Purpose of the study. To evaluate the cellular, genomic (gene copy number) and transcriptomic (gene expression) effects of P.hybridus (L.) secondary metabolites when they affect the HeLa cell line.Materials and methods. The isolation of secondary metabolites from plant material and its identification were carried out by preparative chromatography. The composition was determined using mass spectrometric analysis, and the final verification of structural formulas was carried out by nuclear magnetic resonance at the Department of Natural Compounds, the Faculty of Chemistry of the Southern Federal University. The subsequent phase of the study was conducted using both cultural and molecular methods. HeLa cells were cultivated under standard conditions in a MEM medium. Once the confluence level was reached 75–80 %, the nutrient medium was replaced with the introduction of the studied compounds (at a concentration of 4 micrograms/ml) and cultivated for 72 hours. Cell mortality was determined using a NanoEnTek JuliFl counter (Korea) in the presence of 0.4 % trypan blue. The assessment of apoptosis following secondary metabolite exposure was conducted on a BD FACSCanto II flow cytometer using the FITC Annexin V Apoptosis Detection Kit I. The level of replication and expression of the genes responsible for apoptosis was assessed by digital droplet PCR (ddPCR).Results. The following compounds were isolated and verified, and were assigned the following sequence numbers to facilitate their use in the experiment: No. 2 – 2,4-dihydroxy-2,5-dimethylfuran-3(2H)-one, No. 3 – 5-(hydroxymethyl) furan-2-carbaldehyde, No. 5.3 – 2,2,8-trimethyldecahydroazulene-5,6-dicarbaldehyde, P. hybridus (L.) At the stage of cell death assessment, it was found that the greatest effect was achieved in the compound under ordinal No. 2. However, the evaluation of the copy number and expression of the CASP8, CASP9, CASP3, BAX, BCL2, TP53, MDM2, CDKN1B, CDK1, CCND1, CCND3, and RB1 genes by DD-PCR revealed the presence of apoptosis initiation in tumor cells at the molecular level under the action of compounds No. 2 and No. 5.3 obtained from P. hybridus (L.).Conclusion. The outcomes were multifeatured. Only compound 2,4-dihydroxy-2,5-dimethylfuran-3(2H)-one exhibited a pronounced cytostatic effect out of all compounds utilized in the experiment. Concurrently, the compound 2,2,8-trimethyldecahydroazulene-5,6-dicarbaldehyde was found to induce an increase in the expression of the CASP3, CASP8, TP53, and BAX genes.

УДК 616.24-006.66 ОСОБЕННОСТИ ПОПУЛЯЦИОННОГО СОСТАВА ИММУНОКОМПЕТЕНТНЫХ КЛЕТОК ПЕРИФЕРИЧЕСКОЙ КРОВИ У ПАЦИЕНТОВ С НЕМЕЛКОКЛЕТОЧНЫМ РАКОМ ЛЕГКОГО НА ФОНЕ ПРОВОДИМОЙ ИММУНОТЕРАПИИ С РАЗЛИЧНОЙ ЕЕ ЭФФЕКТИВНОСТЬЮСагакянц А

Purpose of the study. Was to reveal characteristics of the immunocompetent cells in colon cancer (CC) according to the disease stage, and to identify prognostic factors of cancer development.Materials and methods. The study included 50 patients with CC: stage I – 4 patients (8 %), II – 25 (50 %), III – 21 (42 %). All patients underwent standard surgical intervention at the initial stage, the obtained material was used for subsequent studies: a cell suspension was obtained from the tumor tissue, peritumoral zone (1–3 cm from the tumor), which was treated with an antibody panel (Becton Dickinson, USA) to identify the main subpopulations of leukocytes and lymphocytes.Results. The tumor tissues of patients with stages I + II showed a decrease in the relative number of DP, DN, NKT and CD19+, compared to peritumoral tissues, by 33 %, 42 %, 31 % and 82 % respectively. Tumor tissues of stage III patients demonstrated elevated relative numbers of CD3+, CD4+, NK by 57 %, 34 %, 48 %, and decreased DP, DN, NKT, CD19+ by 33 %, 74 %, 31 %, 59 %, compared to peritumoral tissues. DP, DN, NKT and CD19+ in the tumor decreased by 78 %, 74 %, 39 %, 60 %, respectively, and the relative number of lymphocytes increased by 138 %, compared to the tissues of the resection line. A comparative analysis of local immunological parameters in the tumor tissues of patients with СС revealed that the relative numbers of lymphocytes and CD19+ were 58 % and 87 % higher, and DP and DN were 33 % and 27 % lower in tumor tissues of stage III patients, compared to tumor tissues of stage I + II patients.Conclusion. Thus, the obtained features of the local population and subpopulation composition of immunocompetent cells in CC, depending on the stage of the tumor process, can be used to predict the clinical course of the disease.

