method of pertussis vaccine protective potency assessment due to mucosal immunity is suggested. Protective potency of constructed RLPV estimated in two tests of intracerebral and intranasal administration was higher than market image drug of DTP vaccine. RLPV provided protection from infection in mice after administration of both wild type and recombinant Bordetella spp. bacteria.

Conclusion. Examined recombinant live pertussis vaccine (RLPV) for intranasal administration has significant protective potential and could be recommended for using in clinical trials.

Journal of microbiology, epidemiology and immunobiology. 2019;96(3):60-69
pages 60-69 views

Efficiency of combined therapy of figuratum and parainfectious erythemas with the use of immunomodulators of different origins and MNRI

Sorokina E.V., Akhmatova N.K., Egorova N.B.

Abstract

The aim of the study was to study the etiopathogenesis of erythema, comparative study of the clinical efficacy of combination therapy with the use of immunomodulators in erythema, the study of the dynamics of immunological parameters as a result of therapy.

Materials and methods. The study included 215 adult patients with erythema. Before treatment and 1.5 months after therapy in patients have been studied: the expression of TLRs on MLPC and skin cells by flow cytometry with the use of the mat for TLR2, TLR3, TLR4, TLR5, TLR6, TLR7, TLR8, TLR9 (Caltag Laboratories, USA) using a flow cytometer FC-500 (Beckman Coulter, USA); the subpopulation composition of peripheral blood lymphocytes; the levels of pro-inflammatory and regulatory cytokines in the serum by the method of solid-phase ELISA using test systems company «Biosource» (Austria); the main classes of immunoglobulins (IgG, IgM, IgA) in serum.

Results. It was shown that these pathogenetic and diagnostic significance of viruses of the family nerpesviri-dae, bacterial contamination by different types of microorganisms (Staphylococcus aureus, Klebsiella pneumoniae, Streptococcus spp., Proteus vulgaris). A substance containing antigens of conditionally pathogenic microorganisms, and the use of neuro sensorimotor reflex integration (MNRI) contributed to the decline in the number and severity of recurrences of erythema, the extension of the range expressed Toll-like receptors; increase of lymphocytes with immunophenotype CD3+, CD4+, CD25+, CD95+; increase in IFN-y, decreased IL-4; largely reduced the level of total IgE than other types of therapy.

Conclusion. The inclusion in therapy of patients with erythema, immunomodulator of bacterial origin and MNRI promotes increase of clinical efficiency and correlates with the correction of immunological disorders.

Journal of microbiology, epidemiology and immunobiology. 2019;96(3):69-76
pages 69-76 views

Mixed infectious disease forecasting technique, based upon seasonal decomposition and Sarima

Filatova E.N., Solntsev L.A.

Abstract

Aim. To study the possibility of using mixed technique for predicting infectious morbidity based on time series decomposition methods and SARIMA (decSARIMA).

Materials and methods. Using the data from 12 regions of Volga Federal District (Russia) we analyzed time series of the incidence of infectious pathologies: hemorrhagic fever with renal syndrome (HFRS), acute upper respiratory viral infection (ARVI) and syphilis. The decomposition of time series of the incidence rate was carried out using X13-ARIMA-SEATS method. The trend and the seasonal component were separated, each of which was then modeled separately by SARIMA method. The final model of the incidence rate was obtained by adding the trend and the seasonal models.

Results. On average, decSARIMA models had higher or similar characteristics of model and prediction quality compared to SARIMA models without preliminary decomposition. The prognosis of the incidence rate obtained by decSARIMA method was characterized by narrower confidence intervals. Reasonability of using decSARIMA models depended on composition and dynamics of time series of the incidence rate. A significant improvement in model and prediction quality was demonstrated for HFRS. When modeling and predicting the incidence rate of ARVI and syphilis, the inclusion of decomposition of time series into the analysis was considered inexpedient.

Conclusion. The usage of decSARIMA model allows to significantly improve the quality of the prognosis of the incidence for infections, which are characterized by pronounced seasonality and the presence of interannual differences in the incidence rate.

Journal of microbiology, epidemiology and immunobiology. 2019;96(3):76-83
pages 76-83 views

REVIEWS

Biohazard caused by viable but nonculturable microorganisms

Pakhomov Y.D., Blinkova L.P.

