Blast Transformation Reaction Research Articles

Некоторые особенности токсикологических свойств специфического иммунобиостимулятора «Трансфер-фактор» в доклинических испытаниях

Abstract. The purpose of the study is to evaluate some aspects of toxicological safety of a specific immunobiostimulant “Transfer factor” in laboratory animal models. Methods. The experiments were performed on mice, rats and guinea pigs. The toxicological safety assessment of the “Transfer factor” preparation included the determination of the following characteristics: chronic toxicity, assessment of specific activity, assessment of embryotoxic and teratogenic properties, assessment of allergenic properties Results. It was established that the introduction of the “Transfer factor” preparation in a chronic toxicological experiment is accompanied by the development of muscle tremor in the rodents’ body, the duration of which depends on the administered dose, route of administration and exposure time, but their body weight increases by 6.29–10.63 %. Autopsy of experimental group animals revealed no visible changes in the arrangement of internal organs and fluid accumulation in the abdominal and pleural cavities, although some pathological changes in color, consistency and size of the lungs, spleen, liver and heart were noted. The mass coefficients of these organs change with an increase in the dose of the administered drug “Transfer factor”, especially with the intraperitoneal route of administration up to 10.39 %. The tested drug in the reaction of lymphocyte blast transformation increases the number of blasts from 0.20 to 1.40 %. The totality of data allows us to state that the drug “Transfer factor” in accordance with GOST 12.1.007-76 belongs to the IV hazard class “low-hazard substances” and it can be recommended for further clinical trials, in which a dosage exceeding 6 ml/kg of live weight will not be used, when administered to laboratory animals, a complex of changes in the internal organs develops. Scientific novelty. The introduction of “Transfer factor” does not affect the condition and functions of the reproductive organs (uterus, ovaries) of pregnant rats, and the drug does not exhibit a negative embryotoxic and teratogenic effect in their body. When studying the allergenic properties of the drug, it was found that it does not cause a general anaphylaxis reaction in the body of guinea pigs, does not irritate the skin in the reaction of immune complexes and the conjunctiva of the eye in the conjunctival test.

INTEGRATED ANALYSIS OF IMMUNOLOGICAL BLOOD PARAMETERS IN PRESCHOOL CHILDREN WITH RECURRENT RESPIRATORY INFECTIONS

Background. Recurrent respiratory infections are a very pressing problem in everyday pediatric practice. They are most commonly diagnosed in preschool children. According to different researchers, their incidence varies considerably (6-30%). The temporary immaturity of immune system is thought to be one of the causes of frequent episodes of acute respiratory infections (ARI). Aim: To determine the state of interdependence between the particular indicators of complex immunological blood test and the frequency of ARI episodes in preschool children. Materials and methods. A total of 26 children (11 boys and 15 girls) aged 1-4 years, undergoing inpatient treatment on ARI, were involved in the clinical study. The number of ARI episodes during the previous year of their life was taken into account. Additionally, two integral indices of ARI recurrence, specifically the infection index and resistance index, were calculated. The children underwent a clinical blood test. Also, the particular indicators of humoral and cellular immunity, functional activity of immune cells (neutrophil phagocytic activity (PhA), lymphocyte blast transformation reaction), complement system (C3c, C4) and the several classes of circulating immune complexes (CIC) were analyzed. The statistical processing of the digital data was performed using IBM SPSS Statistics 28 licensed software with the calculation of the confidence interval (CI) for the main indicators. Results. The prevalent majority of the examined children (80.8%) had an increased serum CIC(small) level. There was a moderate positive correlation between the resistance index values and the CIC(small) level [ρ = 0.392; p = 0.048; CI: 0.072–0.644]. The tightest interdependence was found in the following pairs of studied indicators: 1) CIC(medium)*CIC(small) [ρ=0.732; p<0.001; CI: 0.471–0.875]; 2) C3c*C4 [ρ=0.658; p<0.001; CI: 0.352–0.837]; 3) CIC(large)*Ig(A+M+G) [ρ=0.643; p<0.001; CI: 0.330–0.829]; 4) PhA(spontaneous)*PhA(induced) [ρ=0.601; p=0.001; CI: 0.267–0.806]. A linear regression analysis showed that the predicted resistance index was dependent on only one of the immunological blood parameters considered, specifically total leukocyte antigen. Among other possible predictors, the impact of this antigen on the dependent variable was 18.4%. Conclusions. The particular indicators of a complex immunological blood test are highly interrelated. The results obtained should be taken into account when developing an integrated model for predicting the likelihood of recurrent ARI episodes in preschool children.