Purpose of the study. To isolate and verify pure fractions of secondary plant metabolites contained in B. vulgaris (L.) and P. hibridus (L.), as well as to conduct a model experiment and molecular genetic study to evaluate their cytotoxic effect in vitro on the HeLa cell line.Materials and methods. The isolation and verification of all compounds used in the experiment were carried out using column chromatography and nuclear magnetic resonance methods at the Department of Natural Compounds, Faculty of Chemistry, Southern Federal University. Subsequently, an experiment was conducted using cultural and molecular methods on the HeLa cell line in three repetitions for each test compound; after incubation with them, the numbers of dead cells were counted on the automated NanoEnTek JuliFl counter, and the numbers of cells in apoptosis were measured by flow cytometry on the BD analyzer FACS Canto II. The level of copy number variation and expression of genes responsible for apoptosis were assessed by real-time PCR (RT-PCR). In total, three substances were studied, with two concentrations (4 and 12 μg/ml) and two exposures (24 and 72 hours) for each of them.Results. In the first stage of the study, we isolated and verified the berberine alkaloid extracted from the roots of B. vulgaris (L.), as well as 2,4-dihydroxy-2,5-dimethylfuran-3(2H)-one and 2,2,8-trimethyldecahydroazulene-5,6-dicarbaldehyde from P. hibridus (L.). The subsequent stage of the study demonstrated the maximal cellular death under the action of berberine at a 72-hour exposure. However, the RT-PCR assessment of the copy number variation and expression of the CASP8, CASP9, CASP3, BAX, BCL2, TP53 and MDM2 genes revealed the presence of apoptosis initiation in tumor cells at the molecular level under the action of all the studied compounds: both berberine and furan and azulene derivatives derived from P. hibridus (L.).Conclusion. All compounds used in the experiment exhibited a cytotoxic effect on the HeLa cell line. Berberine alkaloid showed the most pronounced cytotoxic effect on the HeLa line as recorded by all methods used in the study. Terpenoids 4-dihydroxy-2,5-dimethylfuran-3(2H)-one and 2,2,8-trimethyldecahydroazulene-5,6-dicarbaldehyde, when exposed to the HeLa line, caused an increase in the copy number variation and expression of the CASP9, CASP3 loci, which are among the main activators of apoptosis. They also influenced the expression of TP53 and MDM2 loci.

ПАРАМЕТРЫ СИСТЕМНОГО КЛЕТОЧНОГО ИММУНИТЕТА У ПАЦИЕНТОВ С МЕЛАНОМОЙ КОЖИ НА ФОНЕ ПРОВОДИМОЙ ИММУНОТЕРАПИИ С РАЗЛИЧНОЙ ЕЕ ЭФФЕКТИВНОСТЬЮСагакянц А.Б. 1 , Теплякова

The purpose of this study was to investigate the numbers of cells with CSC markers in samples of human colorectal cancer (CRC) xenografts isolated in animals with tumors after exposure to the small molecule XAV-939, 5-fluorouracil, and to the combination of XAV-939 with 5-fluorouracil. The model was created in Balb/c Nude mice. Tumor fragments of human CRC were transplanted into adipose tissue of animals under the skin of the right thigh. The fourth xenograft generation was used for the study. Animals were divided into groups receiving XAV-939, 5-fluorouricil, their combination, and the control group. Tumor nodes were measured twice a week; at the end of the experiment, the number of cells with the CSC phenotype was determined in the tumor material of the animals. By the end of the experiment, tumor nodules in the group of animals receiving the combination of drugs were significantly smaller than in the control group. The highest levels of cells with phenotypic signs of CSCs were characteristic of samples from the control group, and for samples obtained from the group receiving 5-fluorouracil as monotherapy. Decreased numbers of these cells were observed in groups receiving XAV-939 and the combination of XAV-939 and 5-fluorouracil.

