Functional Activity Of Immunocompetent Cells Research Articles

Vitamin D supply, respiratory viral infections and persistent wheezing in children: how significant is the relationship?

The review discusses the multifaceted effect of vitamin D on antiviral immunity, induction of antimicrobial peptides, functional activity of immunocompetent cells, autophagy processes, clearance of viruses and viral fragments. The relationship between vitamin D and allergic inflammatory cells, its modulating and tolerogenic potential is considered. The inhibitory effect of vitamin D on eosinophils, mast cells, the production of pro-inflammatory and proallergic mediators, and the synthesis of IgE has been shown. The protective role of vitamin D in the development of remodeling and hyperresponsiveness of the respiratory tract has been established by reducing the number of goblet cells, mucus hypersecretion, and the expression of metalloproteinases. The effects of vitamin D on the replication of respiratory viruses and experimental resistance to them through the induction of antimicrobial peptides and other mechanisms have been demonstrated. Data from clinical studies on the preventive effect of vitamin D on the incidence of respiratory infections and episodes of persistent wheezing in children are presented. Children with respiratory syncytial infection and vitamin D deficiency were more likely to require respiratory support and mechanical ventilation. The preventive role of vitamin D in the development of obstructive bronchitis in children and severe bronchiolitis in infants has been shown. It has been established that normal levels of 25(OH)D in the mother significantly reduce the risk of developing bronchial obstruction in the child. Alternative data that do not support a protective effect of vitamin D on respiratory morbidity and wheezing in children are discussed. Further monitoring studies that take into account geographic location and analysis of the dose of cholecalciferol used to assess its preventive role in the development of wheezing and bronchial asthma in children are advisable.

Estimation of the effect of lithium salts on cytokine production by blood cells in in vitro experiments

Aim. To study the effects of lithium salts on production of cytokines by immunocompetent cells in the whole-blood culture of patients with alcohol dependence and affective disorders.Materials and methods. The study materials were blood samples from 25 patients with alcohol dependence (AD) and 12 patients with bipolar disorder (BD). Blood diluted 1:1 with complete RPMI-1640 medium (Gibco, UK) was added to the wells of the culture plate, then new lithium salts (succinate, fumarate, pyruvate, ascorbate) and a reference salt – lithium carbonate at a final concentration of 1.2 mmol / l per lithium ion – were added. In parallel, control samples without lithium salts were tested; the samples were incubated for a day. The concentration of cytokines (interferon (IFN) γ, interleukin (IL)-1β, IL-2, IL-4, IL-6, IL-8, IL-10, IL-17A, tumor necrosis factor (TNF) α) was determined in the culture supernatants on the MAGPIX multiplex analyzer (Luminex, USA) (Center for Collective Use “Medical Genomics”, Tomsk NRMC) using the Human Cytokine / Chemokine Magnetic Bead Panel (Merck, Germany).Results. All lithium salts had a unidirectional effect on the production of cytokines by immunocompetent cells (ICC), except for lithium ascorbate and IL-8. The concentrations of cytokines in the supernatants of loaded and control samples (spontaneous production) were comparable, which indicates an absence of stimulating or suppressing effects of salts on the functional activity of ICC under the experimental conditions. The effect of lithium ascorbate as an IL-8 inducer was detected: the production of IL-8 induced by lithium ascorbate was 2.3–2.5 times higher than its spontaneous production.Conclusion. The obtained results, as well as the previously revealed antioxidant and cytoprotective properties of new lithium salts, confirmed that they are promising for development of pharmacological agents with combined action.

