Effects of Levofloxacin on Blood Lymphocyte Apoptosis in Patients with Pulmonary Tuberculosis: an In Vitro Study.

The effects of a fluroquinolone levofloxacin on apoptosis of peripheral blood lymphocytes from patients with infiltrative pulmonary tuberculosis were studied in vitro. It was found that levofloxacin stimulated apoptotic cell death in tuberculosis. Addition of levofloxacin to cell suspension from patients with drug-susceptible form of tuberculosis led to an increase in the number of CD95+ and AnnV+ lymphocytes. In patients with drug-resistant form of tuberculosis, only the number of apoptotic lymphocytes, but not the count of CD95+ cells increased under these conditions.

Association between polymorphisms of cytokine genes and secretion of IL-12p70, IL-18, and IL-27 by dendritic cells in patients with pulmonary tuberculosis.

Proinflammatory cytokines are known to play a crucial role in the pathogenesis of tuberculosis, and gene polymorphisms in the promoter regions of cytokine genes were shown to substantially influence the secretory capacity of immune cells. In the present study, we analyzed the association between polymorphisms of the IL12B, IL18, and IL27 genes and the secretion of the proinflammatory cytokines IL-12р70, IL-18, and IL-27 by myeloid dendritic cells (mDCs) in pulmonary tuberculosis (PTB) patients. The study enrolled 334 patients with newly diagnosed infiltrative and disseminated PTB. Cultivation of mDCs was performed from non-proliferating progenitors of CD14+ blood monocytes. Cytokine secretion was evaluated by measuring cytokine concentration in the mDC culture supernatants using enzyme-linked immunosorbent essay (ELISA). To study cytokine gene polymorphisms, a polymerase chain reaction (PCR) and restriction fragment length polymorphism analysis (RFLP) were performed. Reduced secretion of IL-18 and IL-27 by mDCs in PTB patients was associated with 105A/C polymorphisms of the IL18 gene, and 2905T/G, 4730T/С, and -964A/G of the IL27 gene, respectively. Polymorphism IL12B/insertion had a bidirectional influence on the secretion of IL-12p70, being associated with decreased levels of the cytokine in infiltrative PTB and increased levels in disseminated PTB patients.

In Vitro Study of the Modulatory Effects of Levofloxacin and BCG on Secretion of Proinflammatory Cytokines in Infiltrative Pulmonary Tuberculosis.

The effects levofloxacin (fluoroquinolone) and vaccinal BCG strain on cytokine production by blood mononuclear leukocytes was studied in patients with infiltrative pulmonary tuberculosis. Combined treatment with levofloxacin and vaccinal BCG strain suppressed the production of TNFα in drug-resistant pulmonary tuberculosis and production of IL-12 and IFNγ in drug-sensitive tuberculosis.

Subpopulation Structure of IFNγ-Producing T Lymphocytes in Patients with Pulmonary Tuberculosis.

The study of subpopulation structure of IFNγ-producing T cells in patients with pulmonary tuberculosis revealed a decrease in the number of CD3+ IFNγ+ cells against the background of significantly increased IFNγ secretion in vitro irrespective of the clinical form of the disease and drug sensitivity of M. tuberculosis, most strongly expressed in case of the disseminated tuberculosis. In patients with infiltrative drug-sensitive and drug-resistant pulmonary tuberculosis, increased number of Th1/Th17 lymphocytes (CD4+ IFNγ+IL-17A+) and, conversely, decreased number of blood γδT cells was detected.

Expression of Galectins-1 and Galectin-3 in Stomach and Colorectal Cancer with Tissue Eosinophilia.

We analyzed the expression of galectin-1 and galectin-3 in tumor tissue in stomach and colorectal cancer with and without tissue eosinophilia. Low expression of galectin-3 was detected in all patients with malignant gastrointestinal tumors irrespective of the presence of eosinophilia. Low expression of galectin-1 was detected only in patients with gastrointestinal cancer associated with eosinophilia. Association of galectin-1 expression with eosinophilic infiltration of the tumor tissue in stomach and colorectal cancer was detected.

Tumor-associated eosinophilia.

The review provides information on current literature on structural and functional features of eosinophilic granulocytes and their role in the pathogenesis of cancer. There are examined data of clinical and experimental studies about an influence of hemic and tissue eosinophilia on the course and prognosis of malignant tumors. Molecular mechanisms of action of eosinophils in tumor pathology are discussed.

