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Non-inflammatory exudative lesions of Reinke’ pos space present the most common cause of phonation disorders in representatives of vocal professions. The leading role in pathogenesis of this disorder is given to the nearly complete absence of lymphatic drainage of Reinke space and local deposition of tissue fluid. The mechanisms of its progression are of particular importance when determining the condition of mucosaassociated laryngeal tissue. The aim of our work was to evaluate the parameters of immunohistochemical lymphocyte typing in exudative lesions from the Reinke’s space. Materials and methods: The main group consisted of 40 patients, at the mean age of 43.2±2.1 years, exhibiting tumor-like exudative lesions of Reinke’s space. The biopsy material was taken from the vocal folds, including polyps, vocal nodules and Reinke’s edema. Myxoid and angiomatous types of the polyps were separately assessed. Videofibrolaryngoscopy was performed using an Olympus TYPE 150 bronchofibroscope (Germany). Morphological studies were carried out using a DMRXA microscope (Leika, Germany) by means of the ImageScopeM computer program (Germany). The uniformly treated sections were stained with Hematoxylin & Eosin (Biovitrum, Russia). Immunohistochemical quantitative assessment of the main T cell populations (CD3+, CD4+, CD8+), B cells (CD20+), histiocytes (CD68+), and the cells expressing bcl-2 and p53 cell regulators was carried out automatically using the BenchMarkXT immunohistotainer (Ventana, USA). The results were expressed as U/mm2. Results and Discussion. Some special features of cellular immunophenotype were revealed in exudative lesions of Reinke’s space. Reinke’s edema was characterized by high content of CD3+ lymphocytes, CD4+, CD8+, CD20+, p53 positive cells of the basal epithelium, as well as low numbers of CD68+, bcl-2 positive lymphocytes and cells of the basal epithelium. Myxoid type of polyps was characterized by low content of CD3+, CD4+, CD8+, CD20+ lymphocytes, bcl-2 positive lymphocytes and basal epithelium cells, CD68+ monocyte-macrophage cells, and high amounts of p53 positive basal epithelial cells. In the angioma-type polyps, we have registered low contents of CD3+, CD4+, CD8+, CD20+ lymphocytes, high numbers of monocyte-macrophage CD68+ cells, MMP-9+, bcl-2 positive lymphocytes, and low content of p53 positive cells of basal epithelium. The samples from the vocal nodules were characterized by low content of CD3+, CD4+, CD8+, CD20+ and p53 positive basal epithelial cells; high numbers of CD68+ cells (monocyte-macrophage series), MMP-9+ and bcl-2 positive lymphocytes.

