To date, much remains unclear about the pathogenesis of asthma, one of the most common chronic and highly heterogenic diseases of the respiratory system. The lack of specific and highly effective therapy in case of certain asthma subtypes requires the search for new approaches to treatment. One possible approach would be to influence the metabolism and immune functions of myeloid cells. This approach finds its application in the treatment of cancer and other diseases in the pathogenesis of which macrophages play an important role. It was shown that the pathogenesis of allergic asthma in response to one of the most common allergens, house dust mite, is due to a metabolic TNF-mediated reprogramming of alveolar macrophages. This suggests that influencing the process of TNF production or metabolic adaptations with specific blockers may also lead to a reduction in the symptoms of the course of the disease as a whole. In this work, we experimentally tested whether the previously obtained phenotype that occurs in macrophages in response to HDM cultured in DMEM is preserved if cells are cultured under more physiologically relevant conditions: in a medium closely related in composition to blood plasma. We also analyzed open databases of alveolar macrophages sequencing obtained from patients with asthma or from the lungs of mice in an HDM-induced asthma model in order to correlate specific immunometabolic changes. It was found that macrophages cultured under conditions close to physiological, simultaneously increase the rates of respiration and glycolysis, and also produce TNF in response to HDM. The observed phenotype is consistent with transcriptomic analyzes performed on human and mouse samples, which revealed an increase in the expression of genes related to glycolysis, oxidative phosphorylation, and the TNF signaling pathway. Thus, the data confirm the relevance of the phenotype obtained in vitro to the changes occurring in the in vivo system. However, functional verification at the level of metabolites, proteins and changes in metabolic activity is also required. In addition, it remains to be established how the blocking of individual metabolic pathways affects the features of the functional macrophage phenotype that occurs in response to HDM, and whether this effect can alleviate asthma symptoms.

Itaconate is an immunoregulatory metabolite produced by myeloid cells and plays a key role in the regulation of the immune response. Itaconate, on the one hand, is able to suppress the activity of succinate dehydrogenase (SDH), thereby making a significant contribution to the metabolic reprogramming of the cell. On the other hand, itaconate can regulate the activity of a number of transcription factors and transcription regulators, thereby affecting gene expression. In most experimental studies, itaconate has been characterized predominantly as an anti-inflammatory agent. In particular, itaconate produced by activated macrophages inhibits the production of cytokines TNF, IL-1b, IL-6, IL-10. However, some evidence suggests a pro- inflammatory role for itaconate in a number of mouse disease models. Thus, the deletion of the Acod1 gene responsible for the production of itaconate leads to the suppression of the production of TNF and IL-6 in the mouse polymicrobial sepsis model, which means that in the context of inflammation in vivo , itaconate can act as an inducer of pro-inflammatory cytokines. The mechanism of itaconate regulation of cytokine production in systemic inflammation remains unexplored. In this work, we have shown that injection of itaconate and its derivative dimethyl itaconate into mice, followed by induction of inflammation by bacterial lipopolysaccharide (LPS), leads to changes in the content of cytokines in the blood. Interestingly, the systemic production of IL-6 and IL-10 in response to itaconate is increased, contrary to the results previously obtained in cell cultures. At the same time, IFNg production, on the contrary, is suppressed. Apparently, itaconate regulates the production of cytokines in vivo by suppressing the activity of SDH. Injection of the SDH inhibitor, dimethylmalonate, followed by induction of inflammation in mice, results in similar changes in blood cytokines observed in response to itaconate: increased production of IL-6, IL-10 and suppression of IFNg production. On the contrary, the addition of succinate, a SDH substrate, leads to the opposite effect on cytokine production. Thus, it can be assumed that the observed effects of itaconate on cytokine production in the model of LPS-induced inflammation are mediated by its ability to inhibit SDH. These results help to understand the controversial role of itaconate in inflammation and shed light on a previously undescribed relationship between SDH and cytokine production in inflammation in vivo .