Factors of Suppression of Immune Response in Patients with Pulmonary Tuberculosis and Eosinophilia.

We studied possible mechanisms of immunosuppression mediated by regulatory T cells that promotes suppression of antigen-specific immune response to Mycobacterium tuberculosis in patients with pulmonary tuberculosis and eosinophilia. It was shown that the number of CD4(+)CD25(+)Foxp3(+) regulatory T cells with immunosuppressive activity (Treg) increased in the peripheral blood of patients with disseminated destructive forms of pulmonary tuberculosis with multiple resistance of the causative agent to antituberculosis substances and eosinophilia. These changes were accompanied by imbalance in secretion of Treg-associated cytokines (in vitro) manifested in hyperproduction of TGFß and IL-10 and decreased production of IL-2.

Molecular Factors of Cytokine-Dependent Activation of T Cells in Pulmonary Tuberculosis.

The parameters characterizing antituberculous immune response at the stage of cytokinedependent activation of T cells were analyzed in 90 patients with pulmonary tuberculosis before etiotropic therapy. Immunophenotyping of T cells for IL-12Rß2, WSX-1, and gp130 surface markers and T-bet intracellular transcription factor was carried out after specific IL-12/IL-27 induction of cells in vitro. An increase in the counts of CD3(+)T-bet(+), CD3(+)IL-12Rß2 (+), and CD3(+)WSX-1(+)gp130(+) cells and the level of T cells with high expression of WSX-1 inhibitory molecule (CD3(+)WSX-1(hi+)gp130(-) and CD3(+)WSX-1(hi+)gp130(+)) was detected. The type of disorders in the inductive stage of antituberculous immune response did not depend on the clinical form of disease.

[THE INDICATORS OF COAGULATION HEMOSTASIS AFTER CRIO-RESECTION OF LIVER].

The article presents results of comparative evaluation of indicators of coagulation hemostasis in 24 patients with focal parasitic and non-parasitic pathology of liver before and after resection of organ with and without frigotherapy. The analysis of condition of coagulation hemostasis was implemented on the basis of results of blood clotting tests and immune enzyme technique of detection of content of factors V, Xl, XII in blood plasma. It is demonstrated that after crio-resection of liver the tendency to normalization of content of factors V and XI is more expressed than in case of common resection of organ. At that, in patients with non-parasitic diseases of liver after crio-resection initially decreased concentration of factor Xl is normalized and content of factor XII keeps to be in limits of normality under its decreasing in patients after resection without frigotherapy. The plasma concentration of fibrinogen corresponds to standard. The most significant departures of international normalized ratio (increasing higher than reference values) and activated partial thromboplastin time (decreasing up to reference values) in patients with parasitic and non-parasitic diseases of liver are observed at 5th day after resection independently of technique applied (with of without frigotherapy).

Factors of Th17 and Treg Lymphocyte Differentiation in Pulmonary Tuberculosis.

In patients with infiltrative pulmonary tuberculosis, the increase in IL-6 secretion, the decrease in TGFß production (in case of drug resistance of the causative agent), and unchanged level of IL-1ß secretion by mononuclear leukocytes in vitro were associated with increased number of CD4(+)CD161(+)IL-17A(+) Th17 lymphocytes in the peripheral blood. In patients with disseminated drug-resistant pulmonary tuberculosis, TGFß hyperproduction promoted differentiation of CD4(+)CD25(+)FoxP3(+) Treg lymphocytes with immunosuppressive activity.

[Expression of mRNA transcription factors RORC2 and FoxP3 in lymphocytes in patients with pulmonary tuberculosis].

Homeostasis of subpopulations Th17- and Treg-lymphocytes plays an important role in a holistic and coordinated process of eradication of pathogens and preventing the spread of infection in the body. Study of molecular mechanisms controlling the balance of these cells in the formation of immune deviation in the pathogenesis of infection are particularly relevant. The article presents the results of a study of mRNA expression of transcription factors Th17- and Treg-lymphocytes--RORC2 and FoxP3, respectively, as well as the presence of these cells in peripheral blood in infectious disease (based on an example of infection caused by Mycobacterium tuberculosis). It was established that during the infiltrative (regardless of drug susceptibility testing) and disseminated drug-susceptible pulmonary tuberculosis accompanied by Th17-polarized differentiation of T-lymphocytes, as evidenced by the increased number of CD4+CD161+IL17A+ cells in the blood in association with increased mRNA expression of the transcription factor RORC2 in lymphocytes. In disseminated drug-resistant pulmonary tuberculosis T-lymphocyte differentiation is carried out mainly in the direction of immunosuppressive Treg-cells, as evidenced by the increase in their number in the blood in association with elevated levels of mRNA expression of the transcription factor FoxP3 in lymphocytes.