Purpose of the study. To study the features of the local distribution of populations of immune system cells in patients with non-invasive muscular bladder cancer of various degrees of malignancy.Materials and methods. The study included 51 patients with newly diagnosed non-muscle-invasive bladder cancer (papillary urothelial carcinoma) who received complex treatment and follow-up after 9 months at the oncourological department of the National Medical Research Center of Oncology. Patients were divided into two groups: group 1 – with a tumor of low malignant potential (Low grade – LG), n = 31; group 2 – with a tumor of high malignant potential (High grade – HG), n = 20. After 6–9 months, 24 patients were diagnosed with a relapse of the disease – in 48,4 % in patients of group 1 (n = 15) and in 45 % – in group 2 (n = 9). In cell suspensions obtained from the primary and recurrent tumors, as well as the perifocal zone, the relative number of populations of immunocompetent cells was estimated using flow cytometry. A comparison was made of the content of individual populations of lymphocytes in the tumor tissue, the perifocal zone of primary and recurrent lesions of various degrees of malignancy. Statistical processing was performed using Statistica 13.0.Results. The development of a recurrent tumor of low malignant potential is accompanied by the involvement of cells of innate immunity (NK- and NKT-lymphocytes) into its microenvironment, which is associated with an imbalance in the number of main cells of adaptive immunity – a fairly pronounced decrease in the tumor of T-lymphocytes of the helper-inductor type was noted with a constant content cytotoxic T-lymphocytes, as well as the multidirectional nature of changes in DP- (decrease) and DN-lymphocytes (increase). A feature of the development of a recurrent tumor of high malignant potential is that it is accompanied by the involvement of innate immunity cells (NK- and NKT-lymphocytes) into its microenvironment, as well as multidirectional changes in DP- (decrease) and DN-lymphocytes (increase).Conclusion. Studies of the population composition of tumors and their perifocal tissues of NMIBC revealed a number of features that are reflected in the redistribution of cytolytic cells, the formation of immunosuppressive conditions, which are reflected both in the manifestation of the biological properties of tumor cells and in changes in the cellular composition of bladder tissues involved in the process. development and progression of cancer.

УДК 616-006.66ОСОБЕННОСТИ ПОПУЛЯЦИОННОГО СОСТАВА ИММУНОКОМПЕТЕНТНЫХ КЛЕТОК В ОПУХОЛИ ПРИ МЫШЕЧНО-НЕИНВАЗИВНОМ РАКЕ МОЧЕВОГО ПУЗЫРЯ РАЗЛИЧНОЙ СТЕПЕНИ ЗЛОКАЧЕСТВЕННОСТИ И РЕЦИДИВИРОВАНИИ ЗАБОЛЕВАНИЯ Сагакянц А.Б. 1 , Белякова Л.И. 1 , Шевченко А.Н. 1 , Бондаренко Е.С. 1 , Златник Е.Ю. 1 , Новикова И.А. 1 , Филатова Е