Analysis of mononuclear leukocyte functional activity in peripheral blood of patients with varying clinical severity of acute Lyme disease

The aim of the study is to assess spontaneous and phytohemagglutinin (PHA)-induced proliferative and cytokine secretory activity in mononuclear blood cell cultures of patients with acute Lyme borreliosis (LB) over times before and after antibiotic therapy as well as depending on clinical severity. Materials and methods. Activity of blast transformation reaction (BTR) of lymphocytes and the secretion levels of interleukins (IL)-1β, IL-2, IL-6, IL-4, IL-10, IL-8, tumor necrosis factor (TNF)-α, monocyte chemoattractant protein (MCP)-1, interferon (IFN)-γ and IFN-α were assessed twice over time in the cultures of mononuclear blood cells of 18 and 14 patients with mild and moderate severity of acute LB. Results. It was determined that the moderate clinical course of acute LB as compared with the mild course was associated with the significant suppression of BTR levels, the increases in spontaneous and/or PHA-stimulated production levels of proinflammatory cytokines IL-1β, IL-6, IL-8, TNF-α and MCP-1, which were balanced by the increases in secretion levels of anti-inflammatory cytokines IL-10 and IL-4. The initial suppression of IFN-γ secretion in this group gave way to an increase in its production over time, which generally corresponded to the formation of an immune response of the mixed Th1/Th2 type.

INFLUENCE OF MODIFIED PEPTIDES FROM THE FETAL THYMUS ON THE ACTIVITY OF T-LYMPHOCYTES AND NATURAL KILLERS IN EXPERIMENTAL VIRAL HEPATITIS

The results showed that immunomodulin has a moderate regulatory effect on the spontaneous blast transformation of lymphocytes in the in vitro system. The costimulatory effect of immunomodulin in the reaction of PHA-induced blast transformation of T-lymphocytes was revealed. The data we obtained on the rather pronounced interferon-inducing properties of Sanogen developed by us and its combinations with well-known inductors -Cycloferon and Betaleukin, can be of practical use in the treatment of infectious pathology, especially viral hepatitis, when the combination of hepatoprotective, anti-inflammatory, antiviral, immunomodulatory and detoxification mechanisms of action, in the absence of toxicity and side effects, will ensure the development of sanogenetic processes in the patient's body.

The relationship between the character of the course of the disease, the level of tetanus and diphtheria antibodies, and indicators of general immunity in adults with infectious mononucleosis

ABSTRACT. EBV infection is one of the most common infections in the world. About 90 % of the world's population is infected with the Epstein-Barr virus. The Epstein-Barr virus is capable of affecting immunocompetent cells, causing lymphadenopathy, hepato- and splenomegaly, being stored forever in the body's lymphocytes, and causing an immunodeficient state. The aim of the study. The purpose of the study was to determine the relationships between the nature of the course of the disease for infectious mononucleosis (IM) in adults, the level of blood antibodies before tetanus and diphtheria, and general immunoreactivity. Materials and methods. In the first series of studies, blood antibodies was studied before diphtheria and tetanus in 79 adults. The control group consisted of 41 people. In the second series of studies, the immune status of 45 adults who became ill with IM was studied: 19 people who did not have a protective level of blood antibodies to diphtheria and tetanus (1st group) at the same time and 26 people (2nd group) who had a protective level of blood antibodies to these infections . In this work, the level of blood antibodies to diphtheria and tetanus toxin was studied by the ELISA method; population and subpopulation composition of blood lymphocytes using monoclonal antibodies: anti-CD3, CD4, CD8, CD22, CD16 (eBioscience, Beckman Coulter, R&D System). The proliferative activity of blood lymphocytes in the reaction of lymphocyte blast transformation was studied and the level of IgA, IgM, IgG in blood serum and the phagocytic activity of leukocytes. The results of the research were processed by the method of variation and correlation statistics using the "Statistica 10.0 for Windows" program. Results. It was established that 34.2% and 54,5 % of adults who developed MI in a moderate form did not have a protective level of blood antibodies for tetanus and diphtheria at the same time. A protective level of blood antibodies simultaneously before diphtheria and tetanus was observed in 55.2% of those who fell ill with IM in a moderate form and in 45,4 % of those who fell ill in a severe form. In persons who had a mild form of IM, the blood antibodies did not drop below the protective level before diphtheria and tetanus.The study of the immune status proved that in adults who do not have a protective level of blood antibodies against diphtheria and tetanus, the level of leukocytes, lymphocytes, CD3+, CD4+ - cells and the blast-transforming ability of T cells, the level of IgA, IgM, IgG, and the phagocytic ability of blood leukocytes are probably lower than in adults who have had an IM and have the protective blood antibodies level before these infections. Conclusions. The lack of a protective level of blood antibodies against diphtheria and tetanus in adults who have had IM is associated with a general low immunoreactivity of the body. A low level of blood antibodies before diphtheria and tetanus in persons who have had IM is closely related to the course of the disease. The lowest level of blood antibodies before these infections was observed in patients who fell ill with a severe form of IM, in patients with a mild form of the disease, significant changes in the level of blood antibodies before diphtheria and tetanus were not observed, as well as in the immune status.