Number of T- and B-lymphocytes and their functional activity in the blood of common carps affected by Aeromonosis, and in the treatment of probiotic Enteronormin

Fish bacterial infectious diseases are a significant problem for aquaculture, often leading to massive fish deaths and significant economic losses. For fish treatment often used antibacterial drugs: nitrofuran, antibiotics, feed antibiotics, as well as dyes. Finding effective drugs and studying their effects on the body of fish are relevant. Nowadays, different groups of substances, including probiotics, are offered as an alternative to increase the growth of animals and to combat pathogens. Here are the results of studies of the influence of the probiotic drug "Enteronormin" on the indicators that characterize the status of T- and B-cell immunity of carp affected by bacterial Aeromonosis. Enteronormin is a probiotic supplement , wich contains a bacteria complex - Enterococcus spp, Bacillus subtilis spp, Lactobacillus spp; and excipients - chitosan, peptone enzymatic and ground sugar (RP № BB-00427-02-12 dated 13.04.2012). The research was conducted under the conditions of aquariums at the Lviv Research Station of the Institute of Fisheries of NAAS. The control group, which consisted of clinically healthy fish, received only 3% starch suspension, the first experimental group (D1) - Aeromonosis affected carps who were given only 3% starch suspension, the second experimental group (D2), which consisted of carps affected Aeromonosis, through the probe within 7 days was administered the drug "Enteronormin", at the rate of 2 mg per 1 kg of fish weight, in the composition of 3% starch suspension. Before feeding for the carp drug was activated for 14-16 hours water enriched with Iodine and Selenium ions in the Jodis + Se form (TU U 15.7-30631018-011: 2011). It is established that the disease of carp on Aeromonosis leads to changes in the number and functional activity of immunocompetent blood cells. The inhibition of lymphocytogenesis and functional activity of immunocompetent cells in carp blood in a disease associated with the bacterial form of aeromonosis was ascertained. The use in the experimental carps group in the composition of 3% starch suspension of the drug "Enteronormin" had a normalizing effect on the state of T-cell specific protection, but did not significantly affect the amount of EAC-RUL and their functional activity. Key words: fish, carp, aeromonosis, probiotics, Enteronormin, T-lymphocytes, B-lymphocytes.

Оценка эффективности противовирусной терапии гриппа A(H1N1) в эпидемические сезоны 2017 – 2018 и 2018 – 2019 гг.

Представлены результаты оценки терапевтической эффективности риамиловира в сравнении с умифеновиром по длительности общих инфекционных синдромов (интоксикации, лихорадки), синдромов поражения респираторного тракта, а также по частоте развития осложненного течения гриппа. Для оценки функциональной активности иммунокомпетентных клеток на разных этапах заболевания при проведении противовирусной терапии проводилась оценка показателя лимфоцитарно-тромбоцитарной адгезии (ЛТА). Установлено, что у пациентов, получавших риамиловир, количество розеткообразующих лимфоцитов снижалось до (16,8 ± 2,1) %, что в 1,2 раза меньше по сравнению с группой пациентов, принимавших умифеновир (21,2 ± 1,8) %. В результате применения риамиловира при гриппе происходил не только более быстрый регресс интоксикационного и катарального синдромов (p < 0,05), но и нормализация ЛТА, что отражает стабилизацию патологического процесса и, соответственно, свидетельствует о большей эффективности проводимой терапии данным противовирусным препаратом.

Вплив вітамінів Е і С на кількість і функціональну активність τ- ι β-лімфоцитів крові курчат-бройлерів

