The development of CAR-T specific therapy made a revolution in modern oncology. Despite the pronounced therapeutic effects, this novel approach displayed several crucial limitations caused by the complications in pharmacokinetics and pharmacodynamics controls. The presence of the several severe medical complications of CAR-T therapy initiated a set of attempts aimed to regulate their activity in vivo. We propose to apply the barnase-barstar system to control the cytotoxic antitumor activity of CAR-T cells. To menage the regulation targeting effect of the system we propose to use barstar-modified CAR-T cells together with barnase-based molecules. Barnase was fused with designed ankyrin repeat proteins (DARPins) specific to tumor antigens HER2 (human epidermal growth factor receptor 2) The application of the system demonstrates the pronounced regulatory effects of CAR-T targeting.

We previously showed that the metal-binding domain 1-16 of intact amyloid-beta (Aβ) is involved in interactions with a number of proteins from the cytosolic fraction of SK-N-SH human neuroblastoma cells in a zinc-dependent manner only. It is known that hereditary mutations in the Aβ metal-binding domain (Aβ(1-16)), which accelerate the development of Alzheimer's disease and post-translational modifications of amino acid residues, can significantly affect the domain's structure in the presence of zinc ions. In this work, using the molecular fishing methodology for Aβ(l-16) isoforms with the Taiwanese mutation (D7H) and a phosphorylated Ser8 residue, proteins from the cytosol of SK-N-SH cells were found that are able to form zinc-dependent non-covalent complexes with these domains. The partner proteins identified for these isoforms differed from those for intact Aβ(1-16). In contrast, the Aβ(1-16) isoform with the English mutation (H6R) and the Aβ(1-16) isoform containing both an isomerized Asp7 residue and phosphorylated Ser8 residue did not interact with cytosolic proteins. The results are useful for developing methods for rational modulation of protein-protein interactions involving natural isoforms of beta-amyloid, and also indicate the possible role of beta-amyloid with phosphorylated Ser8 as a molecule involved in normal physiological processes.

By now, a whole number of pathogenic antibiotic-resistant or tolerant microorganisms has been progressively increased. Hence, efficient fight against them requires to change the class of antibiotics, increase their dose, or develop new antimicrobial drugs. On the contrary, another option could rely on augmenting innate immunity. During coevolution, eukaryotes have developed several ways for their protection against microorganisms. Innate immunity conserved in all multicellular organisms. The essential principles of innate immunity include recognition of a foreign structures and their subsequent destruction. A set of specific receptors recognize conserved pathogen-derived structures. Elimination occurs due to phagocytosis and cleavage, e.g. via oxidative burst in phagocytic cells, compliment system or antimicrobial peptides. Recognition system in innate immunity is based on the pattern recognition receptors. Due to the pathogen diversity, multiple conserved structures typical to pathogens (e.g. lipopolysaccharide, peptidoglycan, flagellin etc.) are sensed by numerous receptors. The family of peptidoglycan recognition proteins is among such receptors, which were first isolated in 1996 from the silkworm Bombyx mori and mice. Later, it was demonstrated that this family is conserved and its members are found in insects, fish and mammals. Here, functions of insect peptidoglycan recognition proteins in Drosophila melanogaster as well as mammals are discussed. Such proteins are expressed mainly in liver cells (insects — in adipose tissue cells as analogue of mammalian liver), intestinal cells, and epidermis. Numerous studies demonstrate that peptidoglycan-recognition proteins moderate immune response, and may act as antimicrobial proteins, or to regulate microbiota as well as prevent enterocyte activation and restrict inflammatory response. Due to evolutionary conservatism observed for such proteins and inability for bacteria to evade their protective effects, it seems promising to use peptidoglycan recognition proteins in a combination therapeutic approach against antibiotic-resistant and antibiotic-tolerant forms of microorganisms.

Tag7 (PGRP-S) is an innate immune protein that is involved in the antibacterial and antitumor defense and stimulates the maturation of cytotoxic lymphocyte subpopulations. It was found that the incubation of lymphocytes with Tag7 for 3 days promotes the appearance of cytotoxic NK cells that are active against a number of tumor cell lines.

Periodontal diseases, especially those with polymicrobial etiology, are often associated with type 2 diabetes mellitus, proceeding more severely and affecting the course of diabetes mellitus. Recently, this feature has been associated with the ability of periodontopathogen microflora to cause not only a local infectious process in the oral cavity, but also to interact with the human immune system and induce various systemic effects. We investigated changes in the salivary cytokine profile of patients with chronic periodontitis, associated and not associated with type 2 diabetes mellitus. We observed a statistically significant decrease of MCP-1/CCL2, GM-CSF, IL-5, IL-6, and IFN- in the saliva of patients with chronic periodontitis associated with type 2 diabetes mellitus in comparison with patients with chronic periodontitis only. All of these cytokines are associated with macrophage activation. These data are an important contribution to the elucidation of the mechanism of periodontopathogens involvement in the manifestation of the systemic effects of type 2 diabetes.

