Vol 8, No 3 (2018)

The aim of the present systematic literature review is to summarize data on the role of TLRs in maintaining homeostasis of the female genitals, in maintaining the physiological development of pregnancy, provision of anti-infective resistance in pregnant women with intrauterine infection. The review substantiates the importance of TLRs of female genitals as a necessary and determining factor in the reaction to various changes in the environment, and also responsible for changes in metabolic, structural, or energy, in the maintenance of anti-infective resistance and homeostasis. As universal regulators of vital activity of organism TLRs in conjunction with other receptors of innate immunity provide maintaining the general reactivity and anti-infective resistance at the physiological level. In physiologically developing pregnancy in a background of immunosuppression in response to pregnancy TLRs during contact with infectious and non-infectious pathogens stimulate the production of nonspecific adaptive immunity factors (defensins, cathelicidins, histatines, etc.), which together with the non-specific innate factors lysozyme, complement, properdin, etc. support antiinfective resistance of the female genitals at a high level at the beginning of the infectious process. Possible violations of the development of pregnancy may be accompanied by changes in the response of TLRs to infectious and non-infectious factors until hyper-reaction, excessive inflammation or apoptosis, which requires adequate management of pregnancy. Was established the significance of the influence of pathogens of infectious and noninfectious origin in intrauterine infection indirectly through TLRs in the homeostasis of the organism, on the formation of breaches in anti-infective resistance at the organism and community level the identification of new pathophysiological and immunological pathogenetic mechanisms of development of pathological processes. IUI is a penetration of microorganisms into the tissues of fetus and it’s infection. The inhibition of the functional activity of TLRs is accompanied by the direct effect of the pathogen on the tissues, and during hyper-reaction of TLRs to pathogens revealed a pronounced inflammatory response in the fetus. The level of expression of TLRs correlates directly with the severity of the process that can be considered as early markers of infection. Depending on the nature of the pathogen an increased expression of one or the other TLRs is observed. Explained the lack of symptoms, the possibility of atypical manifestations, the asymptomatic course of infection.

Microfold cells (M cells) are specialized intestinal epithelial cells that initiate mucosal immune responses. These unique phagocytic epithelial cells are specialized for the transfer of a broad range of particulate antigens and microorganisms across the follicle-associated epithelium (FAE) into the gut-associated lymphoid tissue (GALT) by a process termed transcytosis. The molecular basis of antigen uptake by M cells has been gradually identified in the last decade. Active sampling of intestinal antigen initiates regulated immune responses that ensure intestinal homeostasis. The delivery of luminal substances across the intestinal epithelium to the immune system is a critical event in immune surveillance resulting in tolerance to dietary antigens and immunity to pathogens (e.g., bacteria, viruses, and parasites) and their toxins. Several specialized mechanisms transport luminal antigen across the gut epithelium. Discovery of M cell-specific receptors are of great interest, which could act as molecular tags for targeted delivery oral vaccine to M cells. Recent studies demonstrated that M cells utilize several receptors to recognize and transport specific luminal antigens. Vaccination through the mucosal immune system can induce effective systemic immune responses simultaneously with mucosal immunity. How this process is regulated is largely unknown. This review aims to show a new understanding of the factors that influence the development and function of M cells; to show the molecules expressed on M cells which appear to be used as immunosurveillance receptors to sample pathogenic microorganisms in the gut; to note how certain pathogens appear to exploit M cells to inject the host; and, finally, how this knowledge is used to specifically "target" antigens to M cells to attempt to improve the efficacy of mucosal vaccines. Recently, substantial progress has been made in our understanding of the factors that influence the development and function of M cells.

