Cytokine production by peripheral blood lymphocytes in patients with ovarian epithelial tumors

In presence of autoserum, production of IL1b, IL2, IL4, IL6 and TNFa by peripheral blood lymphocytes of 30 patients with epithelial tumors of ovaries and 50 patients with epithelial ovarian cancer of I-IV stages was investigated. The data received enable to ascertain the increase of mononuclear activity during ovarian tumor progression, increase of T-helpers of the second type activity, along with the reduction of T-helpers of the first type activity in peripheral blood of patients, and abnormalities of IL6-dependent mechanism of control of ILIb and TNFa production by peripheral blood lymphocytes of patients with ovarian cancer.

Changes of immune status in patients with different PD-L1 expression after stereotactic body radiation therapy of oligometastasis

Purpose. To study the influence of PD-L1 expression on the dynamics of immunological changes before and at different intervals after stereotactic body radiation therapy (SBRT) of metastatic lesions in patients with metastatic forms of solid tumors. Materials and methods. A quantitative assessment and analysis of blood immunological parameters was conducted before irradiation, via 3-4 weeks and via 6-8 weeks after SBRT in patients with malignant tumors with oligometastases in the liver or lungs, in groups with negative and positive expression of PD-L1. All peripheral blood samples were analyzed by flow cytometry. Statistical analysis was performed using Friedman and Nemenyi criteria. Results. 3-4 weeks after the end of SBRT in group CPS <1 we observed statistically significant increase of activated T-helpers (CD3+CD4+HLA-DR+), activated cytotoxic T-lymphocytes (CD3+СD8+HLA-DR+), T-lymphocytes (CD3+CD19-) and T-helpers (CD3+CD4+). Wherein, activated T-helpers and activated cytotoxic T-lymphocytes statistically significantly increased 6-8 weeks after SBRT compared with the study before irradiation. In group CPS> 1, we revealed statistically significant increase of activated T-helpers 6-8 weeks after and decrease of T-regulatory lymphocytes (CD4+CD25brightCD127low) 3-4 weeks after completion of SBRT compared with the study before radiotherapy. When we analyzed the indicators by the TPS index, most of the statistically significant changes were recorded in the group with negative expression (TPS <1): increasing of activated T-helpers and activated cytotoxic T-lymphocytes 3-4 weeks and 6-8 weeks after SBRT and decreasing of T-regulatory lymphocytes 3-4 weeks after irradiation compared with the study before irradiation. Conclusion. Groups with negative PD-L1 expression (CPS <1 and TPS <1) are associated with a more activated antitumor T-cell immune response compared to patients with positive PD-L1 status (CPS≥1 and TPS≥1), however, further researches are needed.