Influence of Paleobacteria on the Proliferative Activity of Human Lymphocytes In Vitro

When testing the proliferative activity of 14 strains of permafrost microorganisms in the reaction of blast transformation of human lymphocytes in vitro, a strain (Alcaligenes sp.) with mitogen properties was isolated (20-fold increase in the rate of lymphocyte proliferation in comparison with the control). Four strains activated lymphocyte proliferation by 3-9 times in comparison with the control. Three strains produced substances with cytostatic properties and reduced proliferation activity by 33-43% and one strain (Bacillus sp.) almost completely suppressed phytohemagglutinin-induced lymphocyte proliferation. These data indicate that strains with a unique immunobiological potential are concentrated in the population of permafrost microorganisms that have undergone rigorous evolutionary selection.

PATHOLOGICAL IMMUNITY CHANGES IN COMORBID PATIENTS WITH EXACERBATION OF BRONCHIAL ASTHMA

The aim of the study – to investigate immunological changes in patients with exacerbation of bronchial asthma with different severity and controllability depending on the presence of comorbid pathology. Materials and methods of research. The work is based on the analysis of data from a comprehensive clinical and laboratory examination of 190 patients with exacerbation of asthma. The relative content of T–lymphocytes and their main subpopulations was determined on a flow cytofluorimeter FACS Calibur (Canada) by phenotyping lymphocytes with monoclonal antibodies (BECKMAN COULTER, USA) to surface membrane differentiation antigens (CD): CD3+19– (pan T-cells), CD4+–8+– (T−helpers/inducers), CD4–8+ (cytotoxic T lymphocytes), CD3–16+ (natural killers). To identify the imbalance of immunoregulatory subpopulations of T cells, the immunoregulatory index was calculated – the ratio of the number of CD4+8– Lf and CD4-8+ Lf. To calculate the absolute content in the peripheral blood of individual populations of Lf, leukogram indicators determined on the hematological analyzer ABH-miscros60, France were used. The proliferative activity of lymphocytes was evaluated in cultures of whole blood in the reaction of blast transformation of these cells with morphological accounting of the results. Statistical processing of the obtained data was carried out using licensed software products included in the Microsoft Office Professional 2007 package, license Russian Academic OPEN No Level № 43437596 in Excel. Conclusions. In moderate asthma, the frequency of pathological changes in the T-system of immunity did not depend on the presence of comorbid pathology, as a rule. In severe asthma, an increase in the content of pan–T cells occurred 1.2 times more often than in moderate asthma, but this was not observed in people with concomitant pathology. In this group, the frequency of increasing the content of T–helpers and decreasing the content of cytotoxic T – cells in patients with severe asthma without concomitant diseases was 1.3 times more frequent than in patients with comorbid pathology. No pathological changes in the T–system were detected in patients with controlled asthma. In partially controlled asthma, these changes were registered 1.5 times more often than in controlled asthma, and in its uncontrolled course, the prevalence of pathological changes in the T–system of immunity was much higher than in the two previous groups. In patients with concomitant diseases, these changes, as a rule, had a higher frequency than in patients without comorbid pathology.

Immune Disorders in Patients with Pulmonary Tuberculosis with Primary and Acquired Drug-Resistance of <i>Mycobacterium Tuberculosis</i>

The objective: to evaluate main parameters of cellular, humoral immunity and natural resistance in tuberculosis patients with primary and acquired multiple drug resistance; to study the correlation of these parameters with clinical and radiological manifestations.Subjects and Methods. A prospective study was conducted which included 169 patients with pulmonary tuberculosis, 80 of them were new cases of drug susceptible pulmonary tuberculosis and they made the Comparison Group (CG), and 89 patients suffered from multiple drug resistant tuberculosis and they made the Main Group (MG). The Main Group was divided into 2 subgroups: MG-1 – 40 patients with primary multiple drug resistant tuberculosis; MG-2 ‒ 49 patients who acquired drug resistance of Mycobacterium tuberculosis during treatment of tuberculosis.The immune assays included the lymphocyte blast transformation reaction (LBTR) to phytohemagglutinin and tuberculin (PPD), immunophenotyping of CD3+ and CD19+ cells; phagocytic index and phagocytic number were calculated; nitro blue tetrazolium test was used, concentrations of immunoglobulins and antimycobacterial antibodies were determined.Results. The clinical laboratory data demonstrated a more severe course of the disease in patients with multiple drug resistant tuberculosis, especially in those with acquired multiple drug resistant tuberculosis which was confirmed by the severity of immune deficiencies of the cellular immunity and innate resistance as well as by the overactivity of humoral immunity. The levels of CD3+ and LBTR reaction to PPD had a negative correlation with clinical manifestations and radiological features demonstrating the severity of pulmonary tuberculosis in patients with multiple drug resistance, especially in those with acquired multiple drug resistance; and CD19+ and antimycobacterial antibodies positively correlated with clinical manifestations and radiological signs.Conclusions. Parameters of cellular immunity and natural resistance were the lowest in the patients with acquired multiple drug resistance of Mycobacterium tuberculosis versus primary multiple drug resistance and drug susceptible Mycobacterium tuberculosis. Parameters of humoral immunity demonstrated the increased activity to compensate for the insufficiency of cellular immunity regardless of the type of resistance of Mycobacterium tuberculosis.