Abstract . Separate indices of T- and B-cell immunity of broiler chickens were investigated during the period of their growth with additional introduction of tocopherol acetate and ascorbic acid into their diet. The studies were conducted in one of the farms in the Lviv region on four groups of 100 broiler chickens each, ranging from 1- to 41 days of age. Chickens in the control group received a standard diet, the first experimental diet with the addition of tocopherol acetate, the second experimental - ascorbic acid, and the third - a complex of these vitamins. In whole blood, the relative number of T and B lymphocytes and their individual populations were determined in the reaction of spontaneous rosette formation with ram erythrocytes, differentiated calculation of rosette lymphocytes with different degree of functional activity was performed. For research, blood was collected from broiler chickens after slaughter at different ages: 27-, 34-, and 41-day-olds. During the work, the bioethical requirements of the animals were complied with according to the current legislation. Studies have shown that the additional introduction of chickens to their diet tocopherol acetate and ascorbic acid has a stimulating effect on the number of T and B lymphocytes and the functional activity of T and B cell units of immunity. This is evidenced by the higher number of (-lymphocytes (common, active, theophylline-resistant) and Β-lymphocytes in the blood of broiler chickens, which were supplemented with vitamins E and C. Τ- and Β-lymphocyte apparatus - an increase in the number of cells with low and medium degrees of aperture and a decrease in the inactive function of Τ- and Β-lymphocytes in the blood. These changes were more pronounced in the blood of chickens at 27 days of age due to the action of vitamin C, as well as the complex of tocopherol acetate and ascorbic acid. Therefore, on the basis of the conducted researches it is possible to confirm a regulatory role of vitamins E and C in formation of a cellular link of specific immune response of chickens-broilers. The action of the investigated vitamins, and especially of ascorbic acid, revealed an increase in the number and increase in the functional activity of T- and B-lymphocytes of the blood. These changes are probably due to the antioxidant action of vitamins in providing metabolic homeostasis of membranes and the receptor apparatus of immunocompetent cells. Key words: vitamins E and C, broiler chickens, specific immunity, T and B lymphocytes, functional activity of immunocompetent cells

ЭКСПЕРИМЕНТАЛЬНОЕ ИЗУЧЕНИЕ ИММУНОТРОПНОЙ АКТИВНОСТИ АЛБЕНДАЗОЛА В НИЗКИХ ДОЗАХ

Evaluation of the immunotropic effect of albendazole in low therapeutic doses has been carried out based on testing the ability of CBA × C57BL/6 mice lines to induce a delayed-type hypersensitivity (DTH) and the synthesis of antibodies against the thymus-dependent antigen. It has been shown that albendazole at doses of 2.5 and 5.0 mg/kg had no effect on the immune response of the cellular and humoral type. The DTH indices were 10.1 ± 3.14 and 10.8 ± 2.45%, respectively, and slightly exceeded those in the control by 3.1 and 10.2%. When albendazole was administered to immunized animals at doses of 2.5 and 5.0 mg/kg, log1/2 of the blood hemagglutinin titer was 6.8 ± 0.46 and 7.0 ± 0.47, and effect indices of albendazole were 1.03 and 1.06. The titer of hemagglutinins in the blood of experimental animals exceeded that in the control by 3.0 and 6.1%, respectively. It has been shown that immunocompetent cells, which functionally provide the formation of delayed-type hypersensitivity and the synthesis of antibodies against the thymus-dependent antigen, are tolerant to albendazole. Albendazole at low therapeutic doses of 2.5 and 5.0 mg/kg with a single administration did not have immunotropic activity. At the tested doses, albendazole is not a regulator of cellular and humoral processes since it does not contribute to modulation of the functional activity of immunocompetent cells and their cooperative interaction, which are necessary for the development of an immune response of the cellular and humoral type.

