Objective. The work was performed with the purpose to study microbiota state of the gastro-intestinal tract (GIT) of C57Bl/6 mice with signs of metabolic dysbiosis (MD) induced with the use of fructose solution or with poloxamer 407 (P 407), during treatment with combination of sodium polyprenylphosphate (PPP) and beta-sitosterol (BSS). Materials and Methods. MD was induced in mice with drinking 20 per cent water fructose solution. MD was also induced in mice by intraperitoneal use of P 407 at the dose of 500 mg/kg two times per week during ten weeks. The mice of experimental groups received intragastrically combination of PPP and BSS preparations (0,2 ml of the suspension per mouse). Medical scheme consisted in use of preparations five times per week starting after one month from the first day of fructose drinking or of P 407 use and prolonging for one month else. The quantity of viable bacteria in the probes obtained was measured by the direct passage of the material tested on elective or selective nutritional media with following count of colonies grown. Results. The drink of fructose solution or use of P 407 caused the changes of GIT microbiota state that confirmed development of signs of MD in mice. Treatment with PPP and BSS combination was shown to cause actual tendencies to restore the normal GIT microbiota state. Conclusion. Combination of PPP and BSS preparations seemed to be efficient as a novel approach to correction of GIT microbiota state perturbed due to pathogenesis of MD considered as one of the primary signs of metabolic syndrome in mice.

The aim of the research was to study the population composition of the splenic lymphoid cells, to assess the functional activity of lymphocytes as well as the state of the gastrointestinal tract microbiota in experimental modeling of metabolic syndrome (MS).The studies were conducted using two experimental models of MS and hyperlipidemia (HL), based on prolonged drinking of animals with 20% aqueous fructose solution with added cholesterol and intraperitoneal administration of Poloxamer 407 to mice, respectively.The results of the experiments indicate a change in the population composition of splenocytes (decrease in CD4+ and CD8+T cells, activation of CD4+CD25+FoxP3+Thed cells), accompanied by a decrease in T cell activity and increased proliferation of B lymphocytes, impaired production of IL-15 and IL-22, as well as lipid and carbohydrate metabolism (adiponectin, leptin, insulin), which serves as a prerequisite for the development of chronic inflammation, which is a pathogenetic sign of MS.We found changes in the intestinal microbiota of mice characteristic of the manifestation of metabolic dysbiosis – an increase in the representation of Firmicutes bacteria (staphylococci, streptococci, enterococci) in the biomaterial, changes in the content of facultative (E. coli) and transient (Enterobacter) microflora.In order to develop a new kind of medicine for therapy and prevention of HL and MS, we used a combination of sodium polyprenyl phosphate (PP) and beta-sitosterol (BSS), polyisoprenoid derivatives of plant origin.More pronounced changes were found in the splenocyte population composition and activation parameters of Treg cells in HL modeling compared with the MS model. The introduction of PP and BSS has an immunocorrective effect during treatment.The therapeutic effect of this drug, as well as the prevention of the MS symptoms, is accompanied by normalization of the microbiota state.The data obtained indicate the prospects of using PP and BSS for the prevention and treatment of HL and MS in order to influence the leading links in the pathogenesis of metabolic disease.

Helicobacter pylori (HP) bacteria have a wide range of pathogenicity factors. The HP genome contains genes of the CagA group (cytotoxin-associated genes). Infection with CagA-positive HP strains is associated with the production of pro-inflammatory cytokines, which are involved in the maintenance and development of destructive and inflammatory changes in the organism. Here we studied the role of CagA proteins in the regulation of the immune response in DBA mice and evaluated the corrective effect of polyprenyl phosphate (PP) on this process. Genetically modified strains of E. coli were used, differing by the presence of an island of genes encoding the synthesis of HP CagA proteins. The drug Phosprenyl was used as a source of PP. The subpopulation composition of spleen cells was evaluated by flow cytofluorimetry using monoclonal antibodies. The level of IL 10, TGF-β in blood serum was determined by the enzyme immunoassay. The proliferative activity of splenocytes was measured by the standard procedure based on the inclusion of 3 H-thymidine in the DNA. CagA HP proteins caused the polarization of the immune response by the Th1-dominant type, which was expressed by an increase in the population of CD4 + cells, CD25 + and CD25 + Foxp3 + T cells and increase proliferation T- limphocytes. Inoculation of CagA + bacteria was accompanied by a quantitative increase in TGF-β produced by activated Treg cells. PP inoculation led to the normalization of the CD4 and CD8 T cells, a decrease in the populations of CD25 + , Foxp3 + , CD25 + Foxp3 + cells, and further increase in the IL-10 levels. Thus, PP corrected cellular immune response, prevented the pronounced polarization of the Th1 immune response, and reduced the activation of Treg population. The results obtained indicate a possible decrease in the pro-inflammatory immune response under the influence of PP.