Functional condition of the sensitized T-lymphocytes according to RBTL in children with non-rheumatic myocarditis

Summary. The increased proportion of non-rheumatic myocarditis in children has recently determined the urgency of this problem in pediatrics. It is known that non-rheumatic myocarditis in children often occurs against the background of viral or bacterial infection under conditions of altered reactivity of the immune system. There is evidence, mainly in rheumatism, that spontaneous and stimulated lymphocyte blatransformation response in such patients reflects the activity of the process, its tendency to chronicity and may serve as a criterion for the adequacy of therapy. Studies on the functional status of T lymphocytes in patients with non-rheumatic myocarditis are mainly related to experimental models. This determines the relevance of this work. The aim was to compare the functional activity of T-lymphocytes in spontaneous and phytohemagglutenin-stimulated lymphocyte blastranformation reactions in children with non-rheumatic myocarditis in acute and chronic course. Materials and methods. Under observation were 42 children aged 4 to 13 years with non-rheumatic myocarditis. A study was made of the functional activity of T-lymphocytes in the reaction of blast transformation using a radioisotope technique with thymidine-3H. Phytohemagglutenin from Reanal was used as a nonspecific stimulant. The reaction was evaluated on an SBS-2 automatic scintillation counter. The functional activity of T-lymphocytes was studied upon admission of children to the cardiology department and after the treatment. As a result of the study of the functional state of T-lymphocytes in the blast transformation reaction using the thymidine-3H radioisotope label in 52 children with acute and chronic non-rheumatic myocarditis, it was shown that during the period of advanced clinical manifestations, high spontaneous stimulation to the nonspecific phytohemagglutenin stimulator was recorded. The complex of generally accepted therapeutic agents leads to the restoration of the functional state of T-lymphocytes in children with an acute course of non-rheumatic myocarditis, and in chronic, despite the positive clinical dynamics, increased spontaneous stimulation of T-lymphocytes is still recorded, and the response to phytohemagglutenin is not fully restored in them. These data make it possible to recommend indicators of the functional activity of T-lymphocytes as criteria for recovery and determining the timing of treatment. Conclusions. The criterion for recovery and determining the duration of treatment of children with non-rheumatic myocarditis should be not only clinical and electrophysiological indicators, but also indicators of functional activity of T-lymphocytes, reflecting the sensitization of the body

Cell-mediated immunity in patients with post-CoVID syndrome

Aim: to study the investigate of the cell-mediate immunity in post-COVID syndrome. Material and Methods: 278 patients with mild, moderate or severe Covid-19 in acute period with post-COVID syndrome were examined. All patients were questioned, and, if necessary, were examined by doctors of various specialties. The patients were divided into 2 groups depending on the severity of the course of the disease in the acute period and the duration of manifestations of post-COVID symptoms (up to 12 weeks or more). The state of cell-mediated immunity was assessed using the reaction of blast transformation of peripheral blood lymphocytes (morphological method for evaluating the reaction) in response to PHA. The activity of cells synthesizing cyclooxygenase was also determined. The control group included 200 practically healthy people of the same sex and age, who were not vaccinated and did not have a coronavirus infection. Results: a decrease in the proliferative activity of lymphocytes in response to PHA was found in patients who underwent Covid-19, both in mild (51,4±1.1%, p<0.01) and severe (49,3± 1,2% p<0,001), more pronounced in the latter in the first three months after the acute period of the disease (p<0,001). In all groups, there is an increase in the activity of cells synthesizing cyclooxygenase, which causes inflammation, which is more pronounced in patients who have had a severe form of Covid-19. Conclusion: The proliferative activity of peripheral blood lymphocytes in people who have had a coronavirus infection is reduced, both in the period up to 12 weeks and beyond; this is especially pronounced in patients who have had severe forms of Covid-19. In all groups, an increase in the activity of cells synthesizing cyclooxygenase, which plays a role in the development of inflammation, and possibly determines the progression of the course, was noted.

