Novel coronavirus disease 2019, caused by the SARS-CoV-2, initiate humoral and cellular immune responses against diverse virus antigens. The assessment of SARS-CoV-2-specific is mainly carried out in routine practice by determining specific immunoglobulins. However, high variability in S-protein structure in new genovariants of SARS-CoV-2 virus and the lack of correlation between specific antibodies and CD8+ T-lymphocytes underlie false negative responses, and mass assessment of cellular immunity is complicated due to the complexity of applying ELISPOT and cytofluorometry techniques. To solve this issue, a diagnostic method was developed for assessing SARS-CoV-2-specific cellular immune response, which is based on a skin test followed by evaluating a delayed-type hypersensitivity reaction involving antigen-specific memory T-lymphocytes. A diagnostic preparation CoronaDerm-PS is based on Cord_PS, which is a hybrid recombinant protein consisting of parts of the SARS-CoV-2 structural proteins S, M, N, E. The specific activity of this chimeric antigen was analyzed in cultured T-lymphocyte activation test by assessing interferon-γ production using cytofluorometry. To investigate the chimeric antigen specific activity, a preclinical safety study with CoronaDerm-PS preparation in experimental animals was conducted. A dose-dependent developing skin reaction was observed in 90–100% of guinea pigs vaccinated by EpiVacCorona, CoviVac, Gam-COVID-Vac, which confirms a potential for assessing post-vaccination cellular immunity using CoronaDerm-PS preparation. Upon this, the presence of functionally active T-cell-antigenic epitopes in the recombinant polypeptide allows to evaluate SARS-CoV-2-specific response illustrated by detected response after Gam-COVID-Vac (S-protein) and EpiVacCorona (N-protein) vaccination. Thus, a skin test based on CoronaDerm-PS preparation may be a promising diagnostic tool for rapid mass screening requiring no specialized laboratory equipment for assessing populational SARS-CoV-2-specific immunity. Such a test is distinguished by advantages such as ease of analysis, high specificity and sensitivity. The final decision-making on using this test in a real-world practice may achieved after conducting further clinical safety and effectiveness trials.

Granulocyte-macrophage colony stimulating factor (GM-CSF) is a myelopoietic growth factor that exerts pleiotropic effect not only on the differentiation of immature progenitor cells into polymorphonuclear neutrophils, monocytes/macrophages and dendritic cells, but also controls the functioning of differentiated cells. GM-CSF is currently being investigated in clinical trials as an immunomodulator and adjuvant. However, a wide range of biological activities and, sometimes, paradoxical effects of this cytokine require more thorough studies of its action, in order to predict its efficacy under different conditions of immunotherapy. In this work, we have studied the effect of recombinant human GM-CSF on metabolic activity of mouse peritoneal exudate cells in primary cell cultures. Metabolic (redox) activity of the cells was assessed by their ability to reduce nitroblue tetrazolium (NBT) in the course of MF- and Fc-dependent phagocytosis triggered by addition of opsonized zymosan, or sheep erythrocytes to the culture medium. We have shown the dose-dependent stimulatory effect of GM-CSF on the oxidative metabolism of phagocytic peritoneal macrophages and neutrophils. Upon culturing the pepton-elicited cells at wide range of GM-CSF concentrations (5 to 40,000 ng/mL) for 2 and 24 hours, a more pronounced effect of the substance was observed for neutrophils. The GM-CSF preparation caused a significant increase (by 13-17%) in the redox activity of neutrophils induced by opsonized zymosan that persisted at a low dose range, and was retained after 24 hours. The stimulatory effect of GM-CSF on macrophages with NBT index increase by 16% was observed in the short-term cultures. In general, the elicited cells of both types showed a more pronounced response to lower concentrations of GM-CSF (5-125 ng/mL), and weaker effect at higher doses of the preparation. A similar dependence was found when studying the resident macrophages. Culturing of resident cells with GM-CSF at the doses of 5,000 to 40,000 ng/mL for 24 hours caused a significantly increased redox activity of the cells induced by zymosan, or sheep erythrocytes (by 33-52%). In both cases, the maximal response was detected at a dose of 5,000 ng/mL and decreased with increasing dose. The stimulatory effect of GM-CSF upon resident macrophages was more pronounced as compared to elicited cells, which was characterized by the prolonged period of cell activation (up to 24 hours of culture). The data obtained are of interest, in view of prospective usage of GM-CSF as a component of immunomodulatory and adjuvant therapy for various infectious diseases.

It was established that recombinant human tumor necrosis factor alpha in a delivery system has pronounced antitumor and immunomodulating effects in murine B16-F10 melanoma model. The preparation significantly inhibited the growth of experimental tumor (TGI 60%), increased tumor infiltration by immune system cells (CD3+, CD11b+) and increased the activity of immunocompetent cells mediating antitumor response.

