Tumour adaptation to immune factors: old and new ideas for cancer immunotherapy.

The tumour is fully functional in the zone of action of immune mediators. Moreover, the tumour needs immune system mediators to survive. "Adaptation" refers to a tumour's ability to withstand the effect of harmful elements. This gives birth to a new form of antitumour therapy: blocking tumour adaptability pathways. In this review, we will look at (i) tumour adaptation mechanisms as a result of pro-tumour immunoediting, (ii) how understanding tumour-adaptive mechanisms has led to ideas for developing cancer immunotherapies, and (iii) prospects for using the adaptation theory to substantiate new approaches to tumour growth inhibition. By considering the cancer problem through the lens of adaptability, a unique strategy for enhancing the efficacy of immunotherapy was proposed. The new approach is to utilise antisense treatment to erase the structural trace of adaptation in tumour cells or to disadapt tumour cells by "turning off" the immune system before initiating immunotherapy.

[Growing of murine tooth in situ by homotopic transplantation of embryonic germ]. / Vyrashchivanie zuba myshi in situ metodom gomotopicheskoi transplantatsii émbrional'nogo zachatka.

Tissue engineering offers to restore the lost tooth using a biological analogue grown from the tooth germ. These technologies provide long-term cultivation of the germ in bioreactor in vitro. The subsequent transfer and growth of the in vitro grown tooth in the jaw is hampered by difficulty of integration of the new tooth with the host tissue. We suggested that growing tooth by homotopic transplantation in situ, that is, immediately in the jaw passing the in vitro stage will help to solve these problems. The aim of the work was to test the hypothesis. The principal possibility of transfer of the tooth germ directly into the jaw and cultivation in situ eliminating the stage in vitro is shown. The results showed a good integration of the grown teeth with the jaw without signs of inflammation and with the appearance of blood vessels in the pulp. At the same time, the results also showed the necessity to improve the preparation of tooth germs for transplantation and surgical procedures.

Cellular and molecular mechanisms of inflammation of esophageal mucosa under different.

The review presents modern data on the cellular and molecular mechanisms of inflammatory changes of esophageal mucosa exposed to different types of reluctate (gastric, biliary or duodenal/mixed). The authors describe data on key mediators of inflammation in gastroesophageal reflux disease (GERD) and their major cellular sources, changes of the immune profile of patients. Discusses the possible impact of changes in the cellular and molecular components in the development of the inflammatory response in the esophagus on the clinical features of GERD and its therapy-refractory forms.

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.

Effects of Phenotype of Retinal Macrophages on the Features of Angiogenesis of Murine Retina.

The period of forming of superficial vascular plexus during physiological retinal angiogenesis was shorter in C57Bl/6 mice. Experiments on the model of oxygen-induced retinopathy showed that avascular and vascularized zones in BALB/c mice on day 17 are smaller than in C57Bl/6 mice are by 5 and 1.5 times, respectively. The obtained results confirmed the importance of phenotype of retinal macrophages in the regulation of processes of both physiological and pathological retinal angiogenesis.

[An immune response and an alveolar macrophage phenotype in asthma, gastroesophageal reflux disease and their concurrence].

AIM: To test the hypothesis that an impaired pulmonary immune response in asthma, gastroesophageal reflux disease (GERD) and their concurrence of these diseases is largely determined by disordered alveolar macrophage (AM) reprogramming and to assess the pulmonary immune response and an AM phenotype in patients with asthma, GERD and their concurrence. SUBJECTS AND METHODS: The levels of proinflammatory M1 cytokines, such as IL-1ß, IL-8, IL-12p70, IFN-γ, TNF-α, and TNF-ß, anti-inflammatory M2 cytokines, such as IL-4, IL-5, and IL-10, and bivalent M1/M2 cytokines, such as IL-2 and IL-6, were determined in bronchoalveolar lavage fluid (BALF) and AM culture medium. RESULTS: Serious deformations in the pulmonary immune response were first detected in patients with mixed pathology towards to an anti-inflammatory M2 phenotype. The change in the pulmonary immune response phenotype in GERD towards Ml and in comorbidity towards M2 was coincident with that of the AM phenotype. In asthma, the change in the pulmonary immune response phenotype occurred towards to M2 and that in the intrinsic AM phenotype did towards M1. This phenotype is likely to form a proinflammatory component and to cause an asthma exacerbation. CONCLUSION: Analysis of the spectrum of cytokines in BALF and produced by macrophages in asthma, GERD and their concurrence validated the hypothesis that impaired pulmonary immune responses in these diseases are associated with disordered AM reprogramming. The findings also suggest that therapy for the inflammatory component in these diseases should be performed by taking into account the specificity of the cytokine structure of an immune response and the phenotypic heterogeneity of immune cells.