Abstract

In this review we discuss current notions about viable but nonculturable microorganisms — new subject of microbiological research. Information about factors and conditions of formation of nonculturable cells and their resuscitation is analyzed. Of particular interest is information of biohazard of nonculturable cells that are hard to isolate by traditional means from organisms, the environment, foodstuffs etc. Knowledge about this is valuable for detection live microbes from clinical samples, levels of viable but nonculturable cells in vaccine and probiotic preparations, culture collections.

Journal of microbiology, epidemiology and immunobiology. 2019;96(3):83-91
pages 83-91 views

Interferon status as a method of determination of nonspecific biomarkers of human immunopathology

Ershov F.I., Ospelnikova T.P., Narovlyansky A.N.

Abstract

For more than 60 years, the study of interferons (IFN) system has continued, as a complex network system. IFNs of 3 types are discovered: I (α/β), II (ƴ), III (λ), their interrelations, mechanisms of action, functional diversity are investigated. The practical way out of the study of the functional capacity of human peripheral blood leukocytes to produce IFN was the method of determining the interferon status, which allows to judge the immunoreactivity of the organism, to detect the sensitivity of blood cells to immunoac-tive drugs and makes it possible to determine the tactics of treatment in different forms of pathology and predict the outcome of the disease. An improved method of IFN status has been proposed and scientifically justified, the indicators of which can now be considered as characteristics of nonspecific biomarkers of human immunopathology.

Journal of microbiology, epidemiology and immunobiology. 2019;96(3):91-99
pages 91-99 views

Yersinia pestis pathogenicity

Tynyanova V.I., Sokolova E.P., Zyuzina V.P., Demidova G.V.

Abstract

Yersinia pestis belongs to those pathogenic bacteria which produce lipopolysaccharide (LPS) having the function of a toxin. In order to make a toxic effect the polymer must be separated from the cell outer membrane and presented to the immunocompetent cell receptors of the host in the functionally active form. In this review data of russian and foreign investigators on Y. pestis toxigenic properties was presented. Results of the authors' own experiments showing that Y. pestis is able to export LPS into the surrounding medium are included. This process is a natural function of the living cell, is realized at 37 degrees C and is strictly dependent on the expression of Y. pestis genes of extrachromosomal inheritance, pMT1, pCD1, pPCPl. By the use of isogenic variants of Y. pestis EV76 vaccine strain and virulent 231 strain containing different plasmid combinations, it was established that maximum contribution in the activation of «high-temperature» LPS and its transformation into extracellular form made the proteins encoded by pCD1. The significance of the «murine» toxin encoded by pMT1 plasmid was less pronounced. The participation of pPCPl plasmid in the toxic effect was not discovered. The role of Y. pestis capsular substance and the significance of biologically active factors in the realization of Y. pestis LPS toxic potential is discussed. Functional relationship between translocation of the proteins encoded by plasmids and Y. pestis toxigenicity suggests Y. pestis biological uniqueness.

Journal of microbiology, epidemiology and immunobiology. 2019;96(3):99-109
pages 99-109 views

Method of Immuno-PCR in diagnostics of bacterial and viral infections

Barkova I.A., Barkov A.M., Viktorov D.V.

Abstract

Immuno-PCR (I-PCR) combines the capabilities of two modern diagnostic methods of enzyme immunoassay (ELISA) and polymerase chain reaction (PCR), combination these methods causes a 100-10000 fold increase in sensitivity compared to a similar ELISA. The review considers the main variants of I-PCR, gives examples of possible use of the method for early and retrospective diagnosis of various diseases.

Journal of microbiology, epidemiology and immunobiology. 2019;96(3):110-117
pages 110-117 views

The enzyme-linked immunosorbent assay (ELISA): history, theory and application

Tarakanova Y.N., Dmitriev A.D., Dmitriev D.A., Lavrov V.F., Massino Y.S., Pechelyulko А.A., Segal O.L.

Abstract

Enzyme-linked immunosorbent assay (ELISA) play a significant role in the development of many important fields of biology and medicine, including the detection of the dangerous infections agents. In the present review there is described the history of ELISA’s invention, further improvement of this method and application in modern biology and medicine. The special accent is made on factors influencing antibody-antigen interactions on the solid phase and their use to increase the analytical sensitivity of the method.

Journal of microbiology, epidemiology and immunobiology. 2019;96(3):117-125
pages 117-125 views


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