Purpose of the study. To evaluate the number of cells with the CD45+/- phenotype expressing Toll-like receptors (TLRs) in tissues of the tumor, peritumoral area and resection line tissues in colon cancer (CC) with various tumor locations.Materials and methods. The study included 50 patients with CC. All patients underwent surgery as the primary treatment, and tissue material was collected from the patients. Expression of TLRs (2, 3, 4, 8, 9) on CD45+, CD45- cell populations was determined by flow cytometry in cell suspensions obtained from tissues of the tumor, peritumoral area (1–3 cm from the tumor) and resection line tissues (~10 cm from the tumor) with further calculation of the percentage of cells with the corresponding phenotype from the total number of cells.Results. An analysis of left-sided colon tumors showed lower percentage of CD45- cells expressing TLR4, 8, compared to rightsided tumors, by 38 % and 25 %. A comparative analysis of the number of CD45+ cells expressing TLR 2, 4 showed their decrease by 81 % and 87 %, respectively, compared with right-sided tumors. An assessment of the data in the perifocal zone of left-sided colon tumors, compared with right-sided ones, demonstrated a decrease in the percentage of cells with the CD45- phenotype that express TLR4, by 61 %. Resection line tissues in left-sided tumors, compared with right-sided ones, showed a statistically significant increase in the percentage of CD45- cells that express TLR 2, 4 by 205 % and 55 %, respectively. The number of CD45+ cells expressing TLR 4 decreased by 87 %. An assessment of the number of cells expressing TLRs 3 and 9 in the tumor, peritumoral area and resection line tissues did not reveal significant differences.Conclusions. Lower number of cells with CD45+ and CD45- phenotypes express TLRs 2, 4, 8 in left-sided colon tumors and their peritumoral tissues, compared to right-sided cancer.

Background: Colorectal cancer is now an urgent problem in oncology. Recently, specialists have been interested in a comparative analysis of differences in the clinical course of malignant tumors in the proximal and distal colon. The sections differ not only in their embryogenesis and sources of blood supply, but also in the clinical course and population and epidemiological characteristics. The issue of distinctive immunological characteristics of tumors of the colon depending on the location remains open.Objective: A comparative analysis of local subpopulations of immunocompetent cells and an assessment of number of cells with the CD45+/- phenotype expressing toll-like receptors (TLRs) depending on the tumor location on the right or left sides of the colon.Material and methods: The study included 50 patients with verified colon cancer. The majority of patients were females – 26 (52%), aged 67 ± 0.4 years, and 50% of patients with stage II disease. Depending on the tumor location (the right or left sides of the colon), the patients were divided into 2 groups of 25 people each. All patients underwent standard surgery at the initial stage. The obtained material was used for subsequent studies: a cell suspension was obtained from a tumor tissue fragment, the perifocal zone (1–3 cm from the tumor) which was processed using an antibody panel (Becton Dickinson, USA) to identify the main subpopulations of leukocytes and lymphocytes. Expression of TLRs (2, 3, 4, 8, 9) on CD45+, CD45- cell populations was also determined using the BD FACSCanto flow cytometer (Becton Dickinson, USA). Statistical processing of the results was performed using the STATISTICA 13.3 package (StatSoft Inc., USA).Results: A comparative analysis of immunological parameters, depending on the tumor location on the right or left sides of the colon, showed:Tissues of the right-sided tumors had a higher T-lymphocytic infiltration, compared to the left-sided tumors, while the latter showed a higher B-lymphocytic infiltration (p = 0.025).Peritumoral zone tissues of left-sided tumors demonstrated a decrease of lymphocytes levels (p = 0.027), NKT – (p = 0.035), NK – (p = 0.041) and В lymphocytes (p = 0.038), and a significant increase in CD8+- (p = 0.02) and DP cells (p = 0.0018).Left-sided tumors showed a percentage decrease of CD45- cells expressing TLR4 and TLR8, compared to right-sided tumors, by 38% (p = 0.038) and 25% (p = 0.043).There was a decrease in the number of CD45+ cells expressing TLR2 and TLR4 in left-sided tumors by 54% (p = 0.035) and 33% (p = 0.04) respectively, than in right-sided tumors.The percent of CD45- cells expressing TLR4 in the perifocal tissues of left-sided tumors decreased by 61% (p = 0.031) in comparison to the corresponding tissues in right-sided tumors.The numbers of CD45+ cells expressing TLR2 and TLR4 were 81% (p = 0.02) and 87% (p = 0.018) lower respectively in the peritumoral tissues of left-sided tumors, compared to the corresponding tissues in right-sided tumors.Conclusion: The revealed characteristics of the local subpopulations of immunocompetent cells and the numbers of CD45+/- cells expressing TLRs depending on the tumor location on the right or left sides of the colon can serve as a prognosis of the disease clinical course and the choice of further treatment tactics.

