REVIEWS
Sarcoidosis is a systemic inflammatory disease of unknown etiology, characterized by the formation of epithelioid cell granulomas, multisystem lesions with a certain frequency of involvement of various organs, mainly the lungs (up to 90% of cases). Over the past decade, significant progress has been made in understanding the pathogenesis of sarcoidosis, the important role of immunological, genetic and environmental factors in the development of this pathology has been established. It is believed that the leading mechanism in the pathogenesis of sarcoidosis is the aberrant activation of the innate and adaptive immune response to unidentified antigen(s), which leads to the development of granulomatous inflammation and the formation of granulomas. However, despite the huge number of studies that has been carried out, the mechanisms and signaling pathways that control the development of the inflammatory process during the formation of granulomas and the progression of pathology have not been fully determined.
This literature review examines the important role of various cytokines and T helper subpopulations in sarcoidosis. Particular attention is paid to the cells of innate immunity – macrophages in the pathogenesis of this disease. These cells play a key role in the formation of sarcoid granulomas and in the pathogenesis of sarcoidosis. The macrophage population is characterized by plasticity and functional heterogeneity. In response to various signals from the microenvironment, macrophages are able to acquire certain phenotypes. The review considers the issues of polarization of macrophages, changes in the phenotype of these cells to subpopulations M1 (M1 phenotype; classically activated; pro-inflammatory) and M2 (M2 phenotype; alternatively activated, anti-inflammatory). These two cell populations are characterized by the expression of different markers on their surface, which allow these cells to differentiate from each other. The analysis of literature data on the levels of key polarizing cytokines for macrophages and cells-producers of these cytokines that patients with sarcoidosis have, in acute and chronic course of the disease, was carried out.
Important aspects of the alternative activation of macrophages of the M2 phenotype and their division into subtypes: M2a, M2b, M2c, M2d are noted. The features of various subtypes’ activation of macrophages in this granulomatosis and their importance in the development and progression of pathology are considered. Studying the role of macrophages’ phenotypes, understanding the mechanisms by which the phenotypes of these cells are activated and modulated in various microenvironmental conditions, can contribute to the development and implementation into clinical practice of new therapeutic approaches for the treatment of sarcoidosis and many other forms of pathologies.
The mother and fetus incompatibility due to Rh-factor, blood group or other blood factors can lead to hemolytic disease of the fetus and newborn (HDN). HDN is a clinical disease condition of the fetus and newborn as a result of hemolysis, when maternal IgG alloantibodies cross the placenta and destroy the red blood cells of the fetus and newborn. The child disease begins in utero and can dramatically increase immediately after birth. As a result, hyperbilirubinemia and anemia develop, that can lead to abortions, serious complications, or death of the neonates in the absence of proper therapy. The range of HDN has changed significantly now compared to previous decades. Half a century ago, HDN was considered an almost complete synonym of RhD-alloimmunization, and this was a frequent problem for newborns. By now due to the high effective of Rh-conflict prevention, immunological AB0-conflicts have become the most common cause of HDN. The review aimes to one of the main causes of jaundice and anemia in neonates at present, i.e. HDN due to immunological AB0-conflict of mother and newborn (AB0-HDN). The main participants of the AВ0- incompatibility mother and child are considered, namely A- and B-glycans, as well as the corresponding anti-glycan alloantibodies. Close attention is paid to the structure features of glycan alloantigens on the red blood cells of the fetus and adult. The possible correlation of the frequency and severity of HDN with the blood group of mother and child, as well as with the titer of maternal alloantibodies, has been considered. The influence of immunoglobulin G subclasses on the AB0-HDN development has been evaluated. In most cases, AB0-HDN appear when the mother has the blood group 0, and the fetus has the group A (subgroup A1) or the group B. Other rare incidences of AB0-incompatibility with severe course are occurred. As a whole the etiology of AB0-HDN is complex and the HDN severity is influenced by many factors. The authors have analyzed statistical data, as well as the prevalence of AB0-incompatibility and AB0-HDN in various regions of the world. Current approaches to the diagnosis of AB0-HDN are discussed in addition. By now the problems of AB0- HDN occurrence and developing of ways to overcome this disease remain relevant.
Lassa virus (LASV) is classified into genus Mammarenavirus of Arenaviridae family. This virus is etiological agent of Lassa fever (LF) which is widespread in Africa. On average, in four out of five infected people, LF occurs without symptoms. The annual incidence ranges from 100,000 to 500,000 registered clinical cases, at a mortality rate of 1-2%. Among hospitalized patients with severe symptoms of hemorrhagic fever, this figure may be from 14 to 89.5%. Signs of an adverse outcome in LF are open bleeding and disorders of CNS (convulsions, tremor, disorientation and coma). Death occurs from multiple organ failure. Severely ill people recover slowly and may have relapses and complications such as pneumonia, myocarditis, psychosis, and hearing loss.
Transmission of the virus in endemic territories occurs by alimentary way, air-dust and airborne droplets from a zoonotic source – rodents of the species African multimammate rat (Mastomys natalensis), by accidental contacts of people with their secretions (urine, feces, saliva) as well as when butchering carcasses and eating rodents. These animals are characterized by asymptomatic carrier and life-long persistence of the virus. Cases of transmission of the virus from person to person through the blood or other body fluids of patients are described. A sick person is contagious for two months, because the virus circulates in the blood despite high levels of antibodies. Infection of medical staff occurs during emergency surgical operations, or when the rules of contact precautions are not observed. Currently, with the ongoing LF outbreak in Nigeria, since 2016, hospitals have registered mortality rates of 22 and 8% for patients and health workers, respectively. During 1969-2016, 33 imported cases of this disease were described from West Africa to non-endemic territories (in the USA, Canada, Great Britain, the Netherlands, Germany, Israel and Japan). The mortality rate among these patients was 39%.
