Functional Phenotype of Alveolar Macrophages Features in Rats with Metabolic Syndrome.

Phenotypic characteristics of alveolar macrophages in the bronchoalveolar lavage fluid as well as their ability to acquire the M1 and M2 phenotypes during in vitro culturing with reprogramming factors were studied in rats with modeled diet-induced metabolic syndrome. A decrease in the number of alveolar macrophages with the M1 phenotype was found in animals with metabolic syndrome. The factors of metabolic syndrome do not affect phenotypic plasticity of cells in culture, but under the action of M2 reprogramming factors, the cells demonstrate a wide range of phenotypic plasticity by the CD80 and CD206 markers. The consistently high level of production of IL-6 and IL-10 by macrophages during culturing under different conditions indicates functional rigidity of the cells, which is probably a consequence of in vivo predetermined functional phenotype of these cells against the background of metabolic disorders.

Airway Smooth Muscles Contractions in Metabolic Syndrome.

We studied contractile responses of isolated airway smooth muscle segments from rats with metabolic syndrome. Metabolic syndrome was induced in rats by high-fat and high-carbohydrate diet. It was shown that metabolic syndrome was associated with an increase of bronchoconstrictor action of cholinergic receptor activator carbacholine (0.1-100 µM) and a decrease of the dilatory effect of ß2-adrenoreceptor activator salbutamol (0.1-100 µM). The observed effects of agonists are epithelium-dependent. Disorders in contractile activity in the airway smooth muscles were accompanied by bronchial epithelium destruction, immune inflammation in the bronchial wall, muscular and peribronchial adipose tissue hypertrophy.

The interaction effect of angiogenesis and endothelial dysfunction-related gene variants increases the susceptibility of recurrent pregnancy loss.

PURPOSE: The role of genetic polymorphisms in the pathogenesis of recurrent pregnancy loss (RPL) has been studied intensively. Complex diseases, including miscarriage, are believed to have a polygenic basis, and gene-gene interactions can play a significant role in the etiology of the disease. This study was conducted to investigate the association of gene-gene interactions with angiogenesis, endothelial dysfunction-related gene polymorphisms, and RPL. METHODS: A case-control study was conducted with 253 unrelated RPL patients with 2 or more spontaneous pregnancy losses and 339 healthy women with no history of pregnancy complications. Genotyping of single-nucleotide polymorphisms (SNPs) was performed using real-time polymerase chain reaction (real-time PCR), restriction fragment length polymorphism (RFLP), or allele-specific polymerase chain reaction methods. RESULTS: The genotypes 677TT of the MTHFR gene, 936TT, 936CT, and 634CC, 634GC of the VEGF gene, and allele 894T of the NOS3 gene were associated with a predisposition to RPL in the Russian population. A significant role of additive and epistatic effects in the gene-gene interactions of the SNPs of SERPINE-1, ACE, NOS3, MTHFR, and VEGF genes in RPL was demonstrated. CONCLUSIONS: The results showed that gene-gene interactions are important for RPL susceptibility. Additionally, analysis of the genotype combinations of several allelic variants provides more information on RPL risk than analysis of independent polymorphic markers.

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.

[The immune stressor effects of T-regulatory cells under infiltrative tuberculosis of lungs].

The article deals with the characteristics of sup-population composition of T-regulator: cells with suppressor activity and production of immunoregulatory cytokines suppressing immune response (IL-10, TGF-beta) in patients with infiltrative tuberculosis of lungs. It is proved that the leading role in formation of immunodepression and tuberculin anergy under infiltrative tuberculosis of lungs is played by T-regulatory, cells with phenotype CD4+CD25+Foxp3+. It is demonstrated that the immunodepression mediated by cytokine production is connected with increasing of both basal and BCG-induced secretion of IL-10 on the background of decrease of level of production of TGF-beta by mononuclear leukocytes in vitro.

[Subpopulation structure of T-regulatory cells and proliferative response of blood lymphocytes in vitro under pulmonary tuberculosis].