Systemic cytokine inhibition may be an effective therapeutic strategy for several autoimmune diseases. However, recent studies suggest that tissue or cell type-specific targeting of certain cytokines, including TNF, may have distinct advantages and show fewer side effects. Here we describe protocols for generating and testing bispecific cytokine inhibitors using variable domain of single-chain antibodies from Camelidae (VHH) with a focus on cell-specific TNF inhibitors.

Mechanisms of skin wound healing have been extensively studied in order to propose novel approaches for treatment of chronic and poorly healing wounds, including those associated with autoimmune pathologies, as well as for fibrosis prevention. Recent studies indicate the key role of proinfl ammatory cytokines, such as TNF, IL-6 and IL-1, in regulating skin regeneration. In our work we investigated the impact of tumor necrosis factor (TNF) on healing of deep skin wounds in mice by employing reverse genetics approach. We found, that conditional knock-out of TNF gene specifically in macrophages results in delayed wound healing. However inactivation of TNF in all cell types does not aff ect wound regeneration dynamics. Moreover, genetic knock-out of TNF receptor I (TNFRI) results in accelerated wound healing. Thus, the results of our study suggest that TNF from diff erent cell types plays a dual role in skin regeneration probably refl ecting that both cellular source of TNF as well as receptor type on target cells are important.

Biomaterials have recently been recognized for their potential use in regenerative medicine and tissue engineering, since they not only represent a scaff old for cell populations, but also can infl uence functionality of diff erent cell types. In particular silk-based biomaterials (i. e. fibroin or spidroin) are of high interest, because they show prominent mechanistic features and have proregenerative potential. In our work we showed that composite fibroin/gelatin scaff olds induce moderate infl ammation both in vitro and in vivo and this eff ect depends on the configuration of the matrices. In particular, culturing of Mouse Embryonic Fibroblasts (MEF) on three-dimensional (3D), but not two-dimensional (2D) fibroin/gelatin scaff olds resulted in overexpression of adhesion molecules ICAM-1 and VCAM-1 as well as proinfl ammatory cytokines TNF and IL-6. Moreover subcutaneous injection of fibroin/gelatin microparticles in mice lead to accumulation of infl ammatory myeloid cells and acceleration of skin wound healing, while implantation of 3D fibroin/gelatin scaff olds under the kidney capsule promoted lymphocytes clusterization, resulting in generation of primitive lymphoid tissue. Altogether our data suggest immunomodulatory features of fibroin/gelatin scaff olds, which can be useful in tissue engineering and regenerative medicine.

Cytokines comprise the molecular language of communication between the cells, which is needed to maintain the homeostatic functions of the body (including the immune system) and mediate various diseases. Many aspects of inflammation, autoimmune diseases and neoplasia are associated with cytokine signaling through specific receptors. The establishment of new physiological functions of "old" cytokines and understanding the molecular and cellular mechanisms of their involvement in disease pathogenesis, as well as the search for new therapeutic targets and development of innovative approaches to anti-cytokine therapy, present a fundamental problem. When assessing the tremendous success of anti-cytokine therapy in treatment of certain autoimmune diseases, we should not forget that (a) this treatment does not eliminate the causes of the disease:autoreactive T-cell clones; and that (b) less than half of the patients respond to this therapy; and that (c) anti-cytokine therapy has serious side effects.

Skin wound healing is subject to an intricate regulation, involves many cell populations and molecular mediators, and is one of the key mechanisms that ensures the barrier functions of the skin and the maintenance of body homeostasis. The efficiency of this process is largely determined by the balance of proinflammatory and proregenerative signals, which are mediated by cytokines. The review summarizes the latest data on the role of proinflammatory cytokines, mainly tumor necrosis factor (TNF), interleukin 6 (IL-6), interleukin 1 (IL-1), and interferons (IFNs), in skin wound healing, including those obtained with the use of genome editing techniques and methods of reverse genetics to establish relevant animal models. The roles that proinflammatory cytokines play at various stages of skin regeneration are discussed for both normal state and systemic pathologies, such as diabetes. Promising approaches to treating poorly healing wounds are summarized.