Immunobiological aspects of cow lactation

Lactation of cows is a high-energy metabolic process. It is regulated in the body by the complex physiological mechanisms of the nervous, endocrine and immune systems. The purpose of to study changes in the immunobiological reactivity of the body of cows during lactation. Clinical and experimental studies were accomplished at the veterinary clinic and in the specialized laboratory of reproductive animal immunology. For the study, four groups of analogues of experimental animals were formed in which the immunological methods determined the immune status: cows (n = 17) during the early lactation (secretion of colostrum, 3–5th day); second (n = 32) – cows (n = 32) in the mid (3–5th month) lactation; the third (n = 28) – during the late lactation (5–7th day) and the fourth group (n = 28) – during the dry period (12–20th day). Our research has shown that during lactation in the body of cows fluctuations of the studied indicators of cellular and humoral immunity occur. Experimental data that showed that the percentage of T-lymphocytes in the blood of cows of Ukrainian dairy black-billed breed during the initial (3–5 days) period of lactogenesis is the lowest (37.88 ± 1.53%). However, the lowest expression of B-immune cells (16.05 ± 0.74%) was also detected during colostrum secretion. These changes occurred against the background of the decrease in CD3+/СD22+ (2.36 ± 0.11). In particular, at the 3–5th month of lactation, peripheral blood flow contained 53.40 ± 0.83% CD3+, ie 1.40 times more. The number of CD22+ mononuclear cells increased by only 1.14 times (up to 18.31 ± 0.69%). The beginning late lactation and dry period in the number of immunocompetent cells was observed: CD3+ – up to 42.11 ± 1.03%, and CD22+ – up to 22.92 ± 0.89%. All this was accompanied by a corresponding redistribution of the balance (CD3+/СD22+). We have identified certain patterns in the blast transformation ability of T-lymphocytes, which was manifested in the change in the activity of activated immune cells of blood. At the beginning of lactation, the functional capacity of T cells was the lowest (42.47 ± 0.62%). But subsequently with the increase of the lactation curve there was a certain activation of their transformation into blasts (51.11 ± 1.05%). At the beginning of late lactation cows underwent a gradual decrease in the blasttransformation capacity of immunocompetent cells (43.11 ± 0.89%). And which continued to decline gradually during the dry period. Thus, during lactation in the body of cows there are dynamic permanent changes in the system of cellular and humoral immunity. From the beginning of lactogenesis, there is a gradual increase in the increase in the functioning of the mammary gland of cows, and further decrease in the population composition of CD3+ and СD22+, which was also accompanied by changes in the functional activity of immunocompetent cells.

Correction of immune disorders and changes in the protein/lipid spectrum of circulating erythrocyte membranes in adenomiosis

Hypothesis/aims of study. Disorders of immune homeostasis, morphofunctional and ultrastructural changes of erythrocytes in endometriosis lead to hypoxia, impaired micro-rheological and hemostatic statuses, dysmetabolism, organ and system dysfunction, which may contribute to the systemic consequences of this disease. The aim of this study was to evaluate the immune status and protein/lipid spectrum of erythrocyte membranes, as well as to develop methods for the pharmacological correction of impaired parameters in patients with internal endometriosis.
 Study design, materials, and methods. Fifty-seven patients with verified grade I–II adenomyosis were examined, assessing some immune status parameters and determining protein and lipid fractions of circulating erythrocyte membranes before and after pharmacotherapy.
 Results. The changes in the immune status parameters and protein and lipid composition of erythrocyte membranes revealed in adenomiosis disrupt the functional activity of immunocompetent cells and erythrocytes.
 Conclusion. The proposed methods of pharmacotherapy are pathogenetically substantiated and contribute to the correction of both the chosen immune status parameters and structural and functional properties of red blood cells, while improving the quality of treatment of patients with adenomyosis.

Monoclonal antibodies of IKO series in the three-color analysis of sub-populations of peripheral blood T-lymphocytes of oncological patients