The review is devoted to the analysis of pertussis incidence of children in the age group of 5–7, as well as strategies of DTP immunization with the help of the drugs in foreign countries. Mass vaccination against pertussis began in the middle of the 20th century, which contributed to a reduction in incidence and mortality rate from this infection. However, in the last decade, there has been an opposite tendency of increasing incidence of patients among children under school age, school age and adults. Atypical forms of the disease and complications due to ARVI, respiratory mycoplasmosis and cytomegalovirus infections are described in the review. Various strategies for the use of whole-cell and acellular pertussis vaccines as part of DTP drugs are described, as well as the epidemiological effect of introducing an additional booster dose of vaccine to children under school age. The expediency of revaccination of children aged 6–7 in Russia is argued, which can help to reduce the overall incidence of pertussis. The research materials related to the study of the properties of acellular anti-pertussis vaccine, such as immunogenicity and safety in comparison with whole-cell vaccine, are analyzed. The main drugs and their composition, which are used to vaccinate children against pertussis, are described in the review. It is assumed, that the increase in the incidence among children and teenagers, with the appearance of atypical forms of pertussis, is associated with a number of factors, such as the spread of new genotypes of Bordetella pertussis bacterium, emerged from mutations, as well as short duration of immunity after vaccination with acellular drugs, in comparison with whole-cell, and the use of more modern methods of detecting the pathogen. The mechanisms of the immune response due to different types of pertussis vaccines are also reviewed. It is concluded, that revaccination of children aged 6–7 with an additional fifth dose of an acellular vaccine against pertussis, as part of the DTaP instead of the Td drug, which is regulated in the National Calendar of preventive vaccinations, will have a favorable effect on the epidemic situation with pertussis infection in Russia.

 To date, clinical data have convincingly shown that C. trachomatis and C. pneumoniae infectious can cause serious diseases with severe complications and consequences. There are assumptions that the developed chronic chlamydial infection can become an important factor in the pathogenesis of the gastrointestinal tract diseases, which are manifested in the so-called post-infectious period. It is commonly known that chlamydial infection has a tropism to the cylindrical epithelium, which covers the human mucous membrane of the urethra, cervix, rectum, conjunctiva of the eyes and the throat. However, the role of the causative agents of chlamydial infections, such as C. trachomatis and C. pneumoniae, in the occurrence of the gastrointestinal tract diseases has not been studied. In order to study the possible relationship between the gastrointestinal diseases and the presence of chlamydial infection markers, we have selected a group of patients with the gastrointestinal diseases and detected antibodies to C. trachomatis and C. pneumoniae and DNA of these pathogens in blood serum, liver biopsy and bile ducts. As a result, C. trachomatis DNA in blood serum was detected in 50% of cases, and in liver biopsies — in 59.3%. A new approach has been developed in the serological diagnosis of chlamydial infection caused by C. trachomatis, which allowed for revealing diagnostic antibody titers in 51.9% of cases in this group of patients, and in the comparison group — in 11.6% of cases. Among 50% of patients, in whom DNA was revealed in blood serum, it was also revealed in 64.3% of cases in biopsy samples of gastrointestinal organs. Upon detection of C. trachomatis DNA in blood serum, antibodies to the “cultural” antigen were detected in 60.1% of cases, and with the simultaneous detection of C. trachomatis DNA in blood serum and gastrointestinal organs, they were found in 72.2% of cases. Simultaneous detection of C. trachomatis, both in blood serum and in the gastrointestinal tract, may indicate the ability of C. trachomatis to spread hematogenously and infect organs away from the primary focus of infection. The obtained data absolutely require further study in light of the identification of the relationship between the detection of the pathogen and the development of the gastrointestinal pathology. But in general, the results are not yet studied evidence of the possible gastrointestinal organs infection by C. trachomatis.