Functional activity of Th-17 lymphocytes in pulmonary tuberculosis.

Increased content of CD4(+)CD161(+)IL-17A(+) Th17 lymphocytes in the peripheral blood was found in patients with pulmonary tuberculosis irrespective of the clinical form (infiltrative, disseminated) and variant of the disease (drug-sensitive, drug-resistant). The elevated content of Th17 cells in pulmonary tuberculosis is associated with hypersecretion of Th17-associated cytokines IL-17A and IL-22 in vitro that was most pronounced (in case of IL-17A) in patients with disseminated pulmonary tuberculosis.

[Effect of eprosartan on the hemostatic system in patients with chronic kidney disease associated with hereditary thrombophilia].

AIM: To study the effect of eprosartan, an angiotensin II type 1 (AT1) receptor blocker, with sympatholytic activity on the hemostatic system in patients with chronic kidney disease (CKD) associated with hereditary thrombophilia. SUBJECTS AND METHODS: The 12-week open-label uncontrolled trial included 31 patients with Stages I-II CKD: 15 patients with chronic glomerulonephritis and 16 with diabetic nephropathy burdening types 1 and 2 diabetes mellitus (DM) in 10 and 6 cases, respectively. In all the patients, CKD was associated with one of the heterozygous forms of thrombophilia: the polymorphic methylenetetrahydrofolate reductase gene variant C677T was found in 18 patients; the polymorphic coagulation factor V gene variant G1691A was in 9; and the polymorphic coagulation factor II gene variant G20210A in 4. Along with the thorough examination accepted in nephrology and endocrinology clinics, investigations of vascular-thrombocytic and secondary hemostasis and the anticoagulant and fibrinolytic systems were made before and after treatment. RESULTS: Eprosartan therapy caused positive changes in the indicators of vascular-thrombocytic (diminished platelet aggregation, reduced surplus of von Willebrand factor and endothelin-1) and secondary (decreased coagulation factor VII activity, longer activated partial thromboplastin time) hemostasis and the anticoagulant (reduced antithrombin III deficiency) and fibrinolytic (elevated blood plasminogen concentrations) systems. CONCLUSION: The pleiotropic effects of eprosartan may be used to correct hypercoagulability syndrome in patients with CKD associated with hereditary thrombophilia.

[The relationship of AB0- and rhesus-phenotypes of erythrocytes with expression of intra-operational hemolysis in cardio-surgical patients].

The study sampling included patients with ischemic heart disease with mild (70 patients) and marked (36 patients) hemolysis after coronary artery bypass grafting under artificial blood circulation. During post-operation period the content of free hemoglobin in blood plasma, AB0- and rhesus-phenotype of erythrocytes were evaluated. It is established that in patients with marked intra-operational hemolysis as compared with cases of mild hemolysis the phenotypes of erythrocytes B(III), AB(IV), ccDEE, ccDEe are found reliably more often and 0(I)-phenotype is found reliably more rare. The risk factor of marked intra-operational hemolysis is a verification of ccD(E/e)-phenotype of erythrocytes and in case of different rhesus-phenotypes--blood type B(III) or AB(IV).

[Characteristic of the complement system in patients with ischemic heart disease with moderate and marked hemolysis after operations with cardiopulmonary bypass].

A study of the complement system in cardiosurgical patients with moderate (40 patients) and marked (18 patients) hemolysis after coronary artery bypass grafting in conditions of cardiopulmonary bypass was carried out. Before and after operation the content of D35+-, D55+-erythrocytes and reticulocytes in blood, free hemoglobin in blood plasma, indicators of the functional state of classical, lectin and alternative pathways of complement activation as well as concentration of its terminal complex in blood serum were analyzed. It was established that development of marked hemolysis was associated with higher (compared with moderate hemolysis) content of terminal complement complex and reticulocytes in blood before operation as well as deficiency of D55+- erythrocytes and low activity of alternative pathway.