Background.Clinical studies have shown that the most oncological patients have impaired immunological reactivity. Monitoring of the immune system status of patients with solid tumors includes a subpopulation analysis of immunocompetent cells. It is based on the study of expression of surface antigens and intracellular structures, and an analysis of the functional activity of immunocompetent cells. Today two, three, and even four-color reagents are used for subpopulation analysis of lymphocytes. Fluorescent conjugate with monoclonal antibodies (MAB) of the ICO series and fluorescent dyes Imd-306, Imd-506 (cyanine) and phycoerythrin (a group of phycobiliproteins) was received and characterized in N.N. Blokhin National Medical Research Center of Oncology. These conjugates have proved themselves in one and two-color analysis of subpopulations of peripheral blood lymphocytes by flow cytometry.The purposeof this study is to assess the possibility of sharing several immunofluorescent probes (IFP) on the basis of different Mab series of ICO and dyes FITC, Imd306, Imd506 and PE for three-color analysis of lymphocyte subpopulations of donors and patients with oncological diseases by flow cytometry method.Materials and methods.A panel of immunofluorescent probes (IFP) CD3FITC, CD4PE, CD8-Imd506, CD4-Imd306 was used in the work. Probes are obtained with MAB (ICO series) and fluorophores FITC, PE, Imd-306, Imd-506. The specific activity of the probes was evaluated in the immunofluorescence reaction by flow cytometry. Donor’s peripheral blood was used to optimize the conditions of the immunofluorescence reaction. Clinical approbation of the obtained sets was carried out on 2 groups of patients with oncological diseases before and after surgical treatment. The first group included 64 patients with cancer of the oral mucosa. The second group consists of 35 patients with ovarian cancer.Results. We have shown that for the three-color analysis of human lymphocytes by flow cytometry, it is recommended to use sets of labeled MAB: CD3-FITC (10 μg/ml) + CD4-Imd306 at a concentration of 5–10 μg/ml; CD3-FITC + CD4-PE at a concentration of 20 μg/ml and CD3-FITC + CD8 Imd506 at a concentration of 10 μg/ml. The final concentration of immunoglobulin in the solution of IPS is indicated. It was shown that even before the beginning of treatment in patients with cancer of the oral mucosa the subpopulation CD3+ CD4+ content was significantly lowered in the structure of T cells, and the total level of CD8+ lymphocytes significantly exceeds the parameters of the donor group. Analysis of linear CD markers of lymphoid cells in patients with ovarian cancer revealed a significant decrease in the total number of CD3+ T-lymphocytes in comparison with donors. There was no evidence of the effect of surgical treatment on the subpopulation structure of lymphoid cells.Conclusion.Combinations of fluorescent probes CD3-FITC/CD4-Imd306/CD8-Imd506 and CD3-FITC/CD4-PE/CD8-Imd506 can be used for trichromatic analysis by flow cytometry of the lymphocyte subpopulations of both healthy donors and cancer patients. Comparative studies of the specific activity of combinations of IFP based on Mab ICO series and reference commercial test systems (BD Biosciences) on lymphocytes of healthy donors and oncological patients (ovarian cancer, oral mucosa cancer) showed comparable results. The application of the obtained probes made it possible to reveal violations of the subpopulation structure of peripheral blood lymphocytes in patients with ovarian cancer and cancer of the oral mucosa, in particular, CD3 – /CD8+, CD3+/CD4+, CD3+.

IN VITRO INDUCTION OF BLOOD MONONUCLEAR CELL PROLIFERATION BY ENDOGENOUS RETROVIRAL HERV-Eλ4-1 ENVELOPE PEPTIDE IN PATIENTS WITH MULTIPLE SCLEROSIS

A comparative in vitro study of blood mononuclear cells from multiple sclerosis patients and healthy donors was performed, in order to evaluate proliferative response to a retroviral antigen, aiming to determine immunomodulatory properties of synthetic oligopeptide homologous to a highly conserved human endogenous retrovirus HERV-Eλ4-1 envelope protein. It was revealed that this oligopeptide is able to stimulate the in vitro spontaneous and mitogen-induced proliferation of blood mononuclear cells from either donor and multiple sclerosis patients. Intensity of this oligopeptide-induced stimulatory effect depends on the protein concentration, and on initial level of blood immunocompetent cells proliferation. Hence, the endogenous retrovirus HERV-Eλ4-1 envelope region protein is able to increase functional activity of immunocompetent cells from human blood, that suggesting its immunostimulatory properties. It is possible that the mitogenic effects of this protein upon immunocompetent cells of multiple sclerosis patients represent a potential mechanism of retroviral involvement into pathogenesis of the disorder.