Tag7 (PGRP-S) – белок врождённого иммунитета, который принимает участие в антибактериальной и противоопухолевой защите, а также стимулирует созревание цитотоксических субпопуляций лимфоцитов. Обнаружили, что инкубация в течение 3 сут лимфоцитов с Tag7 способствует появлению цитотоксических NK-клеток, активных против ряда опухолевых клеточных линий.

Glycolysis activation is one of the main features of energy metabolism in cancer cells that is associated with the increase in glycolytic enzyme synthesis, primarily, hexokinases (HKs), in many types of tumors. Conversely, in colorectal cancer (CRC) the decrease in the expression of HK2 gene, which encodes one of the key rate-limiting enzyme of glycolysis, was revealed, thus, the study of the mechanisms of its inhibition in CRC is of particular interest. To search for potential microRNAs, inhibiting the expression of HK2 in CRC, we have performed the analysis of data from "The Cancer Genome Atlas" (TCGA) and five microRNA-mRNA target interaction databases (TargetScan, DIANA microT, mirSVR (miRanda), PicTar, and miRTarBase) using original CrossHub software. Seven microRNAs containing binding site on mRNA HK2, which expression is negatively correlated with HK2 expression, were selected for further analysis. The expression levels of these microRNAs and mRNA HK2 were estimated by quantitative PCR on a set of CRC samples. It has been shown, that the expression of three microRNAs (miR-9-5p, -98-5p, and -199-5p) was increased and correlated negatively with mRNA level of HK2 gene. Thus, downregulation of HK2 gene may be caused by its negative regulation through microRNAs miR-9-5p, -98-5p, and -199-5p.

Glycolysis activation is one of the main features of energy metabolism in cancer cells that is associated with the increase in glycolytic enzyme synthesis, primarily, hexokinases (HKs), in many types of tumors. Conversely, in colorectal cancer (CRC) the decrease in the expression of HK2 gene, which encodes one of the key rate-limiting enzyme of glycolysis, was revealed, thus, the study of the mechanisms of its inhibition in CRC is of particular interest. To search for potential microRNAs, inhibiting the expression of HK2 in CRC, we have performed the analysis of data from "The Cancer Genome Atlas" (TCGA) and five microRNA-mRNA target interaction databases (TargetScan, DIANA microT, mirSVR (miRanda), PicTar, and miRTarBase) using original CrossHub software. Seven microRNAs containing binding site on mRNA HK2, which expression is negatively correlated with HK2 expression, were selected for further analysis. The expression levels of these microRNAs and mRNA HK2 were estimated by quantitative PCR on a set of CRC samples. It has been shown, that the expression of three microRNAs (miR-9-5p, -98-5p, and -199-5p) was increased and correlated negatively with mRNA level of HK2 gene. Thus, downregulation of HK2 gene may be caused by its negative regulation through microRNAs miR-9-5p, -98-5p, and -199-5p.

Naïve non-activated lymphocytes are capable of releasing the chemoattractant complex Tag7–Mts1 and can migrate along the gradient of its concentration. After activation of these cells by IL-2, they acquire the abilities to kill tumor cells and to release the cytotoxic Tag7–Hsp70 complex, which is accompanied by a loss of both the Tag7–Mts1-mediated lymphocyte chemotaxis and the ability to release this chemoattractant into the conditioned medium.

FN-y is a cytokine of great clinical significance. The excessive or insufficient function of the signalling pathway triggered by IFN-y is a cause of various pathological states of the immune system and the body in general. At present, clinical practice uses quantitative methods of detection of this protein without taking into account its functional activity. Traditional tests for assessment of the biological activity of IFN-y are technologically complex and their results are poorly quantified. In the course of this work a model system was created representing a cell line with a reporter construct stably integrated into the genome controlled by a STAT-dependent promotor. Processing of the cells of this model system by IFN-y activates the STAT-1-signalling pathway, which causes expression of FFly luciferase gene. Luciferase activity is quantitatively assessed by detection of bioluminescence. Working concentration values (2.5 ng/ml) and time of performing the assay (8 h) were chosen. The effectiveness of using the model system for testing IFN-y bioactivity has been shown.