[Phenomena and signaling mechanisms of macrophage reprogramming].

The process of changing the phenotype of macrophages is called <>. Reprogramming plays a central role in the immune response. Reprogrammed macrophages are characterized by four phenomena: the phenomenon of the gain response of macrophages on the reprogramming factor (direct gain), and the other factor (cross-gain); phenomenon of reciprocal suppression of alternative phenotypes; phenomenon of cascade activation for mechanisms of reprogramming and the phenomenon of positive and negative feedbacks. The formation of these phenomena provide the intracellular signaling pathways, such as INK-; PI3K/Akt-; Notch-; JAK/STAT-; TGF-ß-; TLR/NF-κB- and hypoxia-dependent signaling pathways. Analysis of the signaling mechanisms of reprogramming led to several conclusions: 1. There is a relative specialization of signaling pathways in macrophages reprogramming on action of different components of the microenvironment; 2. signaling pathways that are involved in reprogramming of macrophages can be divided in the way that program M1 phenotype and the way that program M2 phenotype; 3. Understanding the signaling pathways helps to explain the basic phenomena of reprogramming. Thus, the phenomenon of the gain response of reprogrammed macrophages is provided by convergence signaling pathways at specific protein; phenomenon of reciprocal suppression alternative macrophage phenotype is provided by that the formation of one phenotype is accompanied by increased synthesis of molecules that inhibit an alternative phenotype; at the heart of the phenomenon of cascade activation of signaling pathways is the ability one way to transmit signals over a different path and the basis for the phenomenon of positive and negative feedback is the ability to increase the synthesis of the activators and inhibitors of this pathway. 4. Signalling pathways that transmit the signal from the proinflammatory factors and programm proinflammatory M1 phenotype of macrophages are often branching, which, when activated, may increase the production of anti-inflammatory M2 cytokines; and vice versa. Since the violation reprogramming of macrophages plays an important role in the development of many diseases, understanding the signaling mechanisms of reprogramming, will assist in the selection of effective therapeutic targets to develop new ways of correction of impaired immunity.

[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.

[Effect of adaptation to hypoxia on expression of NO synthase isoforms in rat myocardium].

Previously we have shown that adaptation to hypoxia (AH) is cardio- and vasoprotective in myocardial ischemic and reperfusion injury and this protection is associated with restriction of nitrosative stress. The present study was focused on further elucidation of NO-dependent mechanisms of AH by identifying specific NO synthases (NOS) that could play the major role in AH protection. AH was performed in a normobaric hypoxic chamber by breathing hypoxic gas mixture (9.5-10% O2) for 5-10 min with intervening 4 min normoxia (5-8 cycles daily for 21 days). Expression of neuronal (nNOS), inducible (iNOS), and endothelial (eNOS) protein was measured in the left ventricular myocardium using Western blot analysis with respective antibodies. AH educed iNOS protein expression by 71% (p < 0.05) whereas eNOS protein expression tended to be reduced by 41% compared to control (p < 0.05). nNOS protein expression remained unchanged after AH. Selective iNOS inhibition can mimic the AH-induced protection. Therefore protective effects of AH could be at least partially due to restriction of iNOS and, probably, eNOS expression.

[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.

Alternative reprogramming of M1/M2 phenotype of mouse peritoneal macrophages in vitro with interferon-γ and interleukin-4.

An important role in the development of the immune response is played by macrophages that acquire either anti-inflammatory M1 or anti-inflammatory M2 phenotype depending on their microenvironment. The possibility of targeted reprogramming of the initial M2 macrophage phenotype towards M1 phenotype and vice versa using macrophage reprogramming factors IFN-γ and IL-4, respectively, was demonstrated. We showed that macrophages of genetically different mouse strains did not practically differ by their reprogramming capacity. Our findings suggest that macrophage programming not only participates in the triggering of the immune response, but also can ensure plasticity of functional activity during the developing response.