The lack of prophylactic vaccines and specific therapeutic drugs is the major challenge for the prevention of LF. Thus, this review considers biological models (cell cultures and animals) that are suitable for studying the pathogenesis of this disease, preclinical studies of the specific activity and harmlessness of candidate vaccines, as well as options for these developments based on the platforms such as inactivated LASV and its DNA, the reassortant of Mopeia arenavirus, and measles virus attenuated strains, recombinant and replication-defective viruses (smallpox vaccine, Venezuelan equine encephalitis, bovine vesicular stomatitis, adenovirus of chimpanzee) and virus-like particles.
ORIGINAL ARTICLES
Numerous studies over last decade have shown that functional capacity of neutrophil granulocytes (NG) determines the course and outcome of many diseases. Identification of phenotypic variants of functionally significant NG subpopulations is a new approach allows us to assess the adequacy or deficiency of NG involvement into infectious inflammation processes at molecular level. An opportunity of reorienting a deficient NG subpopulational phenotype in purulent inflammatory diseases due to the rearrangement of the receptor set induced by various immunotropic substances may serve as a key to recovery of normal NG functioning.
Our aim was to study the effect of glucosaminylmuramyldipeptide (GMDP) under in vitro conditions upon the phenotypic profile of four functionally significant subpopulations, i.e., CD62L+CD63-NG, CD62L+CD63+NG and CD64-CD32+CD16+CD11b+NG, CD64+CD32+CD16+CD11b+NG, along with assessment of expression density of appropriate membrane molecules and NG microbicidal activity in the children with purulent inflammatory diseases. 90 samples of peripheral blood (PC) were taken from children 2 to 4 years old, including 12 children with minor purulent infection (MPI), and 7 children were studied as conditionally healthy controls. Their peripheral blood was incubated for 60 minutes at 37 °C with GMP (10-6 g/l). Using flow cytometry technique, the relative numbers of some NG subpopulations, i.e., CD64-CD16+CD32+CD11b+NG, CD64+CD16+CD32+CD11b+NG, CD62L+CD63-NG, CD62L+CD63+NG were evaluated, and the phenotype features of each subpopulation were investigated according to the density of appropriate membrane molecule expression (MFI). In parallel, phagocytic and microbicidal activity of NG was tested in these study groups. The obtained data indicate for presence of for distinct NG subpopulations, both in healthy children and in children with MPI. We have revealed phenotypic transformation of the four studied NG subpopulations from MPI patients including disturbed phagocytic and microbicidal functions of the cells. Using of this in vitro system, we have shown that the transformed phenotype of the four functionally significant NG subpopulations of MPI patients was re-arranged under GMDP treatment. At the same time, the number of CD62L+CD63+NG and CD64-CD32+CD16+CD11b+NG subpopulations was increased, along with decreased amounts of CD64+CD32+CD16+CD11b+NG and CD62L+CD63-NG subpopulations, being accompanied by restoration of microbicidal activity of NGs.
The obtained data allow us to accomplish current understanding of immunotropic effects of GMDP, and to extend the potential scope of its experimental and clinical application. The new data on GMDP effects revealed by in vitro system, i.e. phenotype rearrangement of functionally significant NG subpopulations CD64-CD16+CD32+CD11b+, CD64+CD16+CD32+CD11b+, CD62L+CD63-, CD62L+CD63+ in atypical purulent inflammatory diseases in children, may be used in the future in order to develop innovative strategies of immunotherapy aiming for correction of NG dysfunction in children with MPI.
Our aim was to evaluate diagnostic and pathogenetic significance of plasma cytokines (IL-20, IL-23, IL-10, TNFα, IFNγ) in the patients with various clinical and histological variants of sclerotic lichen and to assess opportunity for their use as effectiveness criteria of immunotherapy for this disease using a drug based on eukaryotic deoxyribonucleic acid (Derinat). The prospective cohort study included assessment of the clinical manifestations (itching and dyspareunia) and measurement of blood cytokine contents (IL-20, IL-23, IL-10, TNFα, IFNγ) in women (n = 114) with various clinical variants of sclerotic lichen (atrophic, sclerotic and sclerotic-atrophic) before and after immunotherapy with a nucleic acid-based drug (Derinat). Derinat was chosen due to the fact of being an agonist of Toll-like receptors, and a number of immunoregulatory effects, including the ability to modulate cytokine production and to exert a positive influence upon regeneration processes. In addition, based on visual inspection, vulvoscopy and morphohistochemical examination results (evaluation criteria: skin thickness, number of collagen fibers, severity of fibrosis and sclerosis, etc.), the corresponding subgroups were classified within the II group, i.e., 2.1 (minimal sclerotic signs, n = 14), and 2.2 (pronounced sclerotic signs, n = 20). The control group consisted of conditionally healthy women, without history or presence of vulvar pathology (n = 30), with an age ranging from 20 to 50 years. Along with cytokine assessment by enzyme immunoassay, the study used the data of clinical examination (anamnesis collection, examination, palpation, vulvoscopy), as well as complex morphohistochemical evaluation of vulvar tissues. In atrophic variant, we have observed an increase in plasma IL-23 content, along with decreased TNFα; in lichen sclerosis, a maximal increase in IL-20, IL-23, and IFNγ was revealed; in sclerotic form of sclerotic lichen variant with severe sclerotic features, maximally enhanced IL-20, IL-23, TNFα, IFNγ, along with minimal levels of IL-10 was registered, as compared with other groups. Immunotherapy using Derinate resulted into significant reduction in the clinical manifestations in sclerotic lichen, i.e., itching of the vulva and dyspareunia, as well as normalization of cytokine indexes. Our studies have demonstrated an opportunity of using plasma concentrations of IL-20, IL-23, IL-10, TNFα, IFNγ as biomarkers of sclerotic lichen variants, and as laboratory criteria for efficiency of immunotherapy.