Influence of Tularemia Microbe of Different Subspecies on the Functional Activity of Immunocompetent Cells of Experimental Animals

Influence of Tularemia Microbe of Different Subspecies on the Functional Activity of Immunocompetent Cells of Experimental Animals

Monitoring immunosuppression with measures of NFAT decreases cancer incidence

Long-term immunosuppression causes a significantly increased risk for the development of malignancies in transplanted patients. A link between immunosuppression and incidence of cancer is well documented and involves the effect of immunosuppression on anti-tumor surveillance and antiviral adaptive immune responses. We present a 67-year-old patient with a history of recurrent non-melanoma skin cancer. After adjustment of immunosuppressive therapy under close pharmacodynamic control, the development of new malignant lesions could be prevented. The availability of a quantitative, quick laboratory test for an assessment of the individual functional activity of immunocompetent cells that are crucial for transplant rejection, defense against viral infection, and tumor surveillance along with the ability to adjust doses of immunosuppressive agents such that patients are largely protected against malignant disease and/or viral infection are important. NFAT-regulated gene expression measured in peripheral blood allowed us to predict "safe" immunosuppression. Thus patients could maintain a stable allograft function. This represents a breakthrough in transplantation medicine and advances our attempts to individualize treatment in transplanted patients.

Effects of recombinant IL-4δ2 on human peripheral blood mononuclears

In human cells, expression of IL-4 gene involves alternative mRNA splicing. IL-4delta2 splice variant is an antagonist of full-length IL-4 protein and blocks its effect on the functional activity of immunocompetent cells. The effect of recombinant IL-4delta2 on cytokine-producing activity of human peripheral blood mononuclear cells is shown for the first time.

Immunity Parameters in Mice of Different Strains

Quantitative composition and functional activity of immunocompetent cells differ in mice of different strains. The counts of T cells in the bone marrow and spleen, proliferative activity of T cells in the spleen, levels of IL-2 and IL-10 production by splenic T cells, number of antigen-specific T cells and their functional activity are low in C57Bl/6, BALB/c, and CC57W mice and high in CBA/CaLac, DBA/2, and C3H animals. Low phagocytic activity of peritoneal macrophages was detected in BALB/c and CC57W mice and high activity in C3H animals. The content of antibody-producing cells in the spleens of C57Bl/6, BALB/c, and CC57W mice is higher than in CBA/CaLac, DBA/2, C3H, A/SN, and AKR/JY mice. Functional activity of B cells is lower in BALB/c and CC57W compared to CBA/CaLac and DBA/2 mice.

Saquinavir Up-Regulates Telomerase Activity in Lymphocytes Activated with Monoclonal Antibodies Against CD3/CD28

The present study describes the effect of the HIV protease inhibitor saquinavir on telomerase activity and interferon-γ (IFN-γ) production of non-adherent mononuclear cells (NA-MNC). Cells obtained from peripheral blood of healthy donors were exposed in vitro to a mixture of monoclonal antibodies against CD3 and CD28 membrane antigens in order to activate prevalently T cell subsets. Treatment with saquinavir was performed at the time of cell stimulation. Thereafter, NA-MNC were tested for telomerase activity (TRAP assay) and interferon-γ production up to 7 days later. The results show that saquinavir up-regulates telomerase activity and IFN-γ release in activated NA-MNC. These observations suggest that the anti-HIV effects of saquinavir could be accompanied by other immunopharmacological properties, influencing some aspects of the functional activity of immunocompetent cells. These include possible antagonistic effects against lymphocyte senescence, through telomerase activation, and a potentiating activity on the production of IFN-γ following T cell activation.

Functional activity of immunocompetent cells in patients with anthracosilicosis

The most important link in the pathogenesis of pneumoconiosis, considered from the standpoint of immunopathological theory, is the unconditional and decisive decay of macrophages absorbing silica particles, which induce a response at the level of immunocompetent cells: T- and B-lymphocytes (A. E. Vershigora, 1980).