SP-D-dependent regulation of NO metabolism in lipopolysaccharide-stimulated peritoneal macrophages.

This work was designed to study the role of surfactant protein D in the regulation of NO synthesis by "non-alveolar" microphages. We evaluated whether the effects of surfactant protein D depend on the phenotype of macrophages. In the absence of surfactant protein D, the LPS-induced iNOS response was shown to decrease in macrophages of native and proinflammatory phenotypes by 30%, and in macrophages of the antiinflammatory phenotype (by 63%). Under the influence of lipopolysaccharide in high doses (500 ng/ml), NO(2)*- production by mouse macrophages without surfactant protein D was reduced in native cells (by 25%), but increased in proinflammatory (by 40%) and antiinflammatory phenotypes (by 12% compared to mouse macrophages with surfactant protein D). Our results suggest that surfactant protein D is involved in the immune response in the whole organism, but not only in the lungs. The effect of surfactant protein D depends on the phenotype of macrophages.

Controlled modulation of inflammatory, stress and apoptotic responses in macrophages.

An outstanding question of current immunology is to define the mechanisms by which microbial products influence the immunopathologic host response elements in the early stages of infection. Macrophages are now well recognized to have a critical role in both innate and acquired immunity. In order to adjust promptly to continuous changes in microenvironment and maintain the immunologic balance, macrophages adequately respond by activating one of the numerous immunologic programs. However, sustained macrophage activation and excessive production of inflammatory mediators can perpetuate the numerous pathological processes and contribute to induction of stress response and even apoptosis. Therefore, selective modulation of macrophage activity represents an important strategy for prevention and treatment of inappropriate inflammatory responses in order to minimize the unwanted side-effects of the immunity. Macrophages can be selectively reprogrammed for a specific phenotype of immune response, e.g. cytokine or nitric oxide (NO), by relatively short-term exposure of the cells to substimulatory concentrations of different microbial components, including LPS. These LPS-dependent reprogramming effects are mediated by IFN-gamma-independent autocrine cytokine regulatory mechanisms that also controlled at the transcriptional level. Furthermore, LPS reprogrammed macrophages exhibit differential capacity to resist experimentally induced apoptosis and to produce heat shock proteins. Complete analysis of, and appreciation for, the immunoregulatory mechanisms implicated in LPS-dependent reprogramming of immune responses in macrophages can be expected to increase our understanding of the host innate response, as well as allow investigators to utilize emerging immunologic technologies in effective treatment of infections and chronic inflammatory diseases.

Prevention of neurodegenerative damage to the brain in rats in experimental Alzheimer's disease by adaptation to hypoxia.

We report here studies addressing the possibility of preventing neurodegenerative changes in the brain using adaptation to periodic hypoxia in rats with experimental Alzheimer's disease induced by administration of the neurotoxic peptide fragment of beta-amyloid (Ab) into the basal magnocellular nucleus. Adaptation to periodic hypoxia was performed in a barochamber (4000 m, 4 h per day, 14 days). The following results were obtained 15 days after administration of Ab. 1. Adaptation to periodic hypoxia significantly blocked Ab-induced memory degradation in rats, as assessed by testing a conditioned passive avoidance reflex. 2. Adaptation to periodic hypoxia significantly restricted increases in oxidative stress, measured spectrophotometrically in the hippocampus in terms of the content of thiobarbituric acid-reactive secondary lipid peroxidation products. 3. Adaptation to periodic hypoxia completely prevented the overproduction of NO in the brains of rats with experimental Alzheimer's disease, as measured in terms of increases in tissue levels of stable NO metabolites, i.e., nitrites and nitrates. 4. The cerebral cortex of rats given Ab injections after adaptation to periodic hypoxia did not contain neurons with pathomorphological changes or dead neurons (Nissl staining), which were typical in animals with experimental Alzheimer's disease. Thus, adaptation to periodic hypoxia effectively prevented oxidative and nitrosative stress, protecting against neurodegenerative changes and protecting cognitive functions in experimental Alzheimer's disease.