Comparative characteristics of the state of the immune system in patients with coronary artery disease with stable angina pectoris and acute coronary syndrome

The aim – to assess the relationship between the state of the immune system and the development of acute coronary syndrome in patients with IHD.Materials and methods. The first group consisted of 64 patients with ST-segment elevation acute coronary syndrome, mean age 54 (49–64) years; the second group – 223 patients with coronary artery disease with stable exertional angina, FC II–III, mean age 56 (49–63) years; the third group – 47 patients with acute coronary syndrome without ST segment elevation, mean age 61 (52–65) years. The material for the immunological study was peripheral venous blood. To determine the parameters of cellular and humoral innate and adaptive immunity in blood serum and supernatants of mononuclear cells, enzyme immunoassay was used.Results and discussion. In patients with coronary artery disease with acute coronary syndrome with ST segment elevation compared with patients with coronary artery disease with stable angina pectoris, the levels of indicators of the immune status in the blood were: CRP – 9.3 (5.3–12.0) versus 4.8 (2.4–8.1) mg/L (p=0.0001), sICAM – 785 (690–830) versus 565 (406–744) ng/ml (p=0.0001), IL-10 in blood mononuclear cells – 48 (1–228) versus 194 (21–758) pg/ml (p=0.0007), circulating immune complexes – 90 (70–108) versus 76 (54–105) od. (p=0.045), lymphocytes with apoptosis (CD95) – 16 (9–27) versus 11 (8–17) % (p=0.029), spontaneous oxygen-dependent metabolism of monocytes – 16 (12–21) versus 13 (9–17) (p=0.001). The levels of indicators of the immune system in the blood in patients with coronary artery disease with acute coronary syndrome with ST segment elevation compared with patients with coronary artery disease with acute coronary syndrome without ST segment elevation were: T-helpers – 37 (32–41) versus 42 (37–48) % (p=0.0006) (R=–0.33; p=0.0005), reaction of lymphocyte blast transformation to nonspecific antigen – 38 (32–47) versus 50 (42–61) % (p=0.0004) (R=–0.37; p=0.0003).Conclusions. The development of acute coronary syndrome is directly combined with increased activity of the immune system, as evidenced by the high production of proinflammatory CRP, IL-8, sICAM with a low level of anti-inflammatory IL-10, a pronounced humoral adaptive immune response (in terms of antibodies to the myocardium and vascular tissues, CD40, circulating immune complexes) and active functional state of monocytes (according to cNCT test, functional reserve, phagocytosis) in patients with coronary artery disease with acute coronary syndrome, regardless of the position of the ST segment in comparison with patients with stable coronary artery disease. Elevated levels of antibodies to the myocardium in patients with stable coronary heart disease indicate moderate myocardial damage due to temporary ischemia in angina attacks, even with a stable course of the disease. In patients with acute coronary syndrome, high levels of antibodies to the myocardium indicate myocardial damage due to increased ischemia in plaque destabilization much earlier than the clinical manifestations of acute coronary syndrome. In acute coronary syndrome with ST-segment elevation, compared with ACS patients without ST-segment elevation, activation of neutrophils and suppression of the activity of adaptive T-cell immunity is noted (by the level of T-helpers, sCD40L, blast transformation of lymphocytes, γ-interferon in mononuclear cells, apoptosis of lymphocytes).

Comparison of the effects of co-transplantation of bone marrow hematopoietic stem cells and thymic multipotent stromal cells on the immune system of mice depending on methods