Subpopulation structure of T-regulatory cells and proliferative activity of blood lymphocytes in vitro in patients with different clinical forms of pulmonary tuberculosis were studied in this work. It has been shown that Trn--natural T-regulatory lymphocytes (CD4+CD25+Foxp3+) play a leading role in formation of immune suppression under infiltrative, disseminated and fibrosis-cavernous pulmonary tuberculosis. Besides, their number is increased in blood of both tuberculin-positive and tuberculin-negative patients. Negative correlation between the number of Trn and proliferative activity of blood lymphocytes (basal, mitogen- and antigen-induced)has been established, which testifies about participation of Trn in suppression of lymphocyte proliferation and Th1- and Th2-immune response.

Allelic polymorphism of cytokine genes during pulmonary tuberculosis.

Modern immunological and molecular genetic studies showed that tuberculosis is accompanied by an imbalance in the production of immunoregulatory cytokines by mononuclear leukocytes. T allele and homozygous TT genotype of T-330G polymorphism in the IL2 gene, T allele and TT genotype of C-590T polymorphism in the IL4 gene, and CC genotype of A-1188C polymorphism in the IL12B gene are immunogenetic factors that have protective activity against susceptibility to pulmonary tuberculosis. Susceptibility to tuberculous infection is associated with A1A2 genotype of the polymorphic region +3953 A1/A2 in the IL1B gene; G allele and TG and GG genotypes of T-330G polymorphism in the IL2 gene; C allele and CC and CT genotypes of C-590T polymorphism in the IL4 gene; and AC genotype of the polymorphic region A-1188C in the IL12 gene.

[Allelic polymorphism of the IL-2and IL-4 genes in herpesvirus infection].

Herpesvirus infection is accompanied by imbalance of peripheral mononuclear production of cytokines, by polarizing an immune response by the Th2 pathway. The homozygous TT genotype of T330G polymorphism in the IL-2 gene is an immunogenetic factor that has a protective activity against susceptibility to herpesvirus infection. The risk of progression, the recurrent course of the infection and its poor outcome is positively associated with the heterozygous TG genotype in the IL-2 gene T-330G promoter region and with the homozygous CC type and C allele in the IL-4 gene C-590T promoter region.

[Modulatory effect of isoniazid and rifampicin in vitro on cytokine secretion in pulmonary tuberculosis].

The effects of isoniazid and rifampicin on the peripheral mononuclear production of IL-2, IL-12, IL-10, IFN-gamma, and TGF-beta were evaluated in patients with infiltrative pulmonary tuberculosis. Irrespective of the susceptibility of the causative agent to the essential antituberculous drugs, rifampicin was ascertained to initiate increased IL-2 secretion and to reduce the generation of IL-12, IFN-gamma and TGF-beta. Isoniazid suppressed the mononuclear leukocytic production of IFN-gamma in drug-sensitive pulmonary tuberculosis, and conversely, stimulated it in the drug-resistant type. Rifampicin showed a more significant negative effect on mononuclear cytokine-producing activity. In drug-resistant pulmonary tuberculosis, the imbalance of spontaneous, protein- and drug-induced mononuclear secretion of cytokines was established to be more pronounced than in drug-sensitive tuberculosis.

[The molecular-genetic aspects of tubercular infection forecasting and immunotherapy].

In article is presented, the analysis of the data of the literature and own researches, concerning molecular-genetic mechanisms of antimycobacterial immunity infringements. The role of the factors defining features of a current and an outcome of a tubercular infection, namely of immunity system condition, feature of infecting microbic Mycobacterium tuberculosis strain biological properties (a genotype, a spectrum of drag sensitivity/resistance), influence of combined antitubercular chemotherapy is disc issed. The opinion is expressed that the given factors are necessary for taking into consideration for working out methodology the personalised preventive maintenance and correction of immunity infringements during pulmonary tuberculosis.

[Apoptosis and micro- and macroelement composition of lymphocytes in patients with pulmonary tuberculosis].