Sarcoidosis is a polysystemic inflammatory disease of unknown etiology, morphologically related to the group of granulomatosis, with heterogeneous clinical manifestations and outcomes. Immune cells, in particular T helper cells, are attracted to lung tissue and/or other organs by chemokine gradients and play an important role in the granuloma formation. T helper cells migrate from peripheral blood to the tissues due to expression of CXCR3 chemokine receptor on their surface. It interacts, e.g., with CXCL9/MIG, CXCL10/IP- 10, and CXCL11/I-TAC. Our study was aimed for determining the levels of CXCL9/MIG, CXCL10/IP-10, CXCL11/I-TAC chemokines in peripheral blood of the patients with sarcoidosis, depending on the features of their clinical course before administration of immunosuppressive therapy. We studied peripheral blood plasma samples of the patients with sarcoidosis (n = 52). In 37% (19/52), they exhibited acute clinical manifestations, and 63% (33/52) had chronic sarcoidosis. The control group included peripheral blood samples from healthy volunteers (n = 22). The chemokine concentrations (pg/ml) were determined by multiplex analysis using xMAP technology (Luminex), and Milliplex MAP test system (Millipore, USA). In the patients with sarcoidosis, significantly higher levels of chemokines were shown relative to healthy volunteers: CXCL9, 4013.00 pg/ml vs 1142.00 pg/ml (p < 0.001); CXCL10, 565.90 pg/ml vs 196.60 pg/ml (p < 0.001); CXCL11, 230.20 pg/ml vs 121.10 pg/ml (p = 0.018). Plasma concentrations of CXCL9 and CXCL10 were significantly increased both in blood samples from patients with acute and chronic sarcoidosis compared to healthy volunteers, p < 0.001. The level of CXCL11 chemokine was significantly increased only in the patients with chronic sarcoidosis, compared to the healthy volunteers: respectively, 251.50 pg/ml and 121.10 pg/ml (p = 0.044). The levels of this chemokine correlated with the activity of angiotensin-converting enzyme (ACE), with r = 0.374; p = 0.042. The ACE level in sarcoidosis is considered a clinical and laboratory index of the disease activity. In acute sarcoidosis, the level of CXCL11 chemokine was not significantly higher than in healthy individuals, whereas the CXCL9 chemokine content was significantly increased and correlated with ACE activity (r = 0.762; p = 0.037). The level of CXCL9 chemokine was significantly decreased in patients with signs of fibrosis as compared with fibrosis-free patients (1839.88 pg/ml vs 4375.52 pg/ml, p = 0.035). Significantly higher levels of CXCL9 were detected in cases of systemic sarcoidosis, i.e. 6036.84 pg/ml, as compared with 1927.44 pg/ml in the patients without these signs (p = 0.018). Evaluation of clinical and laboratory diagnostic characteristics for plasma chemokine levels in sarcoidosis patients allowed to assess their sensitivity and specificity. The respective values were as follows: in acute sarcoidosis: for CXCL9, 84% and 95%; for CXCL10, 84% and 95%; for CXCL11, 74% and 59%. In chronic sarcoidosis, the respective values for CXCL9 were 82% and 72%; for CXCL10, 91% and 77%; for CXCL11, 79% and 55%, respectively. Thus, the determination of plasma CXCL9, CXCL10, and CXCL11 chemokines in sarcoidosis allows of understanding their role in development of the disease, e.g., recruitment of T helper cells from peripheral blood to the lung tissue, and granuloma formation. Clinical and immunological comparisons of CXCL9 levels in the peripheral blood of patients and characteristics of the clinical course of sarcoidosis indicate to the role of this diagnostic parameter for assessing the disease activity, signs of lung fibrosis, and systemic manifestations in this disease.
Background: Hepatitis B is the main infection of the injured liver for humans. Inflammation of the liver is caused by hepatitis viruses may lead to cirrhosis and hepatocellular carcinoma. The B-cell stimulatory factor 2 (BSF-2) is one of the cytokines that affect the regulation and differentiation of the human immune response. Objective: this report aims to estimate the BSF-2, GPT, and GOT levels in patients’ serum with different stages of hepatitis B compared with healthy control. Methods: This study assessed 52 patients presumably with acute and chronic cases who have HBsAg positive. BSF-2 was detected using ELISA assay. Biochemical parameters were determined using kits of an automated analyzer. SPSS version-16 software was used for statistical analysis. Results: Acute hepatitis B patients had shown elevation in BSF-2 level more than of chronic hepatitis B. GPT and GOT levels elevated in the acute hepatitis group more than of the chronic hepatitis group. We reported a significant value between BSF-2, GOT, and GPT levels. We didn’t score an association between patient’s age and cases groups of hepatitis. Conclusion: our data confirmed increasing of BSF-2 levels with the increase of GOT level more than GPT level with acute hepatitis B. BSF-2, GPT and GOT levels are varied in different courses of acute and chronic HBV. We surmised that the elevation of BSF-2 levels designates liver injury of patients with acute HBV.