Chronic rhinosinusitis with nasal polyps (CRSP) is a heterogenous disease. We have earlier detected differences in severity of clinical manifestations, cellular infiltration degree shown in nasal polyps, of eosinophil-to-neutrophil ratio, efficacy of intranasal glucocorticosteroid baseline therapy, and various inflammatory patterns for several cytokines on the mRNA expression level in different phenotypes with isolated CRSP cases, CRSP associated with respiratory allergy (RA), or non-allergic bronchial asthma.The purpose of this work was to study the cytokines of TGF-â family in the tissues of nasal polyps in patients with different CRSP phenotypes. The research involved 292 patients suffering from CRSP divided into 3 phenotypic groups. Group I consisted of patients with isolated CRSP free of associated BA and/or sensitization to atopic allergens. Group II included patients with CRSP combined with respiratory allergy and was further divided into two subgroups. I.e., Group 2a comprised patients with CRSP, allergic BA (aBA), and allergic rhinitis (AR), while the patients with CRSP, AR, and non-allergic BA were placed to the group 2b. The patients suffering from CRSP complicated with non-allergic BA were allocated to the group III. The patients with hypertrophic rhinitis served as control. The levels of TGF-â1, TGF-â2, and TGF-â3 proteins (pg/mg) were measured by means of multiplex immunoassay approach in supernates of tissue homogenates from nasal polyps removed by surgery, and in posterior parts of inferior nasal conchae. The total protein level was determined in tissue supernatant, with cytokine contents recalculated for the mg/ml protein concentration for standardization of measurements.In the control group, trace concentrations of all three growth factors were detected. Significant difference in protein contents was found for the studied cytokines, depending on CRSP phenotype. The levels of TGF-â1 and TGF-â2 were statistically lower in isolated CRSP than in other groups of comorbid CRSP patients. TGF-â1 and TGF-â2 concentrations were significantly lower in CRSP + allergic BA group IIa than in CRSP + nonallergic BA and CRSP + RA groups. The amount of TGF-â3 cytokine was maximal in CRSP + non-allergic BA group III compared to the patients with isolated CRSP of group I and CRSP + non-allergic BA group 2a.Conclusions.The high level of all three TGF-â isoforms in patients with CRSP compared to the control group suggested a high potential of mucous membranes of paranasal sinuses for active tissue remodeling followed by nasal polype formation.Different mechanisms were presumed for development of local pathological process in different clinical phenotypes of CRSP, depending on the comorbid pathology, especially, BA or respiratory disorders.Minimal TGF-â1 and TGF-â2 levels were detected in isolated CRSP.The highest concentrations of TGF-â1, TGF-â2, and TGF-â3 were discovered in the patients with CRSP accompanied by non-allergic BA as compared to the groups with isolated CRSP and CRSP+allergic BA.5. Determination of TGF-â1, TGF-â2, and TGF-â3 levels can serve as an additional criterion for differentiating between the mechanisms of mucous membrane damage in local pathological process in tissues of comorbid patients with different CRSP phenotypes.

Granulocyte-macrophage colony-stimulating factor (GM-CSF) belongs to the group of growth cytokines (hematopoietins) that regulate proliferation and differentiation of myeloid lineage cells. Recently, a lot of new data have accumulated, indicating the presence of a number of previously unknown biological effects in GM-CSF and synthetic peptides of its active center, which open up new prospects for their wide clinical use.The review outlines current understanding of the structure, functions, and mechanisms of GM-CSF action and concerns the structure of its receptor. The GM-CSF producer cells are characterized, as well as target cells (effector cells) responding to this cytokine are also presented. The known mechanisms of intracellular signaling involved into the GM-CSF/receptor interaction are described. The main pleiotropic effects of this cytokine as a factor of hematopoiesis and an immunostimulating agent are characterized. The previously known and recently found immunobiological effects of this cytokine, its recombinant forms and synthetic analogues of its active center are discussed.Participation of GM-CSF in hematopoiesis and differentiation of myeloid cells, the effects of this cytokine on the functional activity of immunocompetent populations (lymphocytes, macrophages, neutrophils, dendritic cells) and tissue cells were characterized. The influence of GM-CSF on the development and course of infectious and inflammatory processes, its role in the creation of combined vaccines is reviewed. Clinical data on usage of GM-CSF and its recombinant forms in hematology, immunology, oncology, reproductive medicine and in the treatment of systemic autoimmune processes and infectious diseases are presented.The recently discovered immunobiological properties of synthetic peptides derived from active center of GM-CSF are summarized, indicating that they exhibit immunotropic and hematopoietic effects, as well as antimicrobial activity against Gram-negative and Gram-positive bacteria, viruses, and tissue repair (effect on the rate of wound healing), which is not typical to the whole GM-CSF molecule. We discuss the prospects for clinical applications of synthetic GM-CSF analogue (ZP2 peptide), and an opportunity of creating new cosmetics and pharmaceuticals with combined immunostimulating, antimicrobial and reparative properties on its basis.The review expands the view on potential usage of cytokine therapy in the treatment of various infectious and non-infectious diseases in humans, and is addressing a wide range of specialists working in the field of allergology and immunology, infectology and regenerative medicine.