Comparative characteristics of different methods of treatment of microsporia

Microsporia is the most common disease of fungal etiology, most often caused by the pathogen Microsporum canis. Treatment of this disease requires a comprehensive approach, because the disease is dangerous to humans. Therefore, in the treatment of microspores, it is important not only to carry out therapeutic measures, but also to prevent the spread of the pathogen of the fungus in the environment and increase the immune status of the organism in the fight against infection. The main source of infection is cats. In order to determine the effectiveness of treatment of microsporia by various methods, studied of the blood and skin of guinea pigs infected with the pathogen M. canis. Sick animals were divided into three groups. The first group was treated with the systemic antifungal itraconazole and topical treatment with a solution of clotrimazole. Treatment of the second group was performed with a topical antifungal agent (1% solution of clotrimazole) with vaccination with the antifungal vaccine “Vakderm”. For the third group, the developed drugs were used – antifungal agent “Micromar” and immunostimulant “Biogluk”. During treatment, hematological and immunological blood researches and histological skin examinations were performed. During treatment with antifungal drugs (itraconazole and clotrimazole) the number of leukocytes decreases from 11.13 ± 0.72 to 7.13 ± 0.22, rod-shaped neutrophils from 15.76 ± 1.29 to 5.50 ± 0.76, and segmental increases from 12.17 ± 1.47 to 24.17 ± 2.27, decreases ESR from 5.67 ± 0.67 to 2.33 ± 0.42, which occurs when inhibiting the inflammatory response of the organism to infection. Thrombocytopenia (from 231.17 ± 7.60 to 184.33 ± 7.65) and eosinophilia (from 2.70 ± 0.73 to 7.33 ± 1.33) are also noted. There is a slight increase in T-helpers and a decrease in T-suppressors. Histologically, the infiltration of the dermis by histiocytes and eosinophils under skin persists. In the treatment of microsporia by treatment with 1 % solution of clotrimazole and vaccination with the vaccine “Vakderm” the results of studies showed that the number of leukocytes decreases from 11.13 ± 0.72 to 5.35 ± 0.31 (P < 0.01), rod-shaped neutrophils from 15.76 ± 1.29 to 7.67 ± 0.56, and segmental increases from 12.17 ± 1.47 to 22.17 ± 0.91 (P < 0.001), decreases). The number of T-helpers is increasing. The histological picture on day 7 is characterized by hyperkeratosis, and on day 14 the hyperemia of the basal layer of the epidermis persists. When using the antifungal drug “Micromar” and immunostimulant “Biogluk” the results of studies showed that the number of leukocytes decreases from 11.13 ± 0.72 to 6.95 ± 0.10, rod-shaped neutrophils from 15.76 ± 1.29 to 6.17 ± 0.65, and segmental increases from 12.17 ± 1.47 to 22.00 ± 0.86, decreases ESR from 5.67 ± 0.67 to 2.17 ± 0.31. Increases the number of T-helpers and the number of natural killers and T-suppressors is gradually decreasing. Histological changes are presented in the form of dilation of blood vessels and visualization of single erythrocytes in the dermis on day 14 of treatment.

Immune status of young pigs different methods of their breeding using means Globigen® Pig Doser and Globigen® Jump Start

The article deals with the influence of the use of means for young pigs Globigen® Pig Doser та Globigen® Jump Start, which due to the content of specific Ig Y have immunostimulatory properties. Assessing the level of nonspecific resistance of piglets, which are based on determining the content of T- and B-lymphocytes and their populations in the blood of animals 30-, 60- and 120-day-old, the trend of increasing the number of T-helpers, T-natural killers and B-lymphocytes in the experimental groups of animals of both farms. It was found that the content of T-helpers during single-phase rearing of pigs became more important under the action of Globigen® Pig Doser and ranged from 30.16−35.15 %, and during three-phase rearing – under the action of Globigen® Jump Start 34.58−35.08 %. An increase in the content of B-lymphocytes to the level of physiological norm was noted: FE PE “Glynjany Agro” (single-phase cultivation) in the first experimental group 16.75–19.45 %, in the second 17.66–21.11 %. At Ltd. “Meat Resources” (three-phase cultivation) 17.7–21.02 % and 18.91–21.01 %, respectively, in the first and second experimental groups. Regarding the indicators of the content of T-natural killers, there is a tendency to increase their number in the blood of animals of experimental groups by an average of 4.90 % for single-phase rearing and 3.09 % for three-phase rearing of pigs. Such data give grounds to claim that there is a formation of the immune response to the action of pathogens, which plays an important role in maintaining the barrier function of the intestinal mucosa, which in turn reduces mortality and increases the average daily gain of young pigs. Under the action of immunostimulants, the safety of piglets in FE PE “Glynjany Agro” (single-phase rearing) at 30 days of age increased to 81.3−93.7 %, and in Ltd. “Meat Resources” (three-phase rearing) to 80.0−86.7 %. In piglets FE PE “Glynjany Agro”, up to 120 days of age, the average daily gain was 0.710 and 0.691 kg using Globigen® Pig Doser and Globigen® Jump Start, and in piglets from Ltd. “Meat Resources” these figures were 0.595 and 0.628 kg.