Glaucoma is one of the leading causes of irreversible blindness worldwide, being an age-related disease. Its pathogenesis still is not fully understood. A particular interest is attracted to evaluation of the cytokine concentrations in the trabecular meshwork cell culture, and in the aqueous humor (AH) taken from the same patient, since such data may allow to describe more completely the glaucomatous trabecular changes and to clarify the mechanisms of intercellular interactions in pseudoexfoliative (PEX) glaucoma. The purpose of this study was a comparative analysis of cytokine contents in AH and in trabecular tissue (TT) supernatants in the patients with PEX glaucoma. The study included 23 eyes of patients with PEX glaucoma. The material studied was AH and supernatant of TT cell culture. The cytokine concentration was measured using a flow cytofluorimeter FacsCantoII (BD, USA) using the CBA method. SPSS version 19 software (IBM, USA) was used for the statistical data processing. Concentrations of cytokines (TNFα, IFNγ, IL-1β, IL-6, IL-8, IL- 10, VEGF, GM-CSF) were determined in AH and in the TT supernatant for each of the patients with PEX glaucoma. Only IL-6 and VEGF concentrations in AH were higher than those in the TT supernatant in patients with PEX glaucoma. The IL-6 concentration positively correlated with the VEGF and IL-8 concentrations in the TT supernatant. Correlations between other cytokines in the TT supernatant and AH were also identified and analyzed. Multiple regression analysis revealed that the duration of glaucoma and the IFNγ and TNFα concentrations in AH may have a significant influence on the corneal endothelial cells, being associated with density reduction in patients with PEX glaucoma. The correlation analysis did not reveal any links between other clinical data (corneal thickness in the optical center, IOP level, age) and the cytokine concentrations in the studied tissues. The obtained results suggest that only simultaneous analysis of the cytokine concentrations in the TT supernatant and AH taken from the same patient may provide a more complete description of the cytokine imbalance and pathological processes occurring in the trabecular meshwork in PEX glaucoma patients. It has been shown that the changing cytokine ratios observed in PEX glaucoma may be associated with development of uniform structural and functional changes in all tissues of the anterior eye segment.
Allergy is a sufficient social and economic issue of modern times. Altered immunity in allergic disorders is based, mainly, on the lymphocyte disturbances.
Immune characteristics depend both on populations and subpopulational profile of immune cells, and on intrinsic intensity and specific features of theirintracellular metabolism. Interest to studies of intracellular metabolism of lymphocytes id determined by high-scale energetic and plastic processes aimed for support of immune homeostasis. The aim of this study was to evaluate the state of intracellular metabolism in peripheral blood lymphocytes from the patients with respiratory allergies of different genesis and respiratory affection.
The study included patients with various clinical variants of respiratory allergy (n = 152) at the age of 21 to 63 years old, and virtually healthy blood donors (n = 209), comparable for age and sex. Within these cohorts, we have separately analyzed, e.g., respiratory atopy (atopic rhinosinusitis and atopic bronchial asthma), as well as respiratory pseudoatopy (polypous rhinosinusitis and asthmatic triad). Allergic disorders of upper respiratory ways were diagnosed in a complex clinical examination by allergologist/immunologist and otorhinolaryngologists. Bronchial asthma verification was based on current GINA criteria (2014). We used m standard common clinical methods and specific allergological diagnostics, e.g., allergological anamnesis, skin prick tests, with different non-infectious allergens, measurement of total and specific IgE’s with ELISA method. The parameters of intracellular metabolism of peripheral blood lymphocytes were determined with bioluminescent technique with bacterial luciferase. Actifity of NAD(P) and NAD(P)H enzymes was measured.Dehydrogenase activities in lymphocytes were expressed as enzyme un its (EU, 1 unit = 1 mcmol/min) per 104 cells.
Certain changes of intracellular activities in peripheral lymphocytes are revealed, dependent on genesis and origin of allergic inflammation, and affection level of respiratory ways. In respiratory atopy (atopic rhinosinusitis and atopic bronchial asthma), irrelevant on the level of respiratory affection, the activities of intracellular enzymes suggest increased plastic processes that are maximally pronounced in atopic bronchial asthma. In respiratory pseudoatopy (polypous rhinosinusitis and asthmatic triad) the metabolic changes of lymphocytes presume activation of both plastic and energetic processes, with decreased intensity in asthmatic triad condition. Independent on genesis of respiratory allergic inflammation, we have determined low activity of NAD(P)-dependent glutamate dehydrogenase, and NAD(H)-dependent lactate dehydrogenase in allergic inflammation of upper respiratory ways (atopic rhinosinusitis and polypous). Its activity is statistically higher in bronchial asthma (atopic bronchial asthma and asthmatic triad.
The first studies were published on the possible pathogenetic role of so-called ectopically localized taste receptors in bronchial asthma. The receptors for bitter and sweet taste, may, apparently, have opposite functions, but in available literature there is no data on the balance of sensitivity for bitter and sweet tastes in the same patients with bronchial asthma. The aim of the present work is to simultaneously assess the sensitivity of canonical lingual receptors to bitter and sweet taste in the same patients with different clinical variants of bronchial asthma by methods applicable in wide clinical practice. 16 healthy persons and 35 patients with bronchial asthma were examined at the M.V. Chernorutsky Clinics of Hospital Therapy at First St. Petersburg State I. Pavlov Medical University. The sensitivity for bitter taste was assessed using The Frey Scientific 569885 PTC Taste Paper test strip kit containing phenylthiourea solution. Sucrose solutions at concentrations of 0.3; 0,4; 0,5; 0,6; 0,7; 0,8; 0,9 % for determination of individual value of taste thresholds to sweet taste were used. The bitter-to-sweet taste sensitivity balance was assessed on the basis of an original “bitter/sweet taste sensitivity” index. The highest values of index of bitter/sweet taste was found in the allergic variant of bronchial asthma: its values are significantly different from those in healthy persons only at low sucrose concentrations (0.3-0.4%). The factor analysis revealed an association between taste imbalance (a shift towards high sensitivity to sweet taste) and key characteristics of bronchial asthma, including severity of bronchial asthma course, duration of inhaled glucocorticosteroid use and inefficiency of β2-agonists use at pre-clinical stage. It has been revealed by gustometry that in the allergic variant of bronchial asthma there is a decreased sensitivity for bitter test substance (phenylthiourea), along with higher sensitivity for sweet taste (sucrose).