The paper describes the results of comparatively analyzing the parameters of apoptosis, micro- and macroelement spectrum of peripheral blood lymphocytes in healthy donors and in patients with chronic obstructive pulmonary disease (COPD) and pulmonary tuberculosis. In drug-responsive and drug-resistant pulmonary tuberculosis, unidirectional pretreatment changes were generally recorded in the activity of apoptosis and the spectrum of chemical elements of peripheral blood lymphocytes, which were most pronounced after completion of a phase of intensive antituberculosis chemotherapy. After termination of a complete course of the therapy, apoptotic activity lowered with the cationic composition of cells being normalized. The parameters differentiating pulmonary tuberculosis from COPD were ascertained, these included increases in the count of apoptotic cells and the cell concentrations of Ca2+, Zn2+, and Mg2+.

[Multidrug pulmonary tuberculosis: sociomedical features and the efficiency of inpatient treatment].

Case histories of 147 patients with new-onset destructive pulmonary tuberculosis (PT) (infiltrative, disseminated, fibrocavernous) with the pathogen of the disease showing multidrug resistance (MDR) were analyzed, by evaluating the efficiency of treatment involving sputum abacillation after termination of the inpatient stage of treatment. A control group included 220 patients with PT of the similar lesion and clinical forms, who preserved drug resistance (DR) to antibacterial agents. Most patients with MDR disseminated destructive PT are young people aged 36.6 +/- 1.6 years, this disease-associated disabled individuals (65.3%), alcoholics (48.3%), opium addicts (11.5%), ex-prisoners (26.5%), single, homeless; more frequently suffer from gastrointestinal (40%) and chronic nonspecific lung (24.5%) diseases. The course of MDR PT is significantly more commonly complicated by the development of respiratory failure (48.9%) and hemoptysis (6.1%) (in DR PT 19.1 and 2.3%, respectively). In terms of negative smear tests and the results of sputum cultures, the abacillation rates were 70.1 and 67.3%, respectively (in DR PT, these were 88.18 and 86.34%, p = 0.01). A negative reaction of sputum occurred in 42.1% of patients at 2-3 months of treatment while its culture did in 46.2% at 4-5 months. In the control group wherein drug sensitivity of M. tuberculosis was preserved, sputum abacillation occurred in the larger proportion of the patients within the first 2 months (in 68.6 and 56.3% of cases with sputum smear and culture, respectively). The efficiency of inpatient therapy is greatly affected by short-term treatment caused by voluntary withdrawal and irregular uses of antituberculous drugs, mainly due to alcoholization.

[Evaluation of the effects of antituberculous drugs on the cytochemical status of lymphocytes in vitro].

The effects of basic drugs of chemotherapy for tuberclosis, such as isoniazid and rifampicin, on the activity of the most important mitochondrial enzymes of lymphocytic energy metabolism were studied in patients with infiltrative tuberculosis and healthy donors. The authors detected the significantly inhibited activity of succinate dehydrogenase and the redistribution of lymphocytic populations with the increased proportion of cells with the low and high activities of the enzyme. The more significant negative effect on the lymphocytes of patients with tuberculosis was observed when rifampicin was administered. In patients with pulmonary tuberculosis, the suppressed enzymatic activity of peripheral blood lymphocytes was shown to be more pronounced when the cells were incubated with rifampicin and isoniazid for 2 hours than for 1 hour.

[Role of cytokines in the modulation of lymphocytic subpopulational composition in patients with pulmonary tuberculosis].

The parameters of the lymphocytic CD-population composition and the production of cytokines in the cell cultures of peripheral blood mononuclear cells were studied in patients with infiltrative pulmonary tuberculosis. A marked reduction in the relative and absolute count of CD3+ and CD4+ lymphocytes and an increase in the level of CD8- and CD16-positive cells were established in both drug-sensitive and drug-resistant types of the disease. A significant increase in the production of IL-12 and a reduction in the spontaneous elaboration of interferon-gamma were observed in all the examinees. Addition of lipid and protein myobacterial antigens (Beijing strains) caused a notable decrease in the generation of the study cytokines as compared with the respective parameters in healthy donors and their basal secretion irrespective of the type of a tuberculous process.