[T-lymphocyte-helper type 17 mediated regulation of antibacterial (antituberculosis) immunity].

The analysis of current views on functional and immunoregulatory role of T-lymphocytes-helpers of type 17 (Th17) in anti-infectious immune response is presented, in particular, in the development of protective immune reactions to intracellular pathogens. Particular attention is paid to participation of these lymphocytes and cytokines produced by them in immune response to Mycobacterium tuberculosis invasion. The molecular mechanisms, underlying the predominant development of Th17-lymphocytes and/or regulatory T-cells (Treg), are studied, with evaluation of interconnection of these cell subpopulations in formation of immune imbalance in infectious pathology.

[The biochemical markers of endothelium dysfunction in patients with diabetic nephropathy].

The article deals with studying the degree of increase of the von Willebrand factor and the concentration of endothelin-1 in blood plasma in the subgroups of patients with diabetes mellitus formed depending on of type of disease and presence of phenotype with affection of kidneys. The sampling of 176 patients with diabetes mellitus (65 patients with diabetes mellitus type 1, 111 patients with diabetes mellitus type II) was examined. The control group consisted of 30 healthy persons. In the capacity of biochemical markers of endothelium dysfunction the activity of the von Willebrand factor and the concentration of endothelin-1 in blood were considered. In all patients with diabetes mellitus the biochemical characteristics of endothelium dysfunction are present manifesting by increase of concentration of endothelin-1 in blood which is especially expressed under disease phenotype with affection of kidneys. Despite of the apparent lesion of endothelium in patients with diabetes mellitus compromised with diabetic nephropathy the thrombocytes aggregation induced by ristomicine does not undergo natural changes. Hence, to consider in these patients the increasing activity of the von Willebrand factor as a reliable marker of endothelium dysfunction is not seemed possible.

[Factors of intravascular hemolysis in cardiosurgical patients after cardiopulmonary bypass procedures].

The study included patients with ischemic heart disease with moderate (52 patients) and apparent (23 patients) hemolysis after coronary bypass surgery in cardiopulmonary bypass (CB). The concentration of free hemoglobin in blood plasma, mechanical resistance and sorption capacity of red cells as well as the content of TBA-active products, cholesterol and phospholipids in red cells and reticulocytes levels in blood were studied before and after operation. It was shown that among patients with apparent post-perfusion hemolysis (in contrast to the patients with a moderate hemolysis) the sorption capacity of red cells and amount of reticulocytes in blood are increased before operation; level of TBA-active products in erythrocytes is increasing after operation. Development of moderate hemolysis is associated with the decreased mechanical resistance of erythrocytes and increased cholesterol/phospholipid-ratio in membranes before operation. Thus, individually-specified apparent post-perfusion hemolysis is based on free-radical mechanism of erythrocytes damage and moderate hemoglobin level is referred to mechanical trauma of blood cells during CB.

[Molecular mechanisms of formation of blood eosinophilia under pulmonary tuberculosis].

Molecular factors of pathogenesis of the eosinophilic blood reaction under pulmonary tuberculosis are analyzed in the article. It has been established that the key cytokine providing the development of hemic eosinophilia in patients with pulmonary tuberculosis is IL-5. IL-5 plasma concentration turned out to be increased only in patients with eosinophilia. Increase of eotaxin was determined in patients with tuberculosis despite of the presense of eosinophilia. One-directional nature of the defined changes in eotaxin concentration might be explained by dual properties of this chemokine: on the one hand, eotaxin mediates long-term presence of eosinophils in blood; on the other hand, it initiates the process of adhesion of eosinophilic leucocytes to vascular endothelium with their further migration to the focus of granulomatous inflammation. The established increase in number of IL-5R-positive eosinophils presents one more mechanism which explains the basis of long-term presence of eosinophils in peripheral blood in patients with pulmonary tuberculosis.

Cytokine-secreting activity of blood eosinophils in pulmonary tuberculosis.

Modern immunological studies showed that eosinophilic granulocytes producing the key mediators of cellular and humoral immune response contribute to the common cytokine imbalance developing in tuberculous infection. A significant increase in BCG-induced secretion of IL-2, IL-5, and TNF-α by eosinophils in patients with pulmonary tuberculosis indicated high reserve reactivity of eosinophilic cells realizing their functional potential in regulation of the specific resistance reactions of the microorganism under conditions of M. tuberculosis infection.