T-lymphocytes phenotypic composition of peripheral blood in patients with Graves’ disease undergoing conservative therapy with thiamazole

BACKGROUND: Effective control of autoimmune inflammation in Graves’ disease determines necessity to study the T helper (Th) and cytotoxic T-lymphocytes dysfunction, as well as the level of regulatory T-cells (Treg) activation in patients with Graves’ disease on thyrostatic medication, which will clarify the immunomodulatory effects of long-term thiamazole treatment serve as targets for more specific therapies.AIM: To study the phenotypic composition of T-lymphocytes in the peripheral blood of patients with Graves’ disease to assess the direction of immune response depending on thimazole-induced euthyroidism duration.MATERIALS AND METHODS: A single-center, cohort, continuous, open-label, controlled trial was conducted to assess the phenotypic composition of T-lymphocytes in peripheral blood in women with Graves’ disease on long-term thiamazole treatment. The phenotypic composition of T-lymphocytes was determined by flow cytometry using direct immunofluorescence with conjugated FITC monoclonal antibodies depending on the duration of thimazole-induced euthyroidism of long-term thiamazole treatment.RESULTS: The study included 135 women with Graves’ disease, mean age 43.09±12.81 years, 120 (88.91%) with a relapse of the disease and 15 (11.09%) with newly diagnosed hyperthyroidism. An increase of activated CD3+CD4+CD25+ was found in patients with Graves’ disease with a duration of thimazole-induced euthyroidism 5–8 months and 9–12 months, respectively, Me=0.94 (0.48–1.45), p=0.020) and Me=0.95 (0.41–1.80), p=0.025), in control group — Me=0.12 (0.03–0.68). Compared to the control an increase of CD4+CD25+CD127Low (Treg) was found in patients with a duration of thimazole-induced euthyroidism 5–8 and 9–12 months. The content of Treg in peripheral blood in Graves’ disease patients with a duration of thimazole-induced euthyroidism more than 12 months decreases, but remains elevated relative to the control.CONCLUSION: In patients with Graves’ disease with a duration of thimazole-induced euthyroidism 5–8 months and 9–12 months the level of Treg has been increased. The increase of activated Th (CD3+CD4+CD25+) persists independently of thimazole-induced euthyroidism. In patients with Graves’ disease with a duration of thimazole-induced euthyroidism for more than 12 months, there is a compensatory increase in regulatory T-lymphocyte, and the total number of T-helpers is restored to the control.

DEPENDENCE OF THE PHENOTYPIC COMPOSITION OF T-LYMPHOCYTES IN PATIENTS WITH CHRONIC VIRAL HEPATITIS C ON THE VIRUS GENOTYPE (IN DYNAMICS OF TREATMENT OF MEDICINES OF DIRECT ANTIVIRAL ACTION)

The aim of the study was to investigate the phenotype of effector T lymphocytes in patients with chronic viral hepatitis C (CVHC) in the dynamics of treatment with direct antiviral drugs depending on the genotype of the virus. 50 patients with CVHC were examined. The diagnosis was made on the basis of epidemiological and clinical laboratory data when specific serological markers of CHCV and RNA of hepatitis C virus (HCV) were detected. The determination of HCV RNA was carried out by the method of quantitative polymerase chain reaction in real time. The degree of liver fibrosis in patients with CVHC was assessed using ultrasound elastography. Patients were treated for 3 months with direct antiviral drugs according to the recommendations of the European Association for the Study of the Liver. The control group included 46 healthy donors with negative serological and molecular studies for the presence of viral hepatitis markers. The study of the subpopulation composition of helper and cytotoxic T-lymphocytes was carried out by direct immunofluorescence of whole peripheral blood. It was found that in CVHC patients were found characteristic features in the phenotypic composition of effector T-lymphocytes before and after treatment with direct antiviral drugs in depending on the genotype of HCV. The patients with HCV genotypes 1 and 3 had an increase in the content of terminal differentiated effector (TEMRA) T-helpers and effector memory (EM). Only patients with HCV genotype 2 had a decrease in the level of EM T-helper cells in the blood. A decrease in the relative number of T-helpers of central memory (СM) was independent of the HCV genotype. The level of effector subpopulations of cytotoxic T-lymphocytes in patients with CVHC was consistent with or exceeded control levels in depending on the genotype of HCV. The level of all investigated subpopulations of effector cytotoxic T-lymphocytes in patients with HCV genotype 1 is equal to the control values. The number of naive cytotoxic T cells and CM in peripheral blood in patients with HCV genotype 2 was increased. The content of naive cytotoxic T-lymphocytes, CM and TEMRA in patients with genotype 3 HCV in the blood was increased. The highest viral load was detected in patients with CVHC with genotype 1 HCV. Liver fibrosis was most pronounced in patients with CVHC infection with HCV genotypes 2 and 3. After 3 months of treatment with direct antiviral drugs the patients with CVHC had a reduced content of CM T-helpers regardless of the HCV genotype. In addition, patients with HCV genotypes 1 and 3 had a decrease in the number of naive T-helpers and patients with HCV genotypes 2 and 3 had a normalization of the content of naive cytotoxic T lymphocytes.