Treatment of chronic viral infections accompanied by permanent virus persistence in the target epitopes of the oral cavity, skin, urogenital tract is complicated by virtual lack of available drugs exerting combined systemic virulicidal and immunomodulatory effects. Here we demonstrate clinical and immunological efficacy of combined therapy in treatment of Epstein–Barr virus (EBV)-associated chronic infections. The aim of the study was to evaluate the clinical and immunological efficacy of combined etiopathogenetic therapy using the Acegram cosmetic product in patients with EBV-associated chronic infections. Materials and methods. There were enrolled 40 patients monitored before treatment as well as 20 patients followed up after combination therapy (cycle therapy consisted of oral valaciclovir (Valtrex) applied at dose of 500 μg twice a day for 10 days, glucosaminylmuramyldipeptide (Licopid) — 10 mg 2 twice a day for 10 days, topical irrigation for mucous membranes with granulocyte-macrophage colony-stimulating factor active center-derived peptide (Acegram-spray) 3 times a day for 10 days. If necessary, treatment courses were repeated 20 days after the onset. All patients were examined for the presence of EBV genomes in the oral fluid and blood using the qualitative and quantitative polymerase chain reaction (PCR) using the DNA technology test system (Russia) on a DT-Lite device prior treatment and 30, 60 days post-therapy time points. In addition, serum samples were analyzed for level of class G immunoglobulins specific to the EBV nuclear and capsid antigens by using enzyme immunoassay (test systems manufactured by CJSC Vector Best, Russia) as well as immune status (clinical methods, enu flow cytometry evaluation of the phagocytic activity of neutrophils, ELISA method). Results. Use of single or two course combination therapy in subjects with fully eradicated EBV carriage associated with reversed clinical symptoms was accompanied by recovered immune system status (T and B cells, T-helper cells, CD3+ CD25+ cells, phagocytosis parameters). A non-invasive approach proposed for controlling virus elimination in the oral fluid by using polymerase chain reaction method may serve as to objectively monitor therapeutic efficacy.

Antibacterial drugs are routinely used in therapy of periodontal diseases. However, increasing incidence of antibiotics resistance necessitates development of novel therapeutic approaches for oral diseases. Currently, newly designed antibacterial agents based on natural, semi-synthetic and synthetic peptides is the most promising approach in dentistry. Among them is a product containing synthetic peptide (ZP2) replicating active site of granulocyte-macrophage colonystimulating factor (GM-CSF) as the main active ingredient in Atsegram-spray (manufactured by Academic Innovation Research Center, Chelyabinsk). Our study was aimed at assessing efficacy of a combination therapy of inflammatory periodontal diseases by introducing Atsegram-spray as well as examining potential relationship between peptide ZP2-related biological properties, physicochemical properties of the spray and mechanism of antibacterial and immunotropic action for substantiating its application on oral mucosa. During the first stage experiments, it was found that the peptide ZP2 was able to trigger lymphocyte blast transformation in vitro indicating that it might influence cell proliferation and exhibit marked immunotropic activity. Next, we assessed potential effects of the peptide ZP2 on biofilm formation mediated by staphylococcal clinical isolates. It was shown that peptide ZP2 inhibited biofilm formation in 75.0±9.0% of S. aureus and 50.0±15.1% of S. epidermidis strains, with mean inhibition index of biofilm formation reaching 25.1±3.8 and 50.4±6.0%, respectively. However, peptide ZP2 in 8.3–25.0% of staphylococcal clinical isolates was found to stimulate/lack effect on biofilm formation by 14.9–48.5 and 16.7–25.0% cultures, respectively. Thus, the synthetic peptide ZP2 exerts divergent, but mainly inhibitory effects on biofilm formation with staphylococcal clinical strains, which are characterized by inter- and intraspecific (strain) variability. Use of a synthetic peptide-based spray in antibacterial therapy of mild chronic generalized periodontitis (main group) one month after the onset was found to improve oral hygiene by 28.5% as well as decrease PMA index and gum bleeding index by 82.8 and 100%, respectively. In contrast, such parameters in patients receiving basic therapy (comparison group) were lower on average by 2-fold. While analyzing physicochemical properties of the spray such as pH, buffer capacity and solution osmotic pressure, it was found that they were related to the antibacterial mechanism of drug activity and efficacy in treatment of inflammatory periodontal diseases. Thus, assessing peptide ZP2-related biological properties and physicochemical parameters of the spray allows to evaluate their role in mechanism of previously unknown antibacterial and immunotropic activity. These findings confirm feasibility and efficacy of using Atsegram-spray in dentistry as an alternative means to antimicrobial agents, such as antibacterial drugs.