Metabolic syndrome (MS) is a widespread polyethiological clustering characterized by metabolic, hormonal disorders and clinical manifestations that significantly increase the risk of developing cardiovascular diseases, atherosclerosis, type II diabetes and other pathological conditions. An important role in the development of MS is assigned to the intestinal microbiota. To develop new therapeutic agents for correction of MS manifestations, it is necessary to develop adequate experimental models. In this paper, comparative studies were conducted to assess the parameters of lipid metabolism, the content of peptide hormones, morphological changes in liver tissue, and the quantitative and generic composition of the intestinal microbiota of mice. Experimental models of experimental hyperlipidemia (HL) caused by the introduction of poloxamer 407 (Pol407) and alimentary MS (a diet with fructose and the addition of cholesterol to the feed) were used. Significant increase in the levels of cholesterol, triglycerides, and low-density lipoprotein (LDL) was found in the group of mice treated with Pol407 injections. To assess the indicators of carbohydrate metabolism in blood serum, the following markers were determined: insulin, adiponectin, leptin. In the alimentary MS model a decrease in adiponectin in the blood serum, while insulin level was increased. In both experimental models, significant changes in the gut microbiota of mice were observed. They were associated with the manifestation of metabolic dysbiosis - an increase in the representation of Firmicutes (staphylococci, streptococci, enterococci) in the biomaterial, changes among representatives of both facultative (E. coli), and transient (Enterobacter bacteria) microflora. In addition, dystrophic, as well as morphological changes and signs of inflammation in the liver tissue were noted in both groups.

Изучение иммунного ответа и состояния метаболизма на экспериментальных моделях развития гиперлипидемии и метаболического синдромаФедеральное государственное бюджетное учреждение «Национальный исследовательский центр эпидемиологии и микробиологии имени почетного академика Н.Ф.Гамалеи» Министерства здравоохранения Российской Федерации, 123098, г.Москва, Российская Федерация Резюме Введение.Иммунная система выполняет важную роль в патогенезе гиперлипидемии (ГЛ) и метаболического синдрома (МС).Метаболические дисфункции сопровождаются нарушениями кооперации, межклеточного взаимодействия различных звеньев иммунной системы.К ключевым патогенетическим факторам ГЛ и МС относятся изменение секреции медиаторов адипоцитов (адипонектина, лептина), провоспалительных и противовоспалительных цитокинов (ЦТ).Данные экспериментальных и клинических исследований свидетельствуют о взаимосвязи и взаимообусловленности клеточных иммунных реакций, медиаторов межклеточных взаимодействий и метаболических нарушений макроорганизма.Цель исследования -изучение популяционного состава лимфоидных клеток селезенки, продукции цитокинов и функциональной активности лимфоцитов при экспериментальном моделировании МС и ГЛ.Материал и методы.Эксперименты выполнены на мышах C57Bl/6.Исследования проведены на 2 экспериментальных моделях МС и ГЛ, основанных на длительном выпаивании животных 20 % водным раствором фруктозы с добавлением в корм холестерина и внутрибрюшинном введении мышам Полоксамера 407 соответственно.Оценка субпопуляционного состава клеток селезенки мышей проведена методом проточной цитометрии.Определение лептина, адипонектина, инсулина, цитокинов интерлейкина(ИЛ)-15, ИЛ-22 в сыворотке крови мышей проводили методом ELISA с использованием коммерческих наборов.Пролиферацию лимфоцитов оценивали в реакции бласттрансформации (РБТЛ).Результаты экспериментов при моделировании МС и ГЛ свидетельствуют об изменении популяционного состава селезенки (снижение содержания CD4 + -и CD8 + -Т-клеток, активация CD4 + CD25 + Foxp3 + -Тreg-клеток), сопровождающимся снижением функциональной активности иммунокомпетентных Т-клеток и увеличением пролиферативной активности В-лимфоцитов, нарушением продукции ИЛ-15, -22, участвующих в липидном обмене, и о дисфункциональных изменениях медиаторов липидного и углеводного обмена (адипонектин, лептин, инсулин), что в совокупности является предпосылками для развития хронического воспаления -важнейшего патогенетического признака МС.Заключение.Полученные данные по изменению иммунного ответа и состояния метаболизма подтверждают целесообразность поиска и скрининга иммунокорригирующих лекарственных средств, предназначенных для профилактики и лечения гиперлипидемии и симптомокомплекса МС, исходя из целенаправленного воздействия на ведущие звенья патогенеза метаболической болезни.