Clinical prognosis in malignant tumors` is among the most challenging problems of contemporary medicine. It is thought to depend on both biologic properties of tumor cells and patients` immune status. The features of tumor cells and immune reactions are closely interrelated and mutually conditioned. Therefore, possible application of their characteristics as prognostic markers is of great fundamental and clinical importance. The aim of our study is to find out the most significant immune factors for prognosis of colorectal cancer (CRC) based on estimation of local and system immune factors, and some characteristics of tumor cells in the patients at various stages of the disease and different clinical course. Cellular factors of immunity and cytokines were studied in blood and tumor tissue of 299 patients with colorectal cancer (stages I-IV). Malignant cells expressing stem cell markers CSC), MHC and PDL-1 molecules were also counted in the tumor tissue. Blood samples were drawn prior to operation, and tissue samples were taken during surgery being the 1st line of treatment. Flow cytometry techniques (FCM), immunohistochemistry (IHC), and ELISA approach were employed. We have compared data on the patients at different CRC stages (with or without local and distant metastases), as well as cases with different course of the disease (evolving distant metastases and fatal outcome during period of observation). Our results demonstrated increased amounts of NK-cells and IL-6 concentration, along with decreased percentage of blood CD4+ cells in the patients with local metastases, as well as higher CSC numbers in malignant tissue. The initially generalized CRC cases with distant metastases were characterized by high levels of blood IL-6, monocytes and granulocytes responding to fMLF, while in tumor tissue elevated amounts of NKT, CSC and decreased expression of MHC and PDL-1 were observed on tumor cells, like as lower PD-1/ PDL-1 expression on tumor-infiltrating lymhocytes. Unfavorable CRC dynamics, i.e., metastasizing during the observation period was preceded by increased levels of IL-10 in blood, NK cells with poor cytotoxicity, monocytes and granulocytes responding to fMLF. In tumor tissue, overexpression of CSC markers and hypo-expression of MHC on tumor cells were noted. Fatal outcome was preceded by elevation of blood IL-6levels, tissue levels of NKТ and CSC percentages, along with decreased NK cells subset (CD16dimCD56bright) in blood, and decline of MHC-expressing cells in the tumor. Thus, high blood levels of IL-6 and IL-10, fMLF-responding monocytes and granulocytes, as well as elevated amounts of NKT and CSC, hypo-expression of MHC in tumor tissue could be considered prognostic markers of unfavorable course in CRC patients. Decrease of PD-1/PDL-1 expression on tumor cells and lymphocytes from its microenvironment in advanced CRC is of special attention, because checkpoint inhibitors are prescribed in such cases.

Purpose of the study. Determine the content of cancer stem cells (CSCs) in the tumor tissue (TT) and perifocal tissues (PT) in muscle-non-invasive bladder cancer.Materials and methods. We’ve examined fragments of TT and PT of 7 muscle-non-invasive bladder cancer (NMIBC) after surgical intervention – transurethral resection of the urinary bladder (TUR). In tissue samples that were used to obtain cell suspension of TT and PT using the BD Medimachine apparatus (BD, USA) was treated with monoclonal antibodies CD45-APCCy7, CD44-FITC, CD133-РЕ, CD24-PE (BD, USA) and were assessed on flow cytometer FacsCantoII (BD, USA). The percentage of cells with CSC phenotypic markers was determined in the analysis sample: CD45-CD44+CD24+, CD45-CD44+, CD45-CD24+, CD45-CD133+, CD45-CD44+CD133+. The presence of significant differences in the groups was evaluated using the STATISTICA 13 software package and the differences between the samples were considered significant at p < 0.05. The percentage of cells of the corresponding phenotype was calculated relative to the total number of cells. The percentage of cells with the corresponding phenotype was calculated relative to the total number of cells.Results. The relative numbers of cells with CSC phenotypic markers, such as CD24, CD44, were 77 % and 58 % higher in TT than in PT: 18.3 ± 3.5 vs. 4.3 ± 2.1, p ≤ 0.044 and 15.5 ± 5.3 vs. 6.5 ± 0.8, p ≤ 0.043, respectively. The number of CD133+ cells was 83 % higher in PT compared to TT – 41.6 ± 12.1 vs. 22.7 ± 7.6, p ≤ 0.047.Conclusion. The study of CSCs is a promising direction for the study of oncogenesis and can be used to assess the nature of the further development of relapse and / or progression of the disease, as well as various therapeutic approaches that are aimed at eliminating with CSC phenotypic markers and blocking the pathways leading to the emergence and maintenance of this cell population in patients with NMIBC.