Physical interaction of multipotent stromal cells (MSCs) and hematopoietic stem cells (HSCs) is a modern approach to effective and focused changes in the properties of HSCs. Resulting of those contact interaction is significant activation of cells with following immune system restoration.
 The purpose of the study is to investigate the effect of co-transplantation of bone marrow hematopoietic stem cells (HSCs) and thymic multipotent stromal cells (MSCs) separately and as a union of cells on regeneration of the murine immune system, damaged by cyclophosphamide.
 MSCs were obtained from thymuses of C57BL mice using explant technique. Bone marrow cells (BMCs) were obtained by flushing out the femur with a nutrient medium. BMCs were cocultivated for 2 hours on the monolayer of thymus-derived MSCs. The immune deficiency of mice was modelled by the treatment with cyclophosphamide (CP). After that, the cells were co-transplanted in two methods (separately into different the retroorbital sinus and as a union after co-cultivation) and the parameters of the immune system were evaluated. It was shown, that separate co-transplantation of BMCs and thymus-derived MSCs is associated with the restoration of the number of bone marrow cells, thymus, spleen and lymph nodes with an increase in the proliferation index of lymph node cells by 1.4 times compared to control. It normalized the previous reduced concentration of hemoglobin and hematocrit in the blood. Co-transplantation had a suppressive effect on the blast transformation reaction, induced by phytohemagglutinin, by 4.3 times, but showed a stimulating effect on DTHR response by 1.6 times compared to control.
 Co-transplantation of the union of BMCs and MSCs is associated with the restoration of the number of bone marrow cells, spleen and lymph nodes. The level of spontaneous apoptosis of lymph node cells significantly increased by 3.3 times compared to control. It had not effect on hematological parameters, but is activated to impact the immune system. Thus, as a result of cells union administration showed normalization of the bactericidal activity of peritoneal macrophages, unlike the separate co-transplantation. This cells graft had a suppressive effect on the number of antibody-producing cells in the spleen by 4.2 times compared to control.
 Previous co-cultivation and contact interaction of cells change the properties of cell graft. The effect of co-transplantation of BMCs and thymic MSCs is not a simple additive effect of cells. It is acquiring the features typical to certain cell types, and the expression of new characteristics. We assume this phenomenon as a result development of complex cells cooperative processes in vivo and in vitro

MICROBIAL CHARACTERISTICS OF POST-TRAUMATIC OSTEOMYELITIS

Objective: The one of the most important issues in traumatology is prevention and treatment of purulent-septic complications of traumatic diseases. The aim of our study was to establish correlations between osteomyelitis caused by bacterial flora and immunological factors. Methods: On the basis of a comprehensive study of bacteriological and immunological data in 100 patients with various etiologies osteomyelitis, using correlation analysis was determined: that the types of microbial complications following trauma and the date of the body's immune system depends on etiological factors. The frequency of microbes is different and depends on the localization of the injury and the surgical intervention. Results: frequency of the etiological factors in the contingent we studied, was distributed as follows: S. aureus-36,9%, S.Epidermidis-52,4, Ps. aeruginosa 27,4%, E. coli- 13,1%, Proteus- 27,4%. Associated infection (or co-infection e.g. S. aureus+S. Epidermidis, or St.Epidermidisis+Ps. Aeruginosa and etc) occurred in 22.6% of cases. A significantly high correlation coefficient was observed in patients who came to the clinics spontaneously or with delay, as well as with the early onset of the infectious process (up to two weeks) and surgical treatment. It also correlates positively with a decrease of following immunological parameters: NK, CD4+, CD8+ and CD 19+, the leucocytes phagocytic index is reduced and the blast transformation reaction of lymphocytes rate was increased. There is a significant correlation with benign outcome of treatment, which indicates that, the patient was sent for outpatient treatment (R=0, 79). Keywords: Trauma, Osteomyelitis, Microorganism, Etiological factor, Immune system data.

Phytochemical Profiles and In Vitro Immunomodulatory Activity of Ethanolic Extracts from Galium aparine L.

Galium aparine L., family Rubiaceae, is a widely spread species in the Galium genus. The herb of G. aparine is part of folk remedies and dietary supplements. In this study, we analyzed the chemical composition and immunomodulatory activities of G. aparine herb ethanolic extracts obtained from the plant material by maceration with 20%, 60% or 96% ethanol. The contents of hydroxycinnamic acid derivatives, flavonoids and polyphenols were determined spectrophotometrically, with extractives and polysaccharides quantified gravimetrically. The qualitative composition was studied using UHPLC-DAD-MS/MS analysis; isolation not previously described in G. aparine quercetin rhamnoglucoside was carried out through column chromatography, and the immunomodulatory activity of extracts was determined in the reaction of lymphocyte blast transformation. Major constitutes of extracts were iridoids, i.e., monotropein, 10-desacetylasperulosidic acid and asperulosidic acid; p-hydroxybenzoic acid; hydroxycinnamic acid derivatives, i.e., 3-O-caffeoylquinic, 5-O-caffeoylquinic, 3,4-O-dicaffeoylquinic, 3,5-O-dicaffeoylquinic, 4,5-O-dicaffeoylquinic acids and caffeic acid derivatives; flavonoids, i.e., rutin, quercetin 3-O-rhamnoglucoside-7-O-glucoside, and isorhamnetin 3-O-glucorhamnoside. Significantly, quercetin 3-O-rhamnoglucoside-7-O-glucoside was first isolated and identified in Galium species so far investigated. All G. aparine herb ethanolic extracts stimulate the transformational activity of immunocompetent blood cells, with 96% ethanolic extract being the most active. The data obtained necessitate further research into the mechanisms of immunomodulatory activity of extracts from G. aparine herb.