The evaluation of adaptogenic properties of lyophilized immunoglobulins Y isolated from chicken yolks was performed with biological testing methods (IgY). In animal experiments, in the forced swimming test (FST), a prior five- and ten-fold oral administration of yolk chicken IgY in a dose of 2 mg per mouse led to a relevant increase in animal endurance.

Introduction. Tumor necrosis factor α (TNF-α) is a natural cytokine, characterized by pronounced antitumor properties. A wide range of side effects serves as an obstacle for the use of TNF-α in clinical practice. One of the ways to improve its therapeutic properties is to increase the tropism of the cytokine to the tumor tissue by incorporating it into the targeted delivery system.The aim of the study was to evaluate the antitumor activity of the preparation containing TNF-α as part of the artificial “virus-like particle” (VLP-TNF-α), developed in SRC VB “Vector” as a transport system for delivering proteins to target cells.Materials and methods. The antitumor effect of VLP-TNF-α preparation was evaluated in experimental B16F10 melanoma model by the change of dynamics of tumor growth (volume, mass) and its morphological structure (presence of necrotic processes, blood vessel destruction). The number of the effector immune cells (CD3+, CD11b+) in the tumor tissue was determined by immunohistochemical method.Results. It has been shown that VLP-TNF-α administered intravenously at the doses of 5 × 104 and 1 × 105 IU/mouse inhibits the growth of the primary tumor. The most pronounced and stable effect was observed with a five-fold administration at the dose of 1 × 105 IU/mouse every other day: tumor growth inhibition was 40 % on the 1st day, and 47 % on the 7 th day upon the treatment. Injections of the preparation resulted in the increase of necrosis number, destruction level of the tumor tissue, development of damage and destruction of the tumor blood vessels and its infiltration with immunocompetent cells.Conclusion. The obtained data indicates that TNF-α within the delivery system exerts antitumor activity, which suggests the possibility of its further use for the treatment of malignant neoplasms, in particular, melanoma.The study was performed in accordance with ethical principles adopted by the European Convention for the protection of vertebrate animals used for experimental and other scientific purposes.

Granulocyte-macrophage colony-stimulating factor (GM-CSF) occupies an important place in the processes of hematopoiesis and immunogenesis, and its biological effects are extremely diverse. GM-CSF is a natural cytokine capable of inducing the differentiation, proliferation and activation of a significant number of populations of immunologically active cells. Due to its immunotropic effects and the ability to activate processes of blood cell forming, GM-CSF, like other colony-stimulating factors, has found a fairly wide medical application as hemostimulant and immunocorrector for various pathological conditions. There is reason to suppose that the immunomodulating properties of GM-CSF preparations can also be used to increase the efficiency of low immunogenicity vaccines, when vaccinating patients suffering from immunological deficiency. This review covers the results of recent studies on the immunoadjuvant effects of GM-CSF in vaccination against infectious diseases, for enhancing the effeciency of engineered vaccines based on protein antigens, as well as a genetic adjuvant for DNA vaccination.

—The review summarizes literature data on the development of drugs based on natural and synthetic high-polymeric double-stranded RNA (dsRNA), their antiviral, immunoadjuvant, and antitumor properties. Special attention is paid to cell receptors responding to exogenous dsRNA, pathways of dsRNA-dependent antiviral reaction, ability of dsRNA to inhibit growth and induce apoptosis of malignant cells. It has been shown that enhancing the innate immune response with dsRNA can be an effective component in improving methods for treating and preventing infectious and cancer diseases. The further use of dsRNA for the correction of pathological processes of different origin is discussed.

Effect of alkoxyamines on normal and tumor cells was studied in vitro and in vivo. In vitro experiments showed that alkoxyamines produce a dose-dependent toxic effect on cells of human breast tumor MCF7 line. Transplantation of Krebs-2 ascites carcinoma cells preincubated with alkoxyamines to mice did not induce tumor growth. An opposite effect was observed in normal mouse cells: functional activity of peritoneal macrophages increased. The possibility of using alkoxyamines as theranostic agents is discussed.

It is shown in experiments in mice that the double-stranded RNAs (dsRNA) of bacteriophage ф6 has pronounced interferon-inducing properties and the ability to enhance the functional activity of phagocytes. The maximum level of IFN-а in serum of outbred ICR mice was observed 5 h after the i.p. administration of the phage dsRNA at doses of 0,8 and 8 mg/kg, and the number of NBT-positive peritoneal macrophages increased 5 – 24 h after the administration. DsRNA of bacteriophage ф6 under conditions of three-fold administration (3 h before, 1 and 3 days after infection) increased the average life span and the number of surviving mice infected with influenza virus (IV) A/Chicken/Kurgan/05/2005 (H5N1). The IV titers in lung homogenates of dsRNA-administered mice were significantly lower on 4th and 6th days after the infection with IV A/Bishkek/03/2009 (H1N1)pdm09 than in untreated animals. The antiviral effects of the phage dsRNA were comparable to those of Tamiflu. These results give the future outlook for the development a new drug based on the dsRNA of bacteriophage ф6 as a preventive and therapeutic medicine for viral infections.

