Features of the functional activity of macrophage link of immunity with gastroesophageal reflux disease depending on the type of reluctate: in vitro model.

AIM: A generalized analysis of changes in functional activity of macrophages on the basis of phagocytic activity, cytokine profile, changes in the level of expression of surface markers characteristic of pro- or anti-inflammatory phenotype of the cells when exposed to reluctate. MATERIALS AND METHODS: Developed in vitro model of co-peritoneal macrophages of mice With57/BL6 (n=65) and reluctate patients with gastroesophageal reflux disease (GERD; n=65) having different pH values (three group comparison). Took into account the standard criteria phagocytic ability (absorption Staphylococcus aureus 9198, light microscopy), secretory activity (cytokine profile Th1/Th2, flow cytometry) and receptor characterization of macrophages (expression of CD25/80/163/206, flow cytometry). RESULTS: The phagocytic activity of macrophages, calculated on the basis of the average number of bacteria ingested by one phagocyte, is not associated with the pH value of the added reluctate. It is established that the alkalinisation of reluctate leads to significant alteration in the expression of CD receptors - decrease M1 and increase M2. The index of total production of Th1/Тһ2 in groups progressively decreased with increasing pH of reluctate and amounted to 3.6 units in the group pH from 4.6 to 6.6; 2.8 units group a pH of 6.7-7.2 and 1.6 units in the group pH of 7.3 to 8.1, due to increased production of Th2 cytokines at offset reluctate pH to slightly alkaline side. The data obtained indicate the increase of expression and secretion of anti-inflammatory markers at an alkaline pH shift of reluctate. Analysis of the studied characteristics of the activity profile of macrophages in the proposed in vitro model justifies the need for considering the peculiarities of the functional activity of macrophages under the influence of reluctate different nature. The special importance of studying the cytokine profile and characteristics of the functional activity of macrophages in patients with GERD, given the nature of reluctate.

[Reprogrammed M1 macrophages with inhibited STAT3, STAT6 and/or SMAD3 extends lifespan of mice with experimental carcinoma].

Objective. Reprogramming of M1 macrophage phenotype with inhibited M2 phenotype transcription factors, such as STAT3, STAT6 and SMAD and assess their impact on the development of Ehrlich carcinoma (EC) in vitro and in vivo. Methods. Tumor growth in vitro was initiated by addition of EC cells in RPMI-1640 culture medium and in vivo by intraperitoneal of EC cell injection into mice. Results. It was found that M1-STAT3/6- SMAD3 macrophages have a pronounced anti-tumor effect in vitro, and in vivo, which was greater than anti-tumor effects of M1, M1-STAT 3/6, M1-SMAD3 macrophages and cisplatin. Conclusion. M1 macrophages with inhibited STAT3, STAT6 and/or SMAD3 effectively restrict tumor growth. The findings justify the development of new anti-tumor cell therapy technology.

[Genetic features of nitric oxide generating systems predetermine the body's resistance to the development of carcinoma].

Predisposition to tumors is often determined by how effectively the genotype of an individual forms an immune defense. An important factor of such protection is macrophage NO. We assumed that the body's vulnerability to the development of tumors may depend from the characteristics of the NO generating systems. The content of NO in the tumor changed by ITU, inhibitor of iNOS, c-PTIO, traps and SNP, donor NO. Production of macrophage NO were evaluated by nitrites in the culture media. iNOS was assessed using the Western blot analysis. Phenotype of macrophages was assessed using cytometry for CD labels. Life span of mice C57BL/6N with Ehrlich tumor was 25% greater than that of the C57BL/6J. Reducing the content of NO in the tumor reduced life expectancy of high-resistance to tumor subline C57BL/6N at 23%. Increase of NO increased life expectancy of low-resistance subline C57BL/6J at 26%. Macrophages of C57BL/6N were 1.5 times higher contents of iNOS and NO production, as compared with macrophages of C57BL/6J. CD phenotype markers determined the macrophage phenotype C57BL/6N as M1 and C57BL/6J mice macrophage phenotype as M2. Thus, the body's vulnerability to the development of tumors may depend from the characteristics of the NO generating systems. C57BL/6J, unlike C57BL/6N does not synthesize NNT (nicotinamide nucleotide transhydrogenase) and have differences in the single nucleotide polymorphism (SNP). The important role of NO in the resistance to Carcinoma, NNT and SNP deserve attention in the development of new methods of antitumor therapy.

[The role of genetic peculiarityes (of organisms) in the resistance to neoplastic processes in August line and Wistar population rats].