The effects of co-transplantation of bone marrow hematopoietic stem cell and thymic multipotent stromal cells on the immune system of mice during its regeneration after cyclophosphamide treatment

The multipotent stromal cells (MSCs) are considered as one of the most promising agents for regenerating the immune system due to its powerful secretion of reparative factors and immunomodulatory properties.The purpose of the study is to investigate the effect of co-transplantation of bone marrow hematopoietic stem cells (HSCs) and thymic multipotent stromal cells (MSCs) on regeneration of murine immune system damaged by cyclophosphamide.Materials and methods. MSCs were obtained from thymuses of C57BL mice using explant technique. Bone marrow cells (BMCs) were obtained by flushing out the femur with nutrient medium. The immune deficiency of mice was modelled by the treatment with cyclophosphamide. After that, the cells were co-transplanted and the parameters of the immune system was evaluated. We determined the total number of erythrocytes, hematocrit, and hemoglobin concentration in peripheral blood; phases of the cell cycle and apoptosis of the cells of the mesenteric lymph nodes; the number of antibody-producing cells in the spleen; delayed type hypersensitivity (DTH); proliferative and cytotoxic activity of natural killer lymphocytes; phagocytic activity, level of spontaneous and induced bactericidal activity of peritoneal macrophages.Results. It was shown that in contrast to BMCs, the use of MSCs alone or co-transplantation of these cells increased the spontaneous proliferative activity of lymphocytes with a significant decrease in the number of lymph node cells in G0/G1 phase by 9.2 % and an increase in the number of lymphocytes in G2-M+S phase by 35 %, as well as restoring cellularity of bone marrow, thymus and lymph nodes in mice treated with cyclophosphamide. Regeneration of erythropoiesis was stimulated by BMCs, which was manifested by the normalization of hematocrit and hemoglobin, and an increase in the number of reticulocytes in the blood by 2.2 times compared with the group of mice receiving cyclophosphamide. Co-transplantation had less pronounced, but similar effect. Transplantation of thymic MSCs stimulated the natural cytotoxicity of splenocytes by 2.7 times and substantially increased the number of antibody-producing cells in the spleen by 1.7 times compared with the group of mice receiving cyclophosphamide. Co-transplantation had a pronounced suppressive effect on the blast transformation reaction induced by phytohemagglutinin by 1.7 times, but showed a stimulating effect on DTH response by 1.46 times. Transplantation of BMCs did not affect the functional activity of the immune system.Conclusion. The effects of co-transplantation of BMCs and thymic MSCs are realized in the several parts: stimulation of hematological parameters recovery (like under the effect of BMCs separately), normalization of cell number of lymphoid organs (as under the impact of thymic MSCs); inhibition of blast transformation activity and stimulation of DTH are the effects of co-transplantation.

Lymphocytes blast-transformation reaction: modification for allergological practice

Introduction. Lymphocytes blast-transformation reaction (RBLT) is necessary for patient’s inspection with immunologic infringements. It is applied in the different fields of medicine to identification of a sensitization to antigens (allergens).Research objective – to modify and automate RBTL for application in routine laboratory practice.Materials and methods. RBTL, antigens (allergens). Software: Windows 7; Intel Pentium G4500 CPU. Digital CMOS video camera, Mikmed-6 microscope.Results. We analyzed difficulties when using RBTL in laboratory allergological practice. We created program files for automation of RBTL.Conclusion. We adapted and automated RBTL for laboratory diagnostics using.

Disorders of local pulmonary immune response in patients with chronic obstructive pulmonary diseases during exacerbation