Despite numerous attempts to control the course of chronic rhinosinusitis with nasal polyps (CRSwNP) by means of pharmacological treatment and new surgical approaches, the majority of patients experience lifelong persistence of this disorder, at recurrence rates of 50-60% within 18 months after surgical treatment. Since CRSwNP is a chronic persistent inflammatory process, it affects the entire body condition, including the state of systemic immune response. An elevation of NK (CD3-CD16+CD56+), activated NK (CD8+CD3-), NKT cells (CD16+CD56+CD3+), Treg (CD4+CD25brightCD127low to neg) cells and activated T-lymphocytes (CD3+CD25+) was revealed elsewhere among all the patients with CRSwNP, using a flow ytometry method. There was no difference between various disease phenotypes. We analyzed the status of cellular component of systemic immunity, dependent on clinical course of the disease and efficiency of the administered therapy of CRSwNP. The patients were divided into three subgroups. The follow-up period was 1 year. The first group comprised the patients who showed positive dynamics after conservative therapy, resulting into regression of nasal polyps and their grade than a year ago. The second group included the patients in whom the size of polyps remained the same. The third group included the patients with higher incidence of nasal polyps than a year ago.We have shown a decrease of Treg, NKT cells, NK and activated NK, cytotoxic T-lymphocytes (CD3+CD8+), activated T-cell numbers in clinical group 3 with aggressive growth of polyps and low effect of standard therapy, which may cause deterioration of the immune system cellular populations, accompanied by presence of persistent productive inflammatory process of nasal cavity and paranasal sinuses. In the second group, a significant elevation of total lymphocyte number, total and activated T cells, T helpers (CD3+CD4+), cytotoxic T lymphocytes, NK and NKT cells was shown. Meanwhile, a decrease in absolute number of activated NK was observed despite the NK growth. Therefore, we can assume that the mechanism of their activation was disturbed and compensated by production of NKT cells and cytotoxic T lymphocytes. Moreover, we have shown in this group that the absolute number of Treg cells is increased; and these cells had a suppressive influence on effector cells of adaptive immune response, thus inducing incomplete elimination of infectious agents, which contribute to permanent incomplete course of inflammatory process. Chronic inflammatory process in CRSwNP affects systemic cellular immunity depending on the morbidity characteristics in the course of pathological process. The maximal intensity of systemic cellular immunity is observed in the group of patients that require permanent basic drug therapy. In case of aggressive CRSwNP and failure of standard drug therapy, we observed a decrease in absolute numbers of effector cells, along with decreased Treg lymphocyte numbers which may explain inefficient immune regulation of inflammatory process and medical interventions in this group of patients.

We studied clinical efficacy of sublingual immunotherapy in children with combined respiratory forms of pollen allergy before and after three courses of treatment. The levels of G-CSF, GM-CSF, IL-10, IL-12p70, IFNγ, IL-13, IL-17A, IL-1β, IL-2, IL-4, IL-5, IL-6, IL-7, TNFα, IL-8, MCP-1, MIP-1β were measured in nasal secretions. In remission state, the children with combined forms of respiratory allergy to pollen allergens showed local signs of chronic inflammatory process (in the nasal mucosa) associated with disturbed immune response, with increased synthesis of proinflammatory cytokines and decreased production anti-inflammatory cytokines, as well as enhanced reactions aimed for allergen elimination. Following clinical success of sublingual immunotherapy, we have shown a reduction in immunological imbalance, due to decreased activity of the systems modulating severity of inflammation and production of pro-inflammatory cytokines.