The probability of development of the Ehrlich's ascites carcinoma in young August and Wistar rats was investigated. The Ehrlich's carcinoma strain was derived in mice in the N.N. Blokhin Russian Cancer Research Center. The tumor was transplanted into rats intraperitonially. It was shown that the transplanted murine carcinomas did not arouse tumors in rats, but caused pathologic effects: abrupt growth impairment and partial loss in the August rats while in the Wistar rats the growth impairment was slight and there was no loss. Thus, the first, there was no tumor growth in rats and the second, the indicated effects of the murine tumor transplantation were more dramatic in the August rats than thouse in the Wistar rats.

[The total content and oligomeric transformations of surfactant protein d in bronchoalveolar lavage fluid in bronchial asthma and gastroesophageal reflux disease: the role in deterioration of the immune response].

The study of pathogenesis of bronchial asthma (BA) and gastroesophageal reflux disease (GERD) or their combination showed that the intensity of inflammation and the choice between Thl and Th2 immune responses are determined by macrophages (elements of congenital immunity). Lung surfactant protein D (SP-D) existing in various oligomneric forms (as monomer trimer, dodecamer, multimer) plays an important role in the mechanism of transformation ofalveolar macrophage phenotype. Patients with BA+GERD have higher SP-D level in the bronchoalveolar lavage fluid than those with GERD alone but lower than patients with BA. SP-D dodecamers were found only in BA patients given basaLtherapy with inhaled glucocorticoids (IGC). It suggests that the presence of dodecamers in the lavage fluid may result firom anti-inflammatory action of IGC. They are absent in patients with BA+GERD treated with IGC probably because GERD enhances inflammatoly changes in the lungs of BA patients despite basal therapy These data together with results of experimental acidification of lavage fluid from BA patients give reason to hypothesize that microaspiration of acidic gastric contents frequently associated with GERD is a cause of local decrease of pH in different segments of the bronchial tree triggering two pathogenetic mechanisms: (I) programming proinflammatory MI phenotype of alveolar macrophages, increased production of nitric oxide, nitrosation of SP-D and destruction of its anti-inflammatory multimers ; (b) direct destruction ofSP-D oligomers in the acid medium. Both mechanisms reduce the level of anti-inflammatory SP-D multimers and increase the level ofproinflammatory monomers. Thus, decreased pH in lower airways is a real pathogenetic factor of anti-inflammatory shift in the oligomeric SP-D composition accounting for the inflammatory reaction of lungs in GERD.

[Functional activity of alveolar macrophages in patients with bronchial asthma and gastroesophageal reflux disease].

Combination of bronchial asthma (BA) and gastroesophageal reflux disease (GERD) is a widespread clinical situation. The two pathologies are known to influence each other leading to disturbances in immune responsiveness. We studied phenotypes and phenotypic plasticity of immune cells (alveolar macrophages) in patients with BA and GERD. It was shown that BA and GERD are largely associated with AM of proinflammatory M2 and anti-inflammatory M1 phenotypes respectively. Population of AM with MI phenotype increases in patients having both BA and GERD compared with that in BA alone. In vitro experiments showed that acidic milieu promotes shifting the phenotype toward the predominance of M1, i.e. simulates the situation characteristic of GERD. Combination of BA and GERD narrows the interval within which AM can change MI phenotype (i.e. makes them more "rigid") but broadens the range in which they can change M2 phenotype. Also, GERD promotes the development of morphological rigidity of AM. Patients with BA given steroid therapy undergo inversion of phenotypic plasticity of AM. These data characterize the immunological component of BA and/or GERD pathogenesis. They help to better understand mechanisms of development of broncho-pulmonary pathology in GERD patients and can be used to work out new methods for the treatment of these diseases.

[Resistance to acute hypoxia and changes in the phenotype and phenotypic plasticity of macrophages in mice of different genetic strains].

The aim of study was to investigate the effect of hypoxia on the macrophage phenotype and phenotypic plasticity and to determine the resistance to acute hypoxia in C57/BL mice, which have the pro-inflammatory M1 macrophage phenotype, and in BALB/c mice, which have the anti-inflammatory M2 macrophage phenotype. The following results were obtained. 1) The response of macrophages to acute hypoxia has two successive phases, the immediate, anti-inflammatory phase, and the delayed, pro-inflammatory phase. This response was more distinctly inverted in C57/BL6 M1 macrophages than in BALB/c M2 macrophages; 2) the effect of acute hypoxia on macrophage phenotypic plasticity depends on the genetically predetermined, original macrophage phenotype. In this process, a clear regularity was observed: hypoxia increased the capability of macrophages for changing into the pro-inflammatory M1 phenotype, while their capability for changing into the anti-inflammatory M2 phenotype remained virtually unaffected. 3) BALB/c mice were more resistant to acute hypoxia than C57/BL6 mice. Taken together, these data expand our understanding of mechanisms for pathogenetic effects of hypoxia.