At the present time, the efforts of many research groups around the world are aimed at finding new factors triggering the allergic sensitization process linked with IgE synthesis to harmless allergens. According to the recent data, production of tissue cytokines is induced in tissue cells by alarmins, thus, in turn, eliciting pro-allergic immune response. Previously we have shown that β-alanine could be a potential alarmin capable to stimulate production of tissue cytokines. The aim of this work was to determine the impact of β-alanine on humoral immune response in low-dose allergy model. BALB/c mice were immunized by recombinant Asp f 2 protein or commercial ovalbumin (OVA) in the withers 3 times a week with or without β-alanine supplementation. To determine the mechanism of β-alanine effect, α-L-alanine, an isomer which is not MrgD receptor ligand, and β-aminoisobutyrate with β-alanine-like affinity to MrgD ligand, were compared. According to our data, β-alanine stimulated specific IgE and IgG1 production in a short-term course (7 immunizations) and enhanced antibody affinity after long-term (14 immunizations) protocol in the case of low-immunogenic protein Asp f 2. In the case of high-immunogenic OVA protein, the impact of β-alanine was significant only upon antibody affinity. Hence, β-alanine accelerates specific IgE production in the case of low-immunogenic protein. The impact of β-alanine on specific IgE production was not linked to specific MrgD receptor activation, because β-aminoisobutyrate, which is the other ligand of this receptor, did not have a similar effect upon humoral immune response. The effect of β-alanine on IgG1 production seems also independent of MrgD receptor, since the common proteinogenic amino acid α-L-alanine also enhanced specific IgG1 production. The effect of β-alanine on humoral immune response could be linked to its non-specific action, e.g., due to its ability to induce oxidative stress through blocking taurine transporter, or due to its ability to stimulate cellular metabolism.
SHORT COMMUNICATIONS
Permafrost is a unique ecosystem characterized by consistently negative temperatures. It has been shown that microorganisms can be there in a state of hypometabolism or anabiosis during geological time. It is known that microorganisms occupy a wide habitat due to the presence of multifunctional systems of adaptation and communication. One of the manifestations of these systems is the production of secondary metabolites (MBs), which include signaling molecules that do not have strict species specificity. The biological activity of signaling molecules largely depends on the number of bacterial cells and the temperature of their cultivation.
In this work we used secondary MBs of Bacillus sp. from Permafrost obtained at different temperatures of microorganism cultivation (at -5 °C – “cold” MBs and at 37 °C – “warm” MBs) in doses of 0,05 × 106 (small dose) of microbial cells (m.cl.) in ml of saline or 500 × 106 (high dose) m.cl./ml. The influence of MB of Bacillus sp. for the TNFα, IL-1β, IL-8, IL-2, IFNγ, IL-4 and IL-10 production by human peripheral blood mononuclear cells (MNC) in supernatants of 24-hour cell cultures was estimated by ELISA whith using the “VectorBEST” test system (Russia) on a LUCY-2 (ANTHOS) spectrophotometer (Austria).
It was found that the activity of synthesis by human MNC of the main spectrum of cytokines significantly increased (p < 0.01 for all indicators) under the influence of MB Bacillus sp. regardless of the temperature of their cultivation and the dose of bacteria. The exception was IL-8, the level of which under the influence of a high dose of “warm” MBs didn’t differ from the control. Compared to PHA the cytokines synthesis by MNC depended on the dose and the temperature of obtaining of MBs. Thus, under the influence of “warm” MBs the level of TNFα was significantly lower than its level under the influence of PHA regardless of the dose. Regardless of the temperature of obtaining metabolites the level of IL-8 under the influence of metabolites from a dose of 500 × 106 m.cl. was reduced relative to the PHA group. Comparison of the influence of “warm” and “cold” MBs of Bacillus sp. showed that small doses of “cold” metabolites to a greater extent stimulate the synthesis of pro-inflammatory cytokines (TNFα, IL-1β, IL-8, IFNγ). High doses of “heat” metabolites of Bacillus sp. to a greater extent they activate human MNCs for the synthesis of anti-inflammatory cytokines (IL- 4 and IL-10). Considering that TNFα, IL-1β and IL-10 are cytokines of systemic action and are responsible not only for the activation of the immune system, but also for the mobilization of other regulatory systems of the organism, it can be assumed that the secondary metabolites of microorganisms from Permafrost will be efficient as a substrate for the development of new immunomodulators and adaptogens in the future.