Effect of the combination of theophylline and budesonide on production of proinflammatory cytokines by blood cells of patients with chronic obstructive pulmonary disease

The objective: to evaluate the ability of the combination of theophylline and budesonide to suppress proinflammatory cytokine production byblood cells in patients with chronic obstructive pulmonary disease (COPD).Subjects and Methods. Peripheral blood mononuclear cells (PBMCs) or whole blood cells of COPD patients (n = 27) were incubated with budesonide (10 nM), theophylline (1 μM), or the combination thereof and stimulated with phytohemagglutinin (PHA) or phorbol myristate acetate (PMA) and ionomycin. The enzyme immunoassay was used to evaluate the secretion of thymic stromal lymphopoietin (TSLP), macrophage migration inhibitory factor (MIF), interleukin 17A (IL-17A), IL-33, and other mediators of PBMC cells, and induced PHA. The flow cytometry was used to analyze intracellular production of proinflammatory cytokines stimulated by PMA/ionomycin in T-helpers (CD4+) and cytotoxic T-lymphocytes (CD8+).Results. Theophylline reduced the secretion of IL-4 and IL-17A by PBMC cells. The combination of budesonide with theophylline inhibited the synthesis of IL-4, IL-5, IL-8, IL-13, IL-17A, IL-33, TSLP, MIF by PBMC cells as well as the production of IL-4, IL-8, tumor necrosis factor-α, and interferon-γ by cytotoxic T-lymphocytes and T-helpers. The combination of theophylline and budesonide had a more pronounced inhibitory effect on the production of IL-4 and IL-8 by PBMC cells as well as the synthesis of IL-4 by CD4+ T-cells and IL8 by CD8+ T-lymphocytes versus the effect of monotherapy with budesonide.

Peculiarities of the immune status of patients with multidrug­resistant pulmonary tuberculosis after application of treatment regimens with bedaquiline and linezolid

Objective — to investigate immuno-metabolic homeostasis in patients with new and recurrent cases of destructive forms of multidrug-resistant pulmonary tuberculosis (MDR-PTB) after treatment with bedaquiline and linezolid. Materials and methods. A clinical and laboratory examination of 175 people with MDR-PTB (89 patients with new cases and 86 patients with recurrent cases of destructive forms of MDR-PTB). The study was performed before and after treatment with bedaquiline and linezolid. The total leukocytes count (L), the leukocyte differential count (leukocyte formula), was determined in all subjects, The content of populations and subpopulations of lymphocytes were calculated using monoclonal antibodies to CD3+, CD4+, CD8+, CD19+, CD23+, CD56+ lymphocytes antigen in the reaction of indirect immunofluorescence. Quantitative determination of serum immunoglobulins was performed by Manchini radial immunodif­fusion in a gel. The level of circulating immune complexes (CIC) was determined by the spectrophoto­metric method by precipitation in polyethylene glycol. Results and discussion. In patients with new cases of destructive forms of MDR-PTB to treatment revealed hypersensitivity reactions of the first and the fourth type. Expressed activation of humoral and killer parts of immunity detected. In patients with recurrent cases of destructive forms of MDR-PTB before treatment, there are pronounced hypersensitivity reactions of the fourth type and activation of the humoral and killer parts of the immune system.Patients with new cases of destructive forms of MDR-PTB after application of treatment regimens with bedaquiline and linezolid have hypersensitivity reactions of the first and fourth types. Activation of humo­ral and killer parts of immunity detected. In patients with recurrent cases of destructive forms of MDR-PTB after the application of treatment regimens with bedaquiline and linezolid revealed a pronounced hypersensitivity reaction of the first type and activation of the killer and humoral parts of the immune system. Conclusions. In patients with new cases of destructive forms of MDR-PTB before treatment revealed T-cell immunodeficiency, which is expressed by a decrease in the level of T-lymphocytes (1.7 times relative to normal) and T-helpers (twice below normal). In patients with recurrent cases of destructive forms of MDR-PTB before treatment revealed T-cell immunodeficiency with a marked decrease in the level of T-lymphocytes (1.8 times normal) and T-helpers (1.8 times below normal). In patients with new cases of destructive forms of MDR-PTB after the application of treatment regimens with bedaquiline and linezolid revealed activation of the T-cell immune system due to increased levels of T-suppressors. In patients with recurrent cases of destructive forms of MDR-PTB after the application of treatment regimens with bedaquiline and linezolid revealed T-cell immunodeficiency due to the reduced content of T-lymphocytes (1.5 times below normal), namely T-helpers (1.9 times lower than normal).