Urban ecological problems are associated with industrial pollution emissions, waste of metallurgical production, unhealthy lifestyle and reproductive behavior lead to a steady rise in cancer pathology in theChelyabinskregion. Women in early reproductive age living in areas of ecological concern with suspected mastopathies, exhibit significantly higher levels of tumor markers associated with the tumor cell membranes of (CA19-9, SA15-5, CA125), CEA (carcinoembryonic antigen). Therefore, it seems necessary to perform massive laboratory programs for oncological screening, especially, among young working population, in order to identify breast diseases, and to arrange appropriate risk groups at this basis, aiming for development of effective preventive measures for the future.

Several endogenic and synthetic peptides were investigated in order to reveal their antibacterial activity. The study of dose-depended effects of antibacterial activity of the immunotropic peptide ZP-2, which is included in GM-CSF active center, shown significant antibacterial activity of ZP-2 by action range and by rate of bactericidal activity in case of low concentration. Combined application of substance derived from supernatant of CD34+CD45-cells and synthetic peptide ZP-2 enhances significantly actions of each other. Probably, this effect is connected with various mechanisms of their influence on bacteria. The obtained facts indicate that synthetic peptide has not only immunostimulating action but it also promotes a direct antibacterial activity. These data support an idea that synthetic peptides have good perspectives in case of development new antibacterial drugs with unique characteristics.

T-lymphocytes play an important role in elimination of tumor cells, in reactions of a transplant against graft and graft versus host disease, in slow-type hypersensitivity, and other reactions directed for maintenance of homeostasis. Along with CD3, an antigen-specific T-cellular receptor (TCR) is another common marker of T-cells. There are two types of TcR – αβ-TcR and γδ-TcR that differ in ontogenetic and functional properties. γδ-T-cells play a significant role in protection of organism against various types of infections, and determination of their amounts should be an integral part of the analysis of patients’ immune status. To these purposes, a multi-colour analysis shuld be used, applying the following combinations of monoclonal antibodies: CD3/CD4/CD8/CD45 and αβ-TcR/γδ-TcR/CD3/CD45. Multi-colour staining and multi-step gating allow of carrying out multiparametric analysis of peripheral blood with high accuracy and reliability. The proposed approach considerably facilitates interpretation of results obtained, and it allows of judging about immune system functioning in various pathological conditions.

Over last years, huge efforts are made in order to improve analytic techniques for T lymphocyte subpopulations. Appropriate studies on T-helper cells are also subject to extensive research. These cellular compartments are substantially divided into various sub-classes. Such subpopulations, e.g., Th1, Th2, T-reg, Th17 and activated Т-helpers, were classified and characterized to sufficient degree. Recently, a lot of distinct data about specific receptors and functional properties of various Т-helper subpopulations was obtained. Applications of these results in laboratory research, without any doubt, will improve diagnostic quality when assessing functional alterations of immune system, as well as adequate therapy planning. ( Med. Immunol., 2011, vol. 13, N 1, pp 7-16)