FN-y is a cytokine of great clinical significance. The excessive or insufficient function of the signalling pathway triggered by IFN-y is a cause of various pathological states of the immune system and the body in general. At present, clinical practice uses quantitative methods of detection of this protein without taking into account its functional activity. Traditional tests for assessment of the biological activity of IFN-y are technologically complex and their results are poorly quantified. In the course of this work a model system was created representing a cell line with a reporter construct stably integrated into the genome controlled by a STAT-dependent promotor. Processing of the cells of this model system by IFN-y activates the STAT-1-signalling pathway, which causes expression of FFly luciferase gene. Luciferase activity is quantitatively assessed by detection of bioluminescence. Working concentration values (2.5 ng/ml) and time of performing the assay (8 h) were chosen. The effectiveness of using the model system for testing IFN-y bioactivity has been shown.

The results of comparative analysis of interaction between the protein cytotoxic complex Tag 7–Hsp70 and the Tag 7 component of this complex with TNFR1 receptor in solution and in tumor cells are presented.

Treatment and diagnosis of acute lymphoblastic leukemias (ALL) in children have achieved much progress in the past years. Highly effective protocols for treatment of ALL have been developed, permitting to obtain remission in more than 90% of patients. Study of the level of minimal residual disease (MRD) permits to refer a patient to a certain group of risk, requiring a particular scheme of therapy and also to choose the optimal terms for bone marrow transplantation (BMT). Bone marrow (BM) samples of MRD-positive patients are very important for clarification of the mechanisms of tumour resistance to therapy. Introduction of new generation methods (high throughput sequencing, proteomics, bioinformatics) will permit to determine additional genetic and protein markers associated with higher levels of MRD, which in its turn might lead to creation of new effective markers and target therapeutic preparations.

An analysis of conformationally stable (conformationally conservative) tetrapeptides selected from protein structures that are deposited in the PDBSelect database has been carried out. The set contained 943 different tetrapeptide sequences, with each sequence occurring at least five times in different fragments of protein structure. Analysis of conformations based on DSSP markup revealed that the conformation of most peptides (900 of 943 sequences) was α-helical, while 43 sequences had different conformations, of which the left-handed polyproline II helix is especially worth mentioning. The physicochemical properties of conformationally stable peptides from each subset were inferred from the average hydropathicity of the tetrapeptides. The results of the calculations revealed “neutral” hydropathicity of the conformationally stable oligopeptides. Notably, the distribution of hydropathicity values for the conformationally stable peptides was considerably narrower than those for the control sets of peptides. Thus, conformationally stable oligopeptides form a specific group of local protein structures with nearly uniform conformational and physicochemical properties. The theory of specific long-range interactions that was previously developed by the authors of the present study assumes such peptides to be well adapted for efficient mutual recognition of molecules.

PGRP-S (Tag7) is an innate immunity protein involved in the antimicrobial defense systems, both in insects and in mammals. We have previously shown that Tag7 specifically interacts with several proteins, including Hsp70 and the calcium binding protein S100A4 (Mts1), providing a number of novel cellular functions. Here we show that Tag7–Mts1 complex causes chemotactic migration of lymphocytes, with NK cells being a preferred target. Cells of either innate immunity (neutrophils and monocytes) or acquired immunity (CD4+ and CD8+ lymphocytes) can produce this complex, which confirms the close connection between components of the 2 branches of immune response.

The goal of our research was comparative study of the most important parameters of subset cytoarchitectonics in the patients with the different courses of myocarditis and evaluation of their pathoge$ netic and clinical value in the practice of the physician. We have investigated 99 patients with myocarditis and 40 healthy donors. In patients with malignant course of

Abstract. Natural catalytic autoantibodies (abzymes) posessing proteolytic (protoabzymes) and DNA-hydrolysing (DNA-abzymes) activities are detectable in clinical and experimental autoimmune disorders. Catalytic activity of abzymes is revealed in the patients with different forms of systemic (nonspecific) and organ-specific autoimmune pathology. The opportunity of assaying the abzymes for intensity monitoring of autoimmune disease is also described.

Abstract. The goal of our research was comparative study of the most important parameters of subset cytoarchitectonics in the patients with the different courses of myocarditis and evaluation of their pathogenetic and clinical value in the practice of the physician. We have investigated 99 patients with myocarditis and 40 healthy donors. In patients with malignant course of disease we revealed increased activation index of T/B-cells; increased expression of the activation markers on the both lines of differentiation; disproportion in the immunoregulatory subsets with increased role of dendric cells; decreased intensity of the autoreactive T-cells apoptosis. in the patient with the In patients with nonmalignant course of disease expressed signs of immunopathology were not found. Thus, study of activation markers on the cells of the peripheral blood is more informative and noninvasive method of diagnostics of myocarditis.