Studying of immune interlayer value to tick-borne encephalitis virus in human population of the Transbaikalia Territory resulted from natural immunization of the healthy population in 31 Municipal areas in 2011–2016 is presented. Human selections were formed proportionally to the population size in the concrete human settlement among persons of the various age and professional groups living at this territory not less than 10 years and unvaccinated against tick-borne encephalitis virus. Total 4367 blood sera were investigated. Laboratory testing for antibodies of G class to tick-borne encephalitis virus in blood sera of the human population was performed by immune-enzyme analysis using a set of reagents “VektoVKE-IgG” of Joint-Stock Company “Vektor-Best” (Novosibirsk city). The monitoring data indicated the presence of natural immunity to tick-borne encephalitis virus in the human population. Mean annual level of immune interlayer in Municipal areas varied from 3.1 to 52.7% (in Transbaikalian Territory — 13.1±0.51%). High level (from 20 to 52.7%) was characteristic for mountain-taiga-forest-steppe zone (Krasnochikoisky — 23.8±3.36%; Uletovsky — 52.4±4.48%; Gazimuro-Zavodsky — 29.4±4.94% districts) and mountain-tundra-taiga zone (Tungokochensky district — 20.0±3.58%). In steppe zone the level of immune interlayer was lower and observed in districts with elements of south-taiga larch and pine forests. It was established that levels of the immune interlayer in human population living in mountain-taiga-foreststeppe and mountain-tundra-taiga zones authentically higher than in steppe area — t = 3.8; Р < 0.001 and t = 2.27; Р < 0.05, respectively. Distinctions in the value of the immune interlayer between mountain-tundra-taiga and mountain-taigaforest-steppe zones were non-authentic: t = 0.1; P > 0.05. Active circulation of tick-borne encephalitis virus was accompanied by authentic (Р < 0.01) increase of the interlayer in persons with the virus antibodies in 2014–2016 (15.8±0.69%) in comparison with 2011–2013 (9.7±0.78%). Also it was noted in all landscape zones: in steppe zone the increase was to 42.8% (P > 0.05); in mountain-taiga-forest-steppe — to 61.3% (P > 0.05); in mountain-tundra-taiga — to 150.0% (Р < 0.01). It was not possible to reveal correlation between the recourse for medical aid and the value of immune interlayer in the population formed as a result of latent immunization. Results of the population immunity studying essentially expand our knowledge about the condition of the natural foci and dynamics of development of the epidemic processes in it, and can be used at planning of the preventive actions.

It is known that resistant Enterobacteriacaestrains can be a part of human microbiota. While colonizing the body of a healthy person, resistant strains do not cause diseases, while the healthy person becomes carrier and source of such strains or the resistance genes to the external environment. The aim of the study was to compare the antimicrobial susceptibility of opportunistic Enterobacteriacaeisolated from feces of residents of the Republic of Guinea (121 strains) and Russia, Saint Petersburg (897 strains). The antimicrobial susceptibility was determined by the disc-diffusion method according to the national russian Clinical Guideline “Antimicrobial susceptibility testing of microorganisms” using discs and Muller-Hinton agar manufactured by Oxoid. Resistance mechanisms were tested in bacterial strains non-susceptible to beta- lactams using confirmatory phenotypic and molecular tests. Resistant strains were detected significantly more often in the intestinal microbiota of residents of the Republic of Guinea, than of Saint Petersburg (83.5 and 28.7 per 100 strains studied, respectively), including strains with multidrug resistance (47.9 and 11.1, respectively). The residents of the Republic of Guinea have a high frequency of isolation of Enterobacteriacaeresistant to “old” antibiotics, which were often used in the 1970s, but rarely used in Russia and other European countries: tetracyclines (63.2), trimethoprim/ sulfamethoxazole (59.5) and aminopenicillins (48.4). The frequency of detection of strains resistant to modern clinically significant antibiotics (extended spectrum cephalosporins, f luoroquinolones, aminoglycosides) was the same in residents of the Republic of Guinea and Saint Petersburg. Resistance to beta-lactams in Enterobacteriacaestrains isolated both from the residents of the Republic of Guinea and from Saint Petersburg is due to the same mechanism — the production of beta-lactamases — broadspectrum and extended spectrum- of various genetic families. This data corresponds to world global trends in the antimicrobial resistance in Enterobacteriacae: resistance to aminopenicillins in E. coliis due to the production of broad-spectrum beta-lactamases TEM-1 (80% of strains from residents of the Republic of Guinea and 86.5% from Saint Petersburg), and resistance to extended spectrum cephalosporins — the production of extended spectrum beta-lactamases of the CTX-M1 genetic group (72.7 and 67.6% respectively). Our research has shown that the intestinal microbiota of the inhabitants of different continents (Europe and Africa) contains the strains (Klebsiella pneumoniae and Escherichia coli) resistant to clinically significant antibiotics (beta-lactams) due to single mechanism globally spread in Enterobacteriacae.