This study is aimed to investigate the effect of female autoserum on the HLA-DR expression in various subpopulations of lymphocytes obtained from spouses with children with sporadic congenital heart defects without chromosomal diseases. 78 married couples with children with congenital heart disease were included in the study group. The control group was formed from 35 married couples with healthy children. The immune response in a mixed culture of lymphocytes of spouses was evaluated by an increased HLA-DR expression in a mixed culture in relation to spontaneous cultures of lymphocytes. Primary staining of female and male lymphocytes by monoclonal antibodies to CD45 conjugated with various fluorescent dyes (PC-5 and PC-7) was performed to assess the immune response of female lymphocytes to male ones and vice versa. The activating effect of female autoserum on all subpopulations of female lymphocytes simultaneously occurred significantly less frequently in the study group compared to the control. The control group was characterized by the domination of the positive effect of female autoserum on HLA-DR expression for all subpopulations of female lymphocyte. For all female lymphocytes having HLA-DR molecule on its membrane, the blocking effect of female autoserum in the study group was significantly more expressed in relation to the control group. Thus, the effect of female autoserum is manifested in relation to the HLA-DR expression on its own lymphocytes, but not on the lymphocytes of the spouse.
The changing states of T cell populations responsible for the chronic course of allergic inflammation and diseases, including allergic bronchial asthma, are not yet sufficiently characterized. The aim of this study was to detect phenotypic changes in the CD45RA/CD45RO positive T lymphocytes and the level of regulatory cytokines (TNFα, IFNγ, IL-4, IL-6, IL-8, IL-10, IL-13, IL-17А, IL-17F) in allergic respiratory diseases (ARD) in children. In blood of 90 children aged 3-11 (60 children with ARD and 30 healthy peers) were studied of the immune cellular populations and cytokine indices. The levels of IL-4, IL-8, IL-10, IL-13, IL-17A and IL-17F in blood serum of children with bronchial asthma and allergic rhinitis differed from appropriate indices in control group (p = 0.001). The quantity of CD3+CD8+CD45RACD45RO+ cells, T helpers (p < 0.05) and Th effectors simultaneously expressing both isoforms of the CD45RA+ and CD45RO receptor in peripheral blood of children with ARD significantly exceeded those in control group (p < 0.001). In healthy children, Th17 population (CD3+CD4+CD196 lymphocytes) comprised 9.49±1.6% of CD3+CD4+ of cells, the number of such lymphocytes was significantly increased to 14.5±0.77 in children with allergic diseases (p < 0.001). Absolute numbers of Th17+ cells were 93.0±9.30 and 127,0±72.0 cells/µl respectively (p = 0.002). Indicators of CD4CD45RO positive memory cells in children with ARD was determined as significantly lower (p < 0.001), whereas quantity of CD3+CD19+ proved to be higher (p < 0.05) than in healthy peers. Absolute counts of these cells did not differ between the groups. The number of CD8+CD45RO+T lymphocytes was significantly higher in children with allergic diseases (p < 0.025). This research shows that the quantitative ratio of CD3+CD8+CD45RA+ and CD3+CD8+CD45RO+ populations of T cells, and increased levels of cytokines, synthesizable via Th2 and Th17, in peripheral blood may be helpful for understanding genesis of allergic respiratory diseases, and extends our knowledge on immune mechanisms of allergic disorders for individualization of therapeutic programs.
The aim of the study was to analyze n-3 and n-6 polyunsaturated fatty acid (PUFA) profile of blood leukocyte cytomembranes in mild and moderate COPD, and to establish possible role of these fatty acids in COPD progression. The study involved 110 patients with mild disease (n = 60) and moderate COPD (50 patients), at average age of 57.5±4.8 years old. The control group consisted of 32 practically healthy non-smoking people with normal pulmonary function (average age 42.0±3.4 years). The immunological study included flow cytometric determination of blood immune cell subpopulations, i.e., T lymphocytes (CD3+), T helper cells (CD4+), cytotoxic T lymphocytes (CD8+), and B cells (CD19+) using Becton Dickinson machine (USA). Fatty acid methyl esters redissolved in hexane were analyzed using “Shimadzu GC-2010” gas-liquid chromato-graphic system (Japan). Analysis of the polyunsaturated fatty acid profile of leukocyte membranes in COPD patients revealed a reduced concentration of essential linoleic acid (18:2n-6) regardless of the disease severity. The leukocyte membrane levels of the long-chain n-6 PUFAs, such as dihomo-γ-linolenic acid (20:3n-6), arachidonic acid (20:4n-6), and docosatetraenoic acid (22:4n-6), were elevated in patients with COPD compared with the control group. However, the concentration of the described above n-6 PUFAs in leukocyte membranes was increased in patients with moderate COPD compared to the patients with mild COPD. The significant deficiency of a physiologically important n-3 PUFA, eicosapentaenoic acid (20:5n- 3), in leukocyte membranes in the COPD patients was revealed. In turn, the low level of 20:5n-3 could result from the deficiency of its precursor, docosahexaenoic acid (22:6n-3). The results of the study indicate the modification in the PUFA composition of blood leukocyte membranes in the patients with COPD. It was shown that altered composition of long-chain fatty acid of leukocyte membranes emerges already at the early stage of the disease. Therefore, the imbalance in fatty acids composition of leukocytes makes a significant contribution to the development and the progression of COPD.