To model human interleukin-6 (hIL-6) associated diseases, unique mice with transgenic overexpression of human IL-6 and reporter fluorescent protein EGFP in cells of macrophage-monocyte lineage were generated using loxP–Cre system. High level of hIL-6 production by macrophages and monocytes, as confirmed in vitro in primary culture of bone marrow-derived macrophages, in vivo resulted in early postnatal death in vivo, presumably, due to the effect of overexpression of hIL-6 on hematopoiesis.

Reverse genetics approach, involving genome editing, makes it possible not only to establish the nonredundant and unique functions of genes and their products, but also to construct animal models for biomedical research. Interleukin 6 (IL-6) is an important immunoregulatory and proinflammatory cytokine that differs from many related proteins in having a rather complicated signal transduction scheme. Apart from the multiple functions of IL-6, the most relevant biological problem of recent years was establishing what cells produce IL-6, in what form IL-6 is produced, what cells are recipients of the IL-6 signal, and what are the downstream events and physiological consequences of the IL-6 signaling cascade. Because IL-6 is involved in the pathogenesis of many diseases and is a drug target, understanding the mechanisms of its normal and pathogenic effects is important for the clinics. The review summarizes the recent data available in the field.

Reverse genetics approach, involving genome editing, makes it possible not only to establish the nonredundant and unique functions of genes and their products, but also to construct animal models for biomedical research. Interleukin 6 (IL-6) is an important immunoregulatory and proinflammatory cytokine that differs from many related proteins in having a rather complicated signal transduction scheme. Apart from the multiple functions of IL-6, the most relevant biological problem of recent years was establishing what cells produce IL-6, in what form IL-6 is produced, what cells are recipients of the IL-6 signal, and what are the downstream events and physiological consequences of the IL-6 signaling cascade. Because IL-6 is involved in the pathogenesis of many diseases and is a drug target, understanding the mechanisms of its normal and pathogenic effects is important for the clinics. The review summarizes the recent data available in the field.

Culturing of allogeneic or autologous cells in three-dimensional bioresorbable scaffolds is an important step in the engineering of constructs for regenerative medicine, as well as for experimental systems to study the mechanisms of cell differentiation and cell-to-cell interaction. Artificial substrates can modulate the phenotype and functional activity of immobilized cells. Investigating these changes is important for understanding the fundamental processes underlying cellular interactions in a 3D microenvironment and for improving tissue-engineered structures. In this study, we investigated the expression of the ICAM-1 adhesion molecule in mouse embryonic fibroblasts (MEF) when cultured on gelatin-fibroin scaffolds. Increased expression of ICAM-1 in MEF was detected only under 3D culture conditions both at the mRNA and protein levels. At the same time, the MEF cultured on various substrates did not oerexpress MAdCAM-1, indicating the selective effect of 3D culture conditions on ICAM-1 expression. One possible mechanism for ICAM-1 induction in MEF is associated with the activation of AP-1, since expression of c-Fos and Junb (but not cJun and Jund) was increased in MEF in 3D. When cultured under 2D conditions, the expression level of AP-1 components did not change.

This review addresses the issue of bioengineering of artificial lymphoid organs. Progress in this field may help to better understand the nature of the structure-function relations that exist in immune organs. Artifical lymphoid organs may also be advantageous in the therapy or correction of immunodefficiencies, autoimmune diseases, and cancer. The structural organization, development, and function of lymphoid tissue are analyzed with a focus on the role of intercellular contacts and on the cytokine signaling pathways regulating these processes. We describe various polymeric materials, as scaffolds, for artificial tissue engineering. Finally, published studies in which artificial lymphoid organs were generated are reviewed and possible future directions in the field are discussed.