УДК 616-006.66ОСОБЕННОСТИ ЛОКАЛЬНОГО КЛЕТОЧНОГО ИММУНИТЕТА ПРИ РАКЕ ОБОДОЧНОЙ КИШКИ В ЗАВИСИМОСТИ ОТ ЛОКАЛИЗАЦИИ ОПУХОЛЕВОГО ПРОЦЕССА Кит О.И. 1 , Дженкова Е.А. 1 , Мирзоян Э.А. 1 , Сагакянц А

In this work we studied the effect of different concentrations of interleukin-2 (IL-2) and interferon-γ (IFN-γ) on the proliferation and immunophenotype of lymphocytes obtained from patients with locally advanced breast cancer (stage II-III) after immunomagnetic depletion of regulatory T cells (Tregs) from the general pool of lymphocytes in vitro. The peripheral blood of 11 patients was used as a material. Peripheral blood mononuclear cells (MNCs) were isolated and regulatory T lymphocytes were removed by immunomagnetic separation. After separation, the cells were cultured in a nutrient medium RPMI-1640 with 10% fetal bovine serum (FBS) for 7 days. Lymphocytes were activated on the first day of cultivation with one of the following cytokines: IFN-γ (10 IU/mL), IL-2 (0.1 or 1 μg/mL); IL-2 (0.1 or 1 μg/mL) and IFN-γ together. Lymphocytes without cytokine addition was used as a control. Cells were counted using an automatic counter before addition cytokines and after 2, 4, and 7 days of cultivation with cytokines, and their phenotype was examined. The results showed some phenotypic differences in some elements of cellular immunity between control and experimental samples. Particular attention is paid to the description of changes in the expression of surface markers in the natural killer (NK) subpopulation. It was noted that the proportion of Tregs, despite their preliminary depletion, increased after exposure to cytokines. As a result, the preliminary decrease in the proportion of Treg cells before the stimulation of lymphocytes did not produce the expected effect, therefore the use of depletion in such a methodological mode did not lead to significant results. However, one should not exclude the possibility of inhibiting Tregs by other methods.

Purpose of the study. Evaluation of the cytotoxic effect of strains RVK100 and RVK228 of a new unclassified group of human rotaviruses on human peripheral blood mononuclear cells in vitro. Materials and methods. As a material for the study, we used peripheral blood mononuclear cells of a healthy donor. The cells were exposed to two strains of rotaviruses RVK100 and RVK228 for 24 and 48 hours. The cytotoxicity of the tested viruses was assessed using the Colorimetric Cell Viability Kit I (WST-8) (PromoCell, Germany). Analysis of lymphocytes subpopulation composition was assessed on a FACSCantoII flow cytometer (BD, USA) using monoclonal antibodies to human antigens: CD3, CD4, CD8, CD16/56, CD19, CD45, CD38, HLA-DR. Results. According to the cell viability test, there was no significant decrease in the number of living cells in the samples with the addition of viruses in comparison with the control. On the contrary, after 48 hours of cultivation in the samples with the addition of RVK228, the number of living cells was significantly higher than in the control. The study of lymphocytes subpopulation composition showed a relative increase in the number of early activation markers on T cells in samples with viruses, which was also more pronounced in samples with the addition of RVK228. Conclusion. The investigated strains of rotaviruses have no cytotoxic effect on human peripheral blood mononuclear cells. Moreover, the RVK228 strain is likely to have the ability to activate lymphocytes.