The parameters of several populations of immune cells (T cell populations, macrophage subpopulations) in peripheral blood and brain were studied in a clinically significant model of mild traumatic brain injury among rats. The population of resident cells of innate immunity of microglia and brain astrocytes with local tissue damage is involved in the implementation of the inflammatory response, it is also shown that in case of trauma, blood leukocytes can overcome the blood-brain barrier and penetrate the brain parenchyma. The methods of flow cytometry and immunofluorescence were used. An increase in the number of monocytes and neutrophils up to 1 day, after a mild traumatic brain injury (TBI) with a subsequent decrease to the end of the observation period was noticed. It was determined, that the number of CD45+ cells, CD3+T cells decreased at 1 days post-injury (dpi), and rose slightly by 14 dpi, the percentage of CD4+T cells continuously declined from 7 to 14 dpi, while the percentage of CD8+T cells increased from 7 to 14 dpi. With mild traumatic brain injury in animals, a significant (3-10 times) decrease in the number of microvessels with a positive reaction to the presence of SMI 71 on the 8th and 14th day after head injury was observed. Intensive staining of SMI 71 microvessels was sometimes observed with an increase in the area of a positive reaction. Thin positive deposits of the reaction product are observed in the brain of healthy animals around the wall of the microvessel. In the damaged brain, CD45high/CD11b+ positive macrophages of the M1 subpopulation appeared in the brain tissue on the 2nd day after TBI and a significant amount was observed on the 8-14th day. In the corpus callosum and ipsilateral region of the striatum, the content of cells expressing CD16/11b+ reached a maximum 8 days after TBI, which correlated with a decrease in the positive response to the presence of endothelial antigen SMI 71. Thus, in the acute period of mild TBI, the presence of neuroimmunopathological processes is determined in the brain, which can subsequently result to the dysregulation of neuroimmune connections.
In recent years, there has been an increase in the prevalence of allergic diseases in children. Allergic rhinitis (AR) ranks first among other allergic diseases in terms of prevalence and impact on the health and quality of life patients. In various countries of the world, according to various sources, from 10 to 40% of the population suffers. Allergic rhinitis is a serious medical, social and economic problem. In addition, allergic rhinitis, especially with multiple sensitization and insufficiently controlled course, is an independent risk factor for recurrence of respiratory infections and the development of bronchial asthma, and also significantly reduces the quality of life patients.
AR is a multifactorial disease in the development of which many factors play a role. The basis of the pathological process in AR is IgE--dependent mucosal inflammation, which is realized under the influence of specific and nonspecific mechanisms and has a Th2 character. The tissues and organs involved in the process determine the formation of complex mechanisms of interaction between the immune, microcirculatory and autonomic nervous systems.
The inflammatory process in AR is characterized by a number of features, for example, the presence of minimal persistent inflammation and the priming effect, which in turn is a predisposing factor for the clinical onset and progression of AR. Microcirculatory mechanisms are of great pathogenetic significance in the development of allergic inflammation, including in AR.
Depending on the leading pathognomonic trait, it is now customary to distinguish individual phenotypes and endotypes of AR.
The phenotype covers the clinically significant properties of AR, but does not reveal the detailed mechanisms of its development, on the basis of which a personalized algorithm for prevention, treatment and prognosis can be created.
And the autonomic nervous system is responsible for setting links between the body, ambient and internal environment through the regulation of metabolism, functioning of organs and tissues based on changes in this environment; it also provides the integration of all organs into a single whole acting as one of the main body’s adaptive systems.
Since the autonomic nervous system governs the body and homeostasis uniting separate pathogenetic links of disease progression and sets the basis for structural and functional unity. In light of this, the failure of neuroregulatory mechanisms takes the lead among the causes of systemic changes in the microvasculature, which, in turn, reflects general pathogenetic processes in the body. The regulatory mechanism is implemented through nerves and reflexes by different neurohumoral factors, their nature has been studied under experimental conditions and is beyond doubt to date.
The study of the main indicators of microcirculation and the autonomic nervous system among children with allergic rhinitis in various combinations with concomitant pathology will highlight new AR phenotypes and select an individual treatment and rehabilitation plan for these children.
Excessive activity of cytokines, as well as their deficiency, promote a regulatory imbalance of immune system, which may cause persistent inflammation and, as a consequence, loss of pregnancy. The purpose of our work was to study the role of cytokine expression in the development of endometriosis in women exposed to excessive environmental contamination with hydroxybenzene. The study involved 106 women of reproductive age, divided into 4 groups according to two criteria: the presence or absence of reproductive disorders (endometriosis), as well as levels of blood contamination with hydroxybenzene compared to the reference ranges. The groups of women were comparable in age, ethnicity, and social status. The level of phenol in blood was measured by gas chromatography, and the cytokine levels were determined by enzyme immunoassay. The use of parametric and nonparametric statistical criteria as well as adjustments for multiple Bonferoni comparisons, allowed us to establish significant differences between the groups, according to the levels of IL-1β, IL-8, IL-10. The expression of interleukin 8 (IL-8) in women with endometriosis was found to be significantly higher than in healthy ones. At the same time, the observed features of IL-8, interleukin 1β (IL-1β) expression correlate with the overproduction of interleukin 10 (IL-10) associated with excessive contamination of bio-environments with hydroxybenzene. The results of using non-parametric correlation analysis of the Spearman data revealed a positive correlation between IL-10 production and hydroxybenzene levels in the blood, as well as inverse relationship between IL-10 and IL-8 expression. Thus, evidence has been obtained of the involvement of exogenous estrogen hydroxybenzene in development of reproductive disorders, probably, formed under active participation of increased anti-inflammatory IL-10 cytokine, which, by antagonism to proinflammatory mediators (e.g., IL-8), seems to promote apparent transition from acute phase of endometriosis to chronic disorder, thus reducing the reproductive potential of women. It is known that IL-8 expression significantly correlates with development of endometriosis, but excessive phenol exposure may increase the anti-inflammatory IL-10 expression, thereby suppressing the activity of IL-8. Suppressed activity of proinflammatory cytokines by phenol may lead to chronic inflammation and impaired reproductive functions.