Feline enteropathogenic coronavirus affects the intestinal epithelium and causes inflammation of the intestinal mucosa in cats. In adult cats, the infection is usually asymptomatic, but the constant circulation of the coronavirus in the feline population can increase the likelihood of mutation leading to appearance of the virulent strain of the virus that cause feline infectious peritonitis - a deadly disease that develops in 5-10% of cats carrying the coronavirus. The aim of this work was to study the effect of the drugs Phosprenyl and Gamavit on the course of coronavirus infection in cats. A total of 29 cats were monitored. The diagnosis was confirmed by the detection of antibodies by the IHA method, as well as by ultrasound examination of the abdominal organs. Prescribed treatment: Phosprenyl per os, 1 ml , twice a day, course 2-4 months; Gamavit 1 time a day, s/c, course 10 days, with a repeated course after 3 weeks; 3-4 times. As symptomatic therapy, drugs were used to improve the condition of the stool. At the end of treatment, symptoms of gastrointestinal tract damage disappeared in all patients. If before the start of treatment, most patients showed signs of anemia (a decrease in the number of red blood cells and hemoglobin levels), leukocytosis with a predominance of segmented neutrophils, characteristic of infectious and inflammatory processes, and increased ESR, then at the end of therapy, these indicators were completely normalized. Thus, long-term therapy with the use of Phosprenyl and Gamavit helps to control the course of coronavirus infection in cats, prevent its exacerbation and transition to a highly lethal form of infectious peritonitis.

The immunomodulatory properties of immunobiological drugs Glutoxim and Phosprenyl we well as vesicular stomatitis virus and inactivated tick-borne encephalitis vaccine virus were studied using human diploid fibroblast cell line from the collection of M. P. Chumakov Federal Research Center for Research and Development of Immunobiological Products. All tested preparations exhibited immunomodulatory activity in human diploid fibroblast cell line. Glutoxim in doses of 0.1 and 0.25 μg/ml stimulated production of IL-6 and IL-10 during 24-48 h of culturing, but did not stimulate production of IL-1β. Phosprenyl, on the contrary, increased production of IL-1β and the levels of IL-6 and IL-10. Vesicular stomatitis virus stimulated the production of IL-1β, IL-6, and IL-10, while inactivated tick-borne encephalitis vaccine virus stimulated the production of cytokines IL-8 and IL-18. Immunomodulatory activity of inactivated tick-borne encephalitis vaccine virus was first demonstrated in the in vitro system.

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Antiapoptosis activity of plant polyprenylphosphate against macrophage target cells infected with the murine encephalomyelitis virus. Infection caused by the Theiler’s murine encephalomyelitis virus (TMEV) is regarded as an experimental model of multiple sclerosis, since both of these diseases are characterized by similar pathology of the central nervous system tissues and involvement of the immune system in the development of the demielinization. The aim of the work was to study the effect of plant-derived polyprenylphosphate (PP) on the apoptosis of infected target cells. We showed that PP reduced apoptosis of macrophage target cells infected with TMEV. It is known that in the protocol of multiple sclerosis treatment some medicines possessing immunomodulatory, antiviral, anti-inflammatory and antioxidant activity are used. Since PPs of plant origin also have all these activities, the prospects of their use as therapeutic agents are discussed.

We studied the sensitivity of domestic proprietary human and animal cell lines from the collection of M. P. Chumakov Federal Scientific Center for Research and Development of Immuneand-Biological Products to infection with different enterovirus 71 strains. A cell system based on domestic proprietary permanent cell line 4647 was for the first time used for reproduction of four enterovirus 71 strains (BrCr, 42266, 42934, and 43374). It was shown that strain 4647 is the optimal cell substrate for enterovirus 71 reproduction. The titers of enterovirus 71 for all four strains considerably (by 2 lgTCID 50 /ml and more) increased during sequential passages in permanent cell line 4647. The prospects of using permanent cell line 4647 for creation of diagnostic and preventive preparations against 71 was demonstrated.

Рассмотрена роль первичных фибробластов и линий диплоидных клеток фибробластов человека в формировании противоинфекционного иммунитета. Обсуждается адекватность клеточной модели на основе фибробластов для изучения молекулярных механизмов иммунопатогенеза вирусных инфекций. Рассмотрена возможность использования фибробластов для изучения особенностей действия иммунотропных препаратов. Обсуждается перспективность клеточной модели на основе аттестованных линий диплоидных клеток фибробластов для изучения механизмов действия новых и известных иммуномодуляторов.