Introduction.The actual task of modern adoptive cancer immunotherapy is the selection of the optimal composition of cytokines for ex vivo stimulation of immunocompetent cells for subsequent administration to oncological patients.The purpose of the studywas to compare the effect of interleukin (IL) 2, 7, 15 and their combinations on the proliferation of natural killer cells in breast cancer (BC) patients in vitro.Materials and methods.The research was conducted on natural killer cells isolated by magnetic separation from mononuclear cells of peripheral blood of ten patients with locally advanced BC (stage II). After separation, the cells were cultured at a concentration of 2.5 × 105 cells / ml for 10 days in RPMI 1640 medium supplemented with cytokines at a concentration of 40 ng / ml each in five experimental variants: IL‑2; IL‑7; IL‑15; IL‑7 / IL‑15; IL‑2 / IL‑7 / IL‑15. On the 10th day of cultivation, the phenotype of cells and the cell cycle were studied by flow cytometry. For immunophenotyping of cells, we used monoclonal antibodies to antigens: CD3, CD16 / 56, CD45, CD4, CD19, and CD8. For cell cycle study cells were stained with propidium iodide.Results. On the final 10th day of cultivation the number of living cells expressed as percentage of the seeding numbers were significantly different from control (45.9 %) in samples IL‑2 (86.8 %) and IL‑7 / IL‑15 (85.6 %), IL‑15 (76.4 %), IL‑2 / IL‑7 / IL‑15 (75.8 %). The proportion of natural killers (CD16+CD56+) significantly differed from the control (18.2 %) in samples IL‑2 (45.6 %), IL‑15 (39.9 %), IL‑7 / IL‑15 (36.2 %), IL‑2 / IL‑7 / IL‑15 (35.9 %). The propor‑ tion of natural killer T cells (CD3+ / CD16+CD56+) significantly differed from the control (0.4 %) in samples incu‑ bated with IL‑2 (2.06 %), IL‑15 (2.2 %), IL‑7 (0.9 %), IL‑7 / IL‑15 (1.26 %), IL‑2 / IL‑7 / IL‑15 (2.46 %). All experimental tests also showed a significant increase in the proportion of cells in the S‑phase and increase in the proliferation index (G2 / M + S).Conclusion.The maximum stimulation of the proliferation of natural killer cells isolated from the blood of patients with BC in vitro was obtained by stimulation with IL‑15 alone and in combinations with γc‑cytokines.

Glial tumors comprise about 60 % of primary malignant brain tumors, and 70 % of them show morphological signs of high-grade cancer (High Grade Gliomas III , IV WHO 2016) [1, 2]. Despite a significant technical pre- and intraoperative progress as well as advances in radiotherapy and chemotherapy, the overall median survival is very low, being less than 20 months [3] and less than 12 months in patients with relapse [4]. Recent studies have shown that chemo- and radioresistance is due to the existence of cancer stem cells [5, 6]. Poor treatment outcomes require the development and implementation of new approaches to the treatment of highgrade gliomas. In recent years, increasing attention has been paid to the development of immunotherapeutic treatment approaches, including the development of oncolytic virotherapy. Tropism to target cancer cells, as well as various viral vectors, has been developed using methods of genetic engineering; synergism of viruses and adjuvant therapy has been studied. Despite extensive experimental studies of the mechanism of oncolysis [1], there are only a few reports on Phase I–II clinical trials. This review considers the most successful applications of oncolytic viruses in relation to glioblastoma in animal models and their translation into clinical practice in patients.

ОПУХОЛЕВЫЕ СТВОЛОВЫЕ КЛЕТКИ РАКА ЯИЧНИКОВ В МЕХАНИЗМАХ УСТОЙЧИВОСТИ К ХИМИОТЕРАПИИ Сагакянц А.Б. 1 , Якубова Д.Ю. 1 , Моисеенко Т.И. 1 , Златник Е.Ю. 1 , Меньшенина А.П. 1 , Жукова Г.В. 1 , Вереникина Е.В. 1 , Ульянова Е.П. 1 , Бондаренко Е