The method of obtaining and purification of the double-stranded RNA of bacteriophage p6 was developed. It was shown that the created method makes it possible to obtain high purity dsRNA preparations possessing interferon-inducing and phagocyte-stimulating activity.

В культуре клеток меланомы B16-F10 проведено сравнительное изучениецитотоксической активности рекомбинантного человеческого фактора некрозаопухолей (рчФНО-альфа) и его новых форм в виде конъюгатаи молекулярной конструкции, содержащих алендроновую кислоту, а такжевозможности модуляции противоопухолевых эффектов ФНО-альфа рекомбинантныманалогом интерферона гамма человека. Продемонстрирована возможность усиленияцитотоксического действия рчФНО-альфа внизких дозах при совместном использовании с интерфероном гамма. Установлено, что рчФНО-альфа в составеконъюгата и молекулярной конструкции сохраняет свои антипролиферативныесвойства. Молекулярная конструкция,содержащая ФНО-альфа, дсРНК и алендроновую кислоту, проявляет более выраженноецитотоксическое действие в отношении клеток меланомы по сравнению с ФНО-альфа.

Проведено исследование иммуномодулирующей активности аналога интерферона гамма дельтаферона на мышах ICR с адъювантным артритом. Показано, что трехкратное введение дельтаферона ингибирует развитие локальной воспалительной реакции, вызванной введением полного адьюванта Фрейнда. Дельтаферон снижал метаболическую активность перитонеальных макрофагов, пролиферативную активность лимфоцитов селезенки в острой фазе воспалительного процесса.

AIM:Evaluation of composite formulation of yeast double stranded RNA with polyglucinum (dsRNA-PG) effect on non-specific antiviral resistance factors in mice in comparison with commercial formulation Ridostin.MATERIALS AND METHODS:dsRNA and Ridostin formulations were injected intramuscularly once at the dose of 5 mg/ml, polyglucinum--at the dose of 3.75 mg/ml. 3, 5, 24, 48 and 72 hours after the injection serum interferon levels, neutrophil oxidation-reduction activity parameters, peritoneal macrophage phagocyte activity levels were analyzed in mice blood samples.RESULTS:New dsRNA and polyglucinum containing composite formulation is a non-specific resistance system stimulator. dsRNA-PG effect on interferon synthesis and mice phagocyte activity was higher than with Ridostin and developed earlier. Neutrophil function activation by the formulation had a prolonged effect. A possible explanation for increased activity of dsRNA and polyglucinum composite formulation is a modulating effect by the polysaccharide component.CONCLUSION:The new formulation may have a more intensive and prolonged protective effect against influenza virus in comparison with Ridostin.

К числу перспективных противоопухолевых препаратов относятся представители семейства факторов некроза опухоли, в частности фактор некроза опухоли альфа (ФНО-α), который явля-ется противоопухолевым агентом, подавляя рост злокачественных новообразований и вызывая геморрагический некроз опухолей [5, 15]. В то же время результаты клинических испытаний препаратов рекомбинантного ФНО-α человека свидетельствуют о широком спектре его по-бочных эффектов. Этот факт, а также незна-УДК: 615.36:612.1]:616–006.04+57.084.1

Human tumor necrosis factor has a high antitumor activity. In this review we discuss the recent data presented by various countries on clinical trials of human tumor necrosis factor’ preparations.

Antitumor activity of TNF-α incorporated in nanoparticles (VLP-TNF-α) and dynamics of its accumulation and elimination from the blood and tumor tissue were studied in ICR mice. The VLP-TNF-α preparation exhibited higher antitumor activity compared to free TNF-α, presumably due to longer circulation of the cytokine in the blood and its more intensive accumulation by tumor tissue.

Human tumor necrosis factor has a high antitumor activity. In this review we discuss the recent data presented by various countries on clinical trials of human tumor necrosis factor’ preparations. Key words: tumor necrosis factor, oncology, clinical trials. Введение Поиск средств и способов лечения злокачест- венных новообразований продолжает оставаться актуальным направлением онкологии. Особое поло- жение занимают исследования, направленные на применение и совершенствование цитокинотерапии онкологических заболеваний. Фактор некроза опу- холи альфа (ФНО–α, ФНО, TNF) втечение многих лет привлекает внимание исследователей, фармако- логов, клиницистов, что связано с его избирательной способностью тормозить рост и вызывать лизис зло- качественных клеток, геморрагический некроз опу- холей, активировать иммунный противоопухолевый ответ, оказывать повреждающее действие на сосуды. Вместе с тем, результаты доклинических ис- следований и I–II фаз клинических испытаний ФНО-