Effect of laser therapy on the immune status of patients with chronic salpingoophoritis of chlamidial etiology

Aim. Justify the use of low-frequency laser radiation (LFLR) treating chronic salpingoophoritis of chlamydial etiology, taking into account the assessment of the immune status.Material and methods. The study included 110 women, of which 80 women had chronic salpingo-oophoritis of chlamydial etiology, and 30 were practically healthy. In addition to conventional methods, patients underwent examinations like immunological analysis, ultrasound examination of the pelvic organs, enzyme-linked immunosorbent assay (ELISA), polymerase chain reaction (PCR) analysis.Results and discussion. When comparing changes in the immune status in patients after LFLR and who underwent traditional therapy in the group of patients with sexually transmitted infections (STIs), there was a slight increase in the number of T-lymphocytes and T-helpers, as well as a decrease in the number of T-suppressors. Also, in these patients, there is a slight decrease in the number of IgA and IgM against the background of an increase in IgG and an increase in the phagocytic activity of neutrophils. In patients with traditional therapy, the above-described changes persisted, while an aggravation of the disturbed T-cell balance was observed.Conclusions. Thus, according to the results of a comparative analysis of the obtained data on changes in the immune status in patients with chronic salpingo-oophoritis after the use of laser therapy, a statistically significant increase in the number of T-lymphocytes, an increase in subpopulations including T-helpers, a decrease in the number of T-suppressors, an increase in immunoglobulin G and an increase in phagocytic activity of neutrophils was observed.

Changes in the immune status of patients with metastatic solid tumors during stereotactic radiation therapy: dependence on dose and amount of irradiated metastasis

Purpose. To evaluate immunological status in patients with metastatic forms of solid tumors before and at different intervals after stereotactic body radiation therapy (SBRT) of metastatic lesions depending on the dose and the number of irradiated metastasis. Materials and methods. A quantitative assessment and analysis of blood immunological parameters was conducted before irradiation, via 3-4 weeks and via 6-8 weeks after SBRT in patients with malignant tumors with oligometastases in the liver and lungs, in groups with a total focal dose (TFD ≤ 45 Gy ) and TFD> 45 Gy, and also in groups with irradiation of one metastases and two or more. All peripheral blood samples were analyzed by flow cytometry. Statistical analysis was performed using Friedman and Nemenyi criteria. Results. 3-4 weeks after the end of SBRT, when using higher doses (TFD> 45 Gy), we observed statistically significant increase of T-lymphocytes (CD3+CD19-); T-helpers (CD3+CD4+); activated T-helpers (CD3+CD4+HLA-DR+); activated cytotoxic T-lymphocytes (CD3+СD8+HLA-DR+). Decreasing of B-lymphocytes (CD3+CD19-) was observed in both dose groups (TFD ≤ 45 Gy and TFD > 45 Gy). We also noted the activation of the T-cell link of immunity both in the group with irradiation of one metastatic lesion, and in the group where 2 or more metastases were exposed to irradiation. Conclusion. Using of higher doses of SBRT is associated with a more activated antitumor T-cell immune response, while the analysis of groups of patients with different ammount of irradiated metastasis currently requires further research.