Purpose: efficiency isothermal amplification (LAMP) at inspection of patients with whooping cough in clinical conditions. Materials and methods. Examination of 262 patients aged from 0 months up to 30 years hospitalized in Infectious diseases clinical hospital No. 1 of the Moscow Department of Healthcare is conducted. Clinical specimens (pharyngeal swabs) were collected according to MR 3.1.2.0072-13. Extraction DNA of B. pertussiswas carried out by means of the АmplyPrime® DNA-sorb-AM. Identification of specific fragments of a genome of B. pertussiswas performed by PCR-real time by means of the АmplySens ® Bordetella Multi-FL set (a comparison method) and by the LAMP with a phoresis and intercalating dye. Results.When using of the optimized method LAMP the DNA of B. pertussisis found at 252 (96.2%) patients. The method was effective at any forms of whooping cough —DNA of B. pertussisis found in all patients with a severe form, in 95.8% of cases — in patients with medium-weight and in 95.3% of cases — in patients with an easy form. The DNA of B. pertussisis found in clinical specimens received from patients on different terms from the beginning of a disease — from 92.3% on the 1st week up to 96% of cases — on the 5 th and more weeks of a disease. The DNA of B. pertussisis found in high percent of cases (96.7–95.9%) and did not depend on acceptance of antibacterial therapy. Children till 1 year are the main age group which is subject to hospitalization at suspicion of whooping cough and in which the highest risk of development of complications and severe forms of a clinical disease. At inspection of 169 children from 0 to 12 months by means of the optimized method LAMP, DNA of B. pertussisit is found in 98.6% of cases in children of patients with whooping cough aged from 0–3 months, in 98.4% of cases — children of 4–7 months and in 94.6% of cases — at children have 8–12 months. The efficiency of detection of DNA of B. pertussisat patients with whooping cough of children aged till 1 year by means of the optimized LAMP method was 97.6%.

The aim of the study was to examine the features of phenotype and the levels of NAD(P)-dependent dehydrogenases activity of blood neutrophils in the prognosis of abdominal sepsis development in patients with widespread purulent peritonitis (WPP). 50 patients with WPP of community and hospital origin in the pre-operative period were examined. From the 5th to the 10th day of the postoperative period 35 patients (70%) had developed abdominal sepsis, 15 patients (30%) had absence of complications. As controls 67 respect healthy people were examined. The research blood neutrophils phenotype was performed by f low cytometry using a direct immunofluorescence whole peripheral blood. The levels of surface receptor expression was assessed by the mean fluorescence intensity. The NAD(P)-dependent dehydrogenases activity in the blood neutrophils studied using bioluminescence method. It was established that the inflammatory reaction in patients with WPP is characterized by neutrophilia and changes in the phenotype of blood neutrophils. The markers of the subsequent development of sepsis in WPP are less pronounced (in comparison with the indices of uncomplicated patients), an increase in the number of neutrophils, a decrease in the HLA-DR + -cell count against the background of a high level of neutrophils expressing a high affinity receptor for IgG. The patients without subsequent complications had the number of neutrophils with CD23 receptor expression is increased. Metabolism of neutrophils in patients with WPP is characterized by a decrease in the intensity of plastic processes due to low activity of glucose-6-phosphate dehydrogenase and an imbalance in the activity of the enzymes of the mitochondrial compartment. A feature of the neutrophil metabolism in patients with WPP without subsequent development of sepsis is a high activity of anaerobic lactate dehydrogenase reaction and a decrease in the activity of NADP-dependent decarboxylating malate dehydrogenase. The patients with WPP with subsequent development of sepsis had a high level of NAD-dependent out flow of substrates from the tricarboxylic acid cycle on the amino acid exchange reaction via glutamate dehydrogenase which can affect the activity of aerobic respiration of blood neutrophils. The established differences in the phenotype and activity of enzymes in the blood neutrophils in patients with WPP depending on the subsequent development of sepsis determine the possibility of creating a method for predicting the development of complications and developing immunoactive therapy in the postoperative period of WPP.