The process of tissue regeneration following damage takes place with direct participation of the immune system. The use of biomaterials as scaffolds to facilitate healing of skin wounds is a new and interesting area of regenerative medicine and biomedical research. In many ways, the regenerative potential of biological material is related to its ability to modulate the inflammatory response. At the same time, all foreign materials, once implanted into a living tissue, to varying degree cause an immune reaction. The modern approach to the development of bioengineered structures for applications in regenerative medicine should be directed toward using the properties of the inflammatory response that improve healing, but do not lead to negative chronic manifestations. In this work, we studied the effect of microcarriers comprised of either fibroin or fibroin supplemented with gelatin on the dynamics of the healing, as well as inflammation, during regeneration of deep skin wounds in mice. We found that subcutaneous administration of microcarriers to the wound area resulted in uniform contraction of the wounds in mice in our experimental model, and microcarrier particles induced the infiltration of immune cells. This was associated with increased expression of proinflammatory cytokines TNF, IL-6, IL-1β, and chemokines CXCL1 and CXCL2, which contributed to full functional recovery of the injured area and the absence of fibrosis as compared to the control group.

Interleukin-6 (IL-6)--one of the most important pro-inflammatory cytokines that has a broad spectrum of immunoregulatory properties. Molecular mechanisms of signal transduction of IL-6 and its receptor, which were previously established, have recently been supplemented with a concept of trans-signaling. Selective inhibition of this signaling cascade would allow to modulate the pathological effects of IL-6. Methods of reverse genetics have helped to establish the physiological functions of IL-6 in normal state and in various diseases, including neoplasias. Therapeutic inhibitors of IL-6 or its receptor are already used for the treatment of several autoimmune diseases, however, systemic inhibition inevitably also neutralizes the protective functions of this cytokine. It is expected that in the future systemic therapy will be replaced by more specific and effective approaches that take into account the peculiarities of molecular signaling pathways in target cells and differences in the function of IL-6, depending on the cell source.

Обзор посвящен проблеме конструирования искусственных лимфоидных органов. Успех в этой области позволит не только лучше понять механизмы функционирования нормальных органов иммунной системы, но и разработать новые подходы к терапии иммунодефицитов, аутоиммунных состояний и, возможно, других заболеваний. На примере лимфатического узла мыши рассмотрено строение и развитие нормальных лимфоидных органов. Особое внимание уделено роли межклеточных взаимодействий и цитокиновых сигналов в механизмах формирования и функционирования лимфоидных органов. Описаны биоматериалы, на основе которых возможно создание искусственных органов, в том числе органов иммунной системы. Критически рассмотрены достижения последних лет в области биоинженерии искусственных лимфоидных органов и предполагаемые направления исследований.

Interleukin-6 (IL-6) is one of the most important proinflammatory cytokines that has a broad spectrum of immunoregulatory properties. The molecular mechanisms of signal transduction of IL-6 and its receptor, which have been previously established, were later supplemented with a concept of trans-signaling. The selective inhibition of this signaling cascade would allow the modulation of the pathological effects of IL-6. Reverse genetics methods helped to establish the physiological functions of IL-6 in normal state and in various diseases, including neoplasias. Therapeutic inhibitors of IL-6 or its receptor are already used to treat several autoimmune diseases; however, the systemic inhibition inevitably also neutralizes the protective functions of this cytokine. It is expected that, in the future, systemic therapy will be replaced by more specific and effective approaches that take into account the peculiarities of molecular signaling pathways in target cells and differences in the function of IL-6 depending on the cell source.

Experimental a way the close numeral values of linear active electric resistance and specific conductivity of heavy-current plasma channel of an air heavy-current of discharge are got in the chain of powerful capacity store of energy, in-use at forming repeated impulsive components of current of artificial lightning with the rationed amplitude to 100 k A.