Specific response of the organism and blood leukocytes in rats of different genetic strains to hypoxia.

By the open-field behavior, August rats were more resistant to acute hypoxia than Wistar rats. Hypoxic activation of the immune system was more pronounced in August rats. As differentiated from Wistar rats, the stress-limiting NO system in August rats was not suppressed during hypoxia. The effectiveness and resistance of this system to hypoxia were higher in August rats. Behavioral changes in Wistar rats under hypoxic conditions were accompanied by activation of HSP32 synthesis in blood leukocytes. This protein serves as an indicator of oxidative stress (i.e. adverse factor in hypoxia). August rats were more resistant to behavioral disturbances in hypoxia than Wistar rats. HSP32 synthesis in leukocytes from August rats was not impaired under hypoxic conditions. Our results indicate that variations in HSP32 synthesis in peripheral blood leukocytes can be considered as a matter of for resistance to acute hypoxia.

Role of nitric oxide in prevention of cognitive disorders in neurodegenerative brain injuries in rats.

NO synthesis disturbances play an important role in the development of neurodegenerative damage in Alzheimer disease. We previously showed that adaptation to intermittent hypobaric hypoxia prevents cognitive disturbances in rats with experimental Alzheimer disease. Here we evaluated the role of NO in cognitive disorders and development of adaptive protection during experimental Alzheimer disease. Adaptation to hypoxia in rats was performed in a hypobaric pressure chamber at a simulated altitude of 4000 m (4 h per day for 14 days). Alzheimer disease was simulated by bilateral injections of a toxic fragment of beta-amyloid (25-35) into n. basalis magnocellularis. For evaluation of the role of NO in the development and prevention of memory disorders, the rats received intraperitoneally either NO-synthase inhibitor N omega-nitro-L-arginin (L-NNA, 20 mg/kg, every other day for 14 days) or NO-donor dinitrosyl iron complex (200 microg/kg daily for 14 days). NO-synthase inhibitor potentiated the damaging effect of beta-amyloid, abolished the protective effect of adaptation to hypoxia, and produced memory disorders in rats similar to those observed during experimental Alzheimer disease. In contrast, the increase in NO level in the body provided by injections of the NO-donor produced a protective effect against memory disorders caused by beta-amyloid similar to that induced by adaptation to hypoxia. We concluded that reduced NO production in the organism plays an important role in the development of cognitive disorders produced by injections of beta-amyloid, while prevention of NO deficit by administration of NO-donors or non-pharmacological stimulation of NO synthesis can provide a protective effect in experimental Alzheimer disease.

Intracellular and extracellular NO differentially modulates the stress response and apoptosis in macrophages exposed to the influence of biological or physical factors.

We studied the role of extracellular and intracellular NO in the regulation of the stress response and apoptosis in macrophages of proinflammatory and antiinflammatory phenotypes under the influence of S. aureus and heat shock. Blockade of extracellular nitric oxide synthesis in cells with antiinflammatory phenotype inhibited the stress response induced by S. aureus and heat shock. The decrease in extracellular nitric oxide concentration around antiinflammatory macrophages potentiated the stress response induced by S. aureus, but had no effect on the stress response induced by heat shock. Hence, intracellular NO mediates the stress response induced by S. aureus and heat shock, while extracellular NO inhibits the stress response induced by S. aureus, but has no effect on the stress response induced by heat shock. In cells with antiinflammatory phenotype, intracellular NO plays an antiapoptotic role. S. aureus and heat shock did not cause apoptosis in macrophages with proinflammatory phenotype, while intracellular NO did not play a role in antiapoptotic activity of the proinflammatory phenotype. Extracellular NO synthesized by macrophages protects these cells from apoptosis induced by S. aureus and heat shock.

Antiarrhythmic effect of heat adaptation in ischemic and reperfusion injury to the heart.

Study on a model of 6-day dosed adaptation to heat in rats showed that this adaptation decreased the severity of cardiac arrhythmias during ischemic and reperfusion injury. The duration of arrhythmias decreased not only in the ischemic period, but also under conditions of reperfusion. Adaptation delayed the development of arrhythmias during ischemia, decreased the number of animals with late reperfusion arrhythmias, and improved recovery of the heart after ischemia and reperfusion.