Аbstract . The objective of present study was to determine the immunological features of immune system aging in mucosa-associated lymphoid tissue (MALT) in the patients at different ages (mature, aging and old) observed at a dental unit. A study of cellular spectrum and humoral factors in salivary gland secretions has been performed in a group of 106 persons (35 to 90 years old). A number of age-dependent features of the immune profile were revealed for the mucous-salivary area, thus characterizing involution events within MALT structures. Among specific markers determining intensity of MALT-associated aging, a decreased percentage of viable immune cells (below 40%), along with the prevalence of the neutrophilic granulocytes in the salivary secretions (over 98%) (with increased expression of β2-integrins); decreased counts of mononuclear cells, i.e., mononuclear cells with low expression of CD11β adhesion molecules, B-lymhocytes, and Th-lymphocytes have been revealed. Alterations in serum factors included a general decrease in complement system activity (СН50) and anaphylotoxines (С3а,С5а); elevated protein, mucine, and IgМ levels. The revealed specific features of cellular and humoral immunity within MALT-associated muco-salivary zone may be considered as a normal response connected with natural aging processes. (Med. Immunol., 2011, vol. 13, N 2-3, pp 167-174 )

We analyzed efficiency of application for analytic control parameters, i.e., «T-cell replicates» and «T-cell summary» in a sample of 174 HIV-infected patients, independently of age or disease stage. The patients were divided in two groups. Group 1 consisted of patients with total CD45+CD3+CD4+ and CD45+CD3+CD8+ lymphocyte concentrations being similar to relative and absolute concentrations of CD45+CD3+ lymphocytes, or the difference between absolute measurements not exceeding «T-cell replicates» differences. Group 1 included 36.78% of the whole cohort. Group 2 consisted of patients with total CD45+CD3+CD4+ & CD45+CD3+CD8+ lymphocytes concentrations being lower than relative and absolute concentrations of CD45+CD3+ lymphocytes, and the differences in absolute values being higher than «T-cell replicates» differences (41.38% of total sample). Such difference between control sums may be connected with high concentrations of double-negative γδ-T-cells (CD45+CD3+CD4–CD8–) in the specimens under study. (Med. Immunol., 2008, vol. 10, N 4-5, pp 463-466) .

Incidence of different types of dislipoproteinemias (DLPs) and a balance between apoptosis and proliferation of lymphocytes were studied in males exhibiting initial manifestations of cerebral blood flow insufficiency (ICBI). In general, high intensity of lymphocyte activation was revealed in patients with ICBI, with amplified proliferative potential demonstrable in this group. We have also shown a trend to increased numbers of T and B lymphocytes expressing antiapoptogenic bcl-2 protein. The changes in apoptosis and cell proliferation intensity were more pronounced in IIA and IIB types of DLP. In these groups, unidirectional changes of FasR expression were observed. However, the parameters reflecting readiness to apoptosis were changed multidirectionally. Cholesterol and oxysterols transported to the cells under excessive dyslipidemia may influence activities of transcription factor genes (bcl-2, Fas-regulatory gene (TDAG51), thus, probably, causing an imbalance between cellular proliferation and apoptosis.

Flow cytometry is considered as a modern technology for fast measurements of cellular characteristics, their organelles, and processes occurring within them. It is regarded as an efficient solution in many important areas of cell biology, immunology and cellular engineering. Present article bears on main developments in flow cytometry and their applications in medical and biological practice. Usage of modern achievements in fluorescent dyes, progress in laser and computer technologies, as well as potent software, resulted in wide application of this technique in medical practice. Accordingly, usage of monoclonal antibodies conjugated to different fluorochromes has led to elaboration of multiparametric analysis and did sufficiently simplify specialized works aimed for diagnostics of various immune disorders. The new directions in flow cytometry, e.g., flow cytoenzymology, provide wide opportunities for detailed identification of damaged or altered cells, and taking adequate decisions in treatment of detected pathological changes. The authors suggest that this article could initiate a series of publications concerning usage of this technology and its modern applications in broad laboratory practice.

Development of modern clinical and infectious immunology, demands maximum objective tests for estimation of functional condition of various types phagocytic cells, such as neutrophils, monocytes, macrophages and dendritic cells. Wide potential opportunities of flow cytometry have led to development of ways of registration of a functional condition of these cells, an estimation of processes of their activation and study of mechanisms formation of defects of their functions. For example: analysis of phagocytic and bactericidal activity of neutrophils and measurement number of the cells, which are taking place at various stages of programmed cellular death (apoptosis). Flow cytometry methods described in this review represent a new hi-tech direction for scientific and clinical researches.