Our report concerns the observations made during the treatment of pneumonia with individually selected bacteriophages in HCAI patients on mechanical ventilation. 19 patients on mechanical ventilation whose condition was complicated by antibiotic-resistant pneumonia were examined. The treatment of patients was supplemented with phage therapy, bacteriophages were selected individually for each patient, taking into account the microbial etiology of the disease (Pseudomonas aeruginosa, Кlebsiella pneumoniae, Acinetobacter baumanii). Immunophenotyping of blood lymphocytes was carried out using 2-3-parameter flow cytometry. The functional activity of blood leukocytes was assessed by their ability to produce IFNα and IFNγ during cultivation. The level of interferons production in supernatants collected after cultivation was quantitatively evaluated both by their concentration (ELISA, reagents from “Vector-Best-Europe”, Russia) and by their biological activity. Statistical processing of the results was carried out using the Statistica 6 program according to the nonparametric Mann-Whitney U-test. In the course of successful phage therapy with individually selected bacteriophages overcoming of lymphopenia (if there was one) and an increase in both the number and functional activity of peripheral blood lymphocytes in all patients with pneumonia observed are noted. The relationship between the microbial load (mono- or mixed infection, the number of CFU pathogens of pneumonia, the need for repeated courses of phage therapy) and the degree of deficiency in one or another subpopulation of lymphocytes was not detected. Activation of the immune system achieved after one course of phage therapy was maintained for at least 3 weeks after phage administration was discontinued.
CLINICAL CASES
Primary immunodeficiency is a rare congenital pathology associated with failure of immune system, manifested by disturbances of its functions. These defects lead to increased susceptibility of patients to various infectious agents, as well as the development of autoimmune, malignant and other diseases. Primary immunodeficiency is classified as a rare disease, which was previously associated with a poor prognosis with a high risk of mortality in childhood. To date, the emergence of highly effective treatment methods has changed the course and prognosis of these diseases. Clinicians of various specialties increasingly meet with this pathology in everyday practice, including adult age cohorts. In this regard, early diagnosis of primary immunodeficiency in adults becomes relevant, being associated with choosing optimal therapy, prevention of severe internal organ damage, determination of management strategy for the patient, as well as the need to identify inherited disorders and provide information to the patient’s family. Delayed verification of the diagnosis may cause disability of the patient and development of irreversible, often fatal complications. This article presents our own clinical case with a newly diagnosed clinical condition: Common variable immunodeficiency disorder (CVID), the most common form of primary immunodeficiency in adults. The symptoms of common variable immunodeficiency disorder appear in these patients in adulthood, but a high-quality collected history of the disease will allow you to trace symptoms in the patients even since early childhood. There is a common gap for several years between the onset of the disease and clinical diagnosis, since erroneous diagnosis is often made due to non-specific clinical symptoms that resemble other, more frequent diseases. The prognosis of patients with CVID depends on several factors: frequency of infections, structural disorders in the lungs, the occurrence of autoimmune diseases and the success of infection prevention. Thus, a variety of clinical forms of primary immunodeficiency, lack of awareness of doctors about this pathology, complexity of immunological examination in the general medical network lead to the fact that CVID is not diagnosed for long terms, and patients do not receive the necessary pathogenetic therapy. There is a need for drawing attention of doctors of various disciplines to the fact that the recurrent inflammatory processes of various localization, which are difficult to respond to adequate traditional therapy, may be caused by changes in the immune system, including congenital, genetically determined immunodeficiency.
Alopecia areata is a common inflammatory immune-mediated disorder in which autoimmune response is triggered against hair follicles, thus leading to non-scarring hair loss on the scalp, face and other parts of the skin. Despite numerous studies concerning this issue, today there is no consensus on the etiology and pathogenesis of focal alopecia. In the literature, special attention is paid to association of focal alopecia with autoimmune diseases, such as rheumatoid arthritis, celiac disease, type 1 diabetes, psoriasis, autoimmune thyroiditis, vitiligo. Recent studies have identified the association of focal alopecia with atopic diseases (allergic rhinitis, bronchial asthma, atopic dermatitis) and the early debut of severe forms of hair loss. The aim of this study was to present a clinical case of focal alopecia in an 8-year-old girl with atopic bronchial asthma and seasonal allergic rhinitis. As based on detection of eosinophilia in peripheral blood and a high concentration of total IgE in serum, one may assume that atopic alopecia is the cause of focal hair losses in a child with atopy. The patient underwent skin prick testing, in order to determine sensitization for food components, pollen and fungal allergens. As a result of skin testing, a hyperergic reaction (> 15 mm in diameter) to tree pollen was revealed, a positive response (6-9 mm) to oatmeal, a weakly positive reaction (3-5 mm) to whole chicken egg, carrots, tomato, apple, pear, pollen of meadow, cereal, weed grasses was also revealed. With regard of these allergological data, an individual diet was recommended with the elimination of causally significant allergens (including those eliciting weakly positive reactions), external treatment, i.e., topical calcineurin inhibitors administered for 1 month. One month later, an improvement of the pathological process was registered, and 6 months from the start of therapy, complete restoration of hair follicles was noted in the focus of alopecia. The patient was monitored for a year, no complaints of hair loss were noted. The positive effect of elimination against the background of the appropriate elimination diet with respect to causally significant allergens, was also noted when treating her for respiratory allergy, i.e., the patient did not have seasonal manifestations of hay fever over the next pollination period. This clinical case is demonstrated in order to draw special attention of dermatologists, allergologists, immunologists, general practitioners to the issues of focal alopecia in children against the background of typical allergic diseases.
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