Introduction. Morbidity due to reduced local immune reaction in patients affected by chronic obstructive pulmonary disease (COPD) is extremely high and continues to grow. The results of a research on changes in the local immune response in patients with COPD at the exacerbation stage are discussed in this paper. The pathogenesis of COPD during exacerbation is characterized by deterioration of the local innate and acquired immunity. The process is accompanied by pathological changes of the systemic immunity. Various cells of the immune response are involved in the inflammation process, attracting increased amounts of macrophages, neutrophils, T- and B-lymphocytes, and dendritic cells. The number of neutrophils and B-lymphocytes are observed in patients suffering from a severe disorder. Most common symptoms of COPD are irreversible narrowing of the airways, destruction of the parenchyma of the lung accompanied by emphysema, and progression of the obstructive pulmonary disease. The aim of the study – to learn the changes in functionality of bronchoalveolar lavage (BAL) cells and peripheral blood cells (PBC) in COPD patients and provide an explanation for these changes in light of pathogenetic characteristics to predict the course of the disease and justify the necessity of a local immunotherapy in comprehensive treatment of COPD patients. Research Methods. The focus of the research was on BAL cells and PBC of patients with COPD at different stages and on blood serum. CD3, CD4, CD8, CD16 and CD19 of BAL and PBC cells were identified using different specific monoclonal antibodies. Receptor expression to IL-2 (CD25) was examined, adhesive molecules were identified using CD54, proliferation levels were measured using IPO-38, as well as was measured the expression of class II antigens MHC (HLA-DR) in the blast transformation reaction that was induced by PHA and Con A mitogens. Phagocytic activity of phagocytizing neutrophils cells, alveolar macrophages (AM), and phagocytizing PBC cells was evaluated. Levels of free radical processes and antioxidant defence were studied using a chemiluminescence method. Results and Discussion. The amount of AM cells in COPD patients declined significantly, whereas the amount of neutrophils notably increased, as well as concentrations of the myeloperoxidase and the eosinophil cationic protein. Examination of phlegm and BAL cells obtained from COPD patients at the exacerbation stage of the disease indicate that 23 % of samples displayed gram-negative bacterial flora (primary pathogens being Pseudomones aeruginosa, Haemophillus influensae, Enterobacter spp.), 15 % – candidas, 61 % – pathogen type was not identified. Researching local and systemic immune response in COPD patients and functional activity changes of BAL and PBC indicated that patients showed significant changes in their local immune system, which manifested in significant reduction in the amount of neutrophils in the lavage, reduction in CD3, CD19, and increased expression of receptors to HLA-DR antigen expression. Summary. Parallel study of the local and systemic immunological responses contributes to expand our knowledge and understanding of pathogenesis of COPD. It is useful in determining methods to help decrease the number of complications and aid in COPD prophylaxis in persons of working age. It also provides information on which local immunotherapy will prove to be most effective. Exploring the subject further should provide fundamental understanding of similar and different symptoms of various disorders in patients with inflammation processes at the local immune level in different body locations, such as respiratory organs during COPD, in the peritoneal cavity during peritonitis, dermal lesions or any other changes in the local immune system of a different body location.

Characterisation and In Vivo Safety of Canine Adipose-Derived Stem Cells

Abstract The study characterises canine adipose-derived stem cells (cASCs) in comparison to human ASCs (hASCs) and tests their safety in a canine model after intravenous administration. cASCs from two dogs were cultured under hypoxic conditions in a medium supplemented with autologous serum. They were plastic adherent, spindle-shaped cells that expressed CD73, CD90, and CD44 but lacked CD45, CD14, HLA-DR, and CD34. cASCs differentiated toward adipogenic, osteogenic, and chondrogenic lineages, although adipogenic differentiation capacity was low. Blast transformation reaction demonstrated that these cells significantly suppress T-cell proliferation, and this ability is dose-dependent. Intravenous administration of a cell freezing medium, therapeutic dose of cASCs (2 × 106 live cells/kg), and five times higher dose of cASCs showed no significant side effects in two dogs. Microscopic tissue lesions were limited to only mild, non-specific changes. There were no signs of malignancy. The results of the study indicate that cASCs are similar to hASCs and are safe for therapeutic applications in a canine model. The proposed methodology for ASC preparation on a non-routine basis, which includes individually optimised cell culture conditions and offers risk-adapted treatment, could be used for future personalised off-the-shelf therapies, for example, in myocardial infarction or stroke.

Реакція бластної трансформації лімфоцитів крові нелінійних самиць щурів, їх приплоду на ранніх етапах постембріонального розвитку на фоні впливу сольового екстракту Hirudoverbena

Реакція бластної трансформації лімфоцитів крові нелінійних самиць щурів, їх приплоду на ранніх етапах постембріонального розвитку на фоні впливу сольового екстракту Hirudoverbena

Lithium Salts of Krebs Cycle Substrates as Potential Normothymic Antioxidant Agents.

Objective:Aim of the present work was to study the antioxidant properties of lithium salts of Krebs cycle substrates and their influence on immune cells. Lithium is a well-known and widely used mood stabilizer. These lithium-based substances have a lot of potential properties because of the anionic component of the Krebs cycle substrates, which take part in basic intracellular biochemical process.Materials and Methods:Lithium salts of fumarate, pyruvate, malate, succinate, and citrate (as reference drug) were investigated in this study as antioxidants and immunomodulators. The antioxidant properties were studied by the voltammetry method, which evaluates oxygen radical scavenging capacity of lithium substances. Influence of the lithium compounds on the immune cells of human blood was indicated by the reaction of blast transformation of lymphocytes.Results:All tested substances and their mixes possessed antioxidant properties, more expressed in maximal therapeutic concentration. Lithium compounds showed no toxic influence on human blood immune cells and caused no significant changes in both spontaneous and stimulated proliferation.Conclusion:The results allow considering lithium salts of Krebs cycle substrates as potential normothymic agents (mood stabilizer) with antioxidant properties and low toxicity.