Flow cytometry allows estimating quantitative and qualitative structure of populations and subpopulations of immune system cells by using various methodical approaches and a wide spectrum of reagents. For diagnostics the Acquired Immune Deficiency Syndrome (AIDS) caused by a Human Immunodeficiency Virus (HIV) the flow cytometry became irreplaceable. Traditionally, immunologists examine standard model of an estimation of immune dysfunction on the basis of classical markers of Т-cells (CD3, CD4, CD8) at the HIV-infection. But researchers pay less attention to other populations and subpopulations of lymphocytes, such as γδ-, αβ- and CD38+ Т-cells. The quantitative estimation of these parameters from a HIV and AIDS patients enables to see pathogenesis a HIV infection and the prediction of its development from another side.

Development of a modern medical science demands more thorough and deeper approach to diagnostics of diseases. New diagnostic technologies for these investigations are necessary and flow cytometry is such technology. Modern achievements in the field of fluorescent dyes, development of laser and computer technologies, have led to wide use of the given technology in medical practice. Infectious immunology is not exception. Cytokines production analysis at tuberculosis, HLA-DR expression changes for monocytes and CD64 for granulocytes at sepsis are the important diagnostic attributes. Flow cytometry offers new approaches to early and faster diagnostics of infringements of immune system at tuberculosis and sepsis, allows supervising development of these pathologies more effectively and estimate efficiency of therapy.

Patterns of immune response, cytokine- and NO-ergic mechanisms and their role in the development of basal-cell skin cancer were evaluated in population of an industrialized region. Multiple forms of basal-cell skin cancer and primary multiple cancer were characterized by the evidence of Th2-dependent imbalance of immune response, decreased cytolytic potential, increase of EGF, TGF-β, IL-1β, IL-4, IFNγ, lactoferrin levels, thus suggesting a particular immune defense strategy in malignant growth.

This review is focused on analysis of currently used flow cytometric methods designed for identifying apoptotic cells in various invertebrate and vertebrate species. Apoptosis can be characterized by stage-specific morphological and biochemical changes that are typical to all kinds of eukaryotic cells. In this article, we consider different techniques of apoptosis detection based on assessment of cellular morphology and plasma membrane alterations, activation of intracellular enzymes and components of a caspase cascade, as well as DNA fragmentation and failure of mitochondrial transmembrane potential, as assessed in various animal groups. Apoptosis recognized as a key mechanism aiming at maintenance of cellular homeostasis in multicellular organisms, and such investigations represent a necessary component of fundamental and applied studies in diverse fields of experimental biology and immunology. A broad spectrum of apoptosis markers is used, and the preference is given to optimal approaches, as determined by experimental tasks, and technical opportunities of the laboratory.

We have performed immunophenotyping of lymphocytes in 150 HIV-positive patients, by means of flow cytometry. All the persons under study were classified into three groups, depending on absolute contents of CD3+CD4+T-lymphocytes in peripheral blood, according to HIV infection staging in adults and adolescents, as amended by CDC (USA). It was revealed that, at each subsequent stage of disease, a decrease in absolute numbers of T-helpers is accompanied by drop in relative and absolute numbers of lymphocytes, as well as in total T-lymphocyte numbers, and decreased amounts of absolute cytotoxic T-lymphocytes levels, combined with increase in their relative contents, along with decreased CD3+CD4+/CD3+CD8+ ratio, diminished levels of double-positive subpopulation, decrease in relative and absolute levels of αβТ-lymphocytes and significant drop in absolute levels of γδТ-cells, while their relative amounts remained normal. These findings demonstrate a pattern typical to the absence of immunological reactions in response to antigens persisting in the organism.