[Immune pathology in pulmonary tuberculosis].

The data of modem literature and the results of original investigations on mechanisms of immunopathological alterations in tuberculosis infection are presented. The role of cellular and humoral parts of immunity in pathogenesis of pulmonary tuberculosis is discussed and cytokine-mediated mechanisms of disorders in a specific immune response are analysed.

[The superficial architectonics of peripheral blood lymphocytes in patients with pulmonary tuberculosis].

Peripheral blood lymphocytes were electron microscopically studied in patients with disseminated drug-sensitive and drug-resistant pulmonary tuberculosis before and during specific antituberculous chemotherapy. There was a tendency for an increase in the count of lymphocytes with membranous morphostructural changes during the therapy, which was more clearly defined in the drug-resistant type of the infectious process. It is suggested that antituberculous drugs have a damaging effect on the membrane of immunocompetent blood cells.

[Evaluation of biochips for the rifampin resistance detection of M. tuberculosis in strains isolated at the Novosibirsk and Tomsk regions].

Recently in Russia biochips for rifampin resistance detection of M. tuberculosis were developed. To investigate the conformity between rifampin resistance results determined both by the routinely used absolute concentration method and USING the biochips, 272 DNA samples of M. tuberculosis isolated from TB patients at Novosibirsk and Tomsk regions in 2000-2005 were analyzed. The biochip can detect 30 mutations in rpoB gene. The mutations were also tested using the single stranded conformational polymorphism method (SSCP). In addition, 60 DNAs were randomly sampled and sequenced. The results of rifampin resistance detection using biochip and absolute concentration methods were congruent in 86% cases, and were different when analyzed samples consisted of the susceptible and resistant strains of M. tuberculosis mixture. The most frequent mutations in the rpoB gene were S531 (76.2%), H526 (7%), D516 (5.6%), and L511 (5.6%). In 94% of rifampin resistant strains, there was also resistance to isoniazid. Therefore, in Siberia the rifampin resistance is the reliable marker for MDR strains of M. tuberculosis, and biochips can be used also for their detection. To hybridize with biochip the fluorescent-labeled single-stranded DNAs were routinely synthesized by two PCR, and intermediary product after the first PCR should be transferred into another tube. The last stage included high risk of cross-contamination. To exclude the risk, primer concentrations and temperature-time profile of PCR reactions were improved, and both PCR were combined in one tube. The two methods were congruent in 100%. The one tube method would be especially attractive for the routine PCR laboratory.

[Functional activity of phagocyte blood cells in pulmonary tuberculosis].

The characteristics of the functional activity of phagocyte blood cells in patients with destructive pulmonary tuberculosis caused by medicinal sensitive and medicinal resistant infective agents were studied. In the process of pulmonary tuberculosis, irrespective of the medicinal sensitivity of infective agents before and during treatment, a decrease in the phagocytic activity of neutrophil granulocytes and the level of the expression of Fcgamma- and C3b-receptors on monocytes with a simultaneous increase in the spontaneous production of oxygen metabolites in neutophils and a rise in the adsorptive capacity of monocytic cells were observed.

[Peripheral blood mononuclear leucocytes in patients with drug-sensitive and drug-resistant pulmonary tuberculosis].

The authors studied functional characteristics of mononuclear leucocytes in patients with drug-sensitive and drug-resistant pulmonary tuberculosis. The study found decrease of the number of C3b- and Fcchi-receptorpositive monocytes and increase of their saturation activity in both categories of pulmonary tuberculosis before and after antituberculous therapy. The study revealed increase of interferons alpha and chi production and, at the same time, decrease of tumor necrosis factor alpha, which was more prominent in cases of drug-sensitive tuberculosis, and depression of interleukin 2 secretion, more prominent during chemotherapy in cases of drug-resistant tuberculosis.