We have performed immunophenotyping of lymphocytes, employing a flow cytometry approach, along with evaluation of spontaneous or mitogen-induced production of cytokines (IFNγ, IL-4, IL-1β, IL - 10), in the patients with atopic and bacterial (mixed-type) bronchial asthma. We have detected mostly similar changes in lymphocyte parameters. However, among the patients with bacterial asthma, an imbalance of immune cells was more pronounced. Some common alterations of cytokine production were found, i.e., a decrease in IFNγ and IL-1β production, along with increased IL-4 and IL-10. These changes were more significant in the subjects being in acute phase of either atopic, or bacterial asthma. Moreover, IL-10 suppressed IFNγ to higher degree, than IL-4, thus favoring distinct predomination of Th2-type immune response.

Determination of lymphocyte subpopulations and their phenotypes is an important diagnostic feature, in order to elucidate some disturbances connected with immune system functioning. However, insufficient data are obtained when analyzing only major populations of peripheral lymphocytes. In order to perform clinical diagnostics, the data about minor lymphocytic populations and activated cellular pools seem to be more pertinent. Studies of peripheral blood cell subpopulations of healthy donors performed in different Russian regions allowed to assess quantitative distribution intervals for both major and minor immune cell subpopulations in humans. The results obtained, as compared with data from literature, provide an evidence for similar reference intervals for main immune cell subpopulations in healthy donors, independent on their habitation area. Present work has resulted into development of algorithms for cytometric studies and generation of certain panels of monoclonal antibodies enabling evaluation of all main lymphocyte subpopulations, as well as their minor subsets participating in emerging immune response. The distribution intervals have been estimated for such minor subpopulations, as B1- and B2-lymphocytes, memory B-cells, γδ- and αβT-cells, regulatory and naїve T-cells, cytotoxic and secretory NK-cell polupations. The results of present study, while been performed with peripheral blood of healthy donors, may provide a basis of reference values when studying subpopulation profile of immune cells.

A study of leukocyte regulation system in patients with exacerbated bronchial asthma (BA) was performed by means of measuring spontaneous and induced production of certain cytokines (IL-4, IL-1b, IL-10 and IFNg) in whole blood, and drawing some specific coefficients reflecting production of the cytokines, as compared to control values. Group 1 consisted of mixed-type BA patients, with mild clinical manifestations at exacerbation stage. A control group included twelve healthy persons. The cytokine network was evaluated according to spontaneous production of cytokines (IL-4, IL-1b, IL-10 and IFNg), or their induction with standard mitogens (PHA and E. coli LPS). The results obtained have shown that in BA as a clinical model of chronic inflammation, synthesis and activation of the cytokines depend on the phase of disease. At stage of exacerbation, an imbalance occurs between pro- and anti-inflammatory cytokines, with predominance of anti-inflammatory IL-10, which is targeted to suppression of bronchial inflammation.

The article considers an original approach to individual dose selection and efficiency evaluation of immunity-targeted drugs applied for treatment of children with trauma, based on examination of phagocyte locomotive functions.

Temporary immunodeficiency is often associated with various pathological conditions, such as myocardial infarction, severe trauma, or major surgery. However, there are no clinical criterions indicative for presence or absence of such immune deficiency. Meanwhile, clinical signs of infectious complications may be absent because of deficient immune response. A technique for monitoring septic conditions in the patients with open traumas and during postsurgical period has been proposed, employing a flow-cytometric approach. The evaluation principle for septic conditions consists of measuring expression of HLA-DR antigens at the surface of peripheral blood monocytes. As a criterion of patient evaluation in septic state, a relative amount of monocytes expressing HLA-DR may be applied, and the disease prognosis is considered as favorable, if the amounts of positive cells exceed 40% by day 5 after the patient was admitted to the hospital, and an adequate treatment was carried out. This technique may find wide application for estimation and monitoring of septic conditions in the patients. (Med. Immunol., vol. 10, N 4-5, pp 379-388).