Jennet Tumarovna Mammedova

Background: Ankylosing spondylitis (AS) associated with Crohn’s disease (CD) is a nosological form of spondyloarthropathies with a low population prevalence. The pathogenesis of this disease is not fully understood, and there are no precise differential diagnostic methods to distinguish AS associated with CD from AS and CD separately in the early stages of the disease. The main objective for this study is to define lymphocyte-mediated immunity in patients with AS associated with CD. Methods: For the pilot study, we recruited four groups: CD (n = 16), AS+CD (n = 13), AS (n = 13) and healthy controls (HC, n = 26). Immune phenotyping of peripheral blood lymphocytes was carried out via flow cytometry. Results: In the AS+CD group, circulating regulatory T-cell (Treg) levels were increased compared to the other groups. The frequency of CD73+ Tregs within the effector memory (EM) compartment was also elevated in the AS+CD group relative to the AS and CD groups alone. At the same time, the EM Tcyt and EM Th populations were lower in the AS+CD group compared to the CD group. Tfh17 levels were lower in the AS+CD group than in the AS group and showed a positive correlation with Bm5 and ‘switched’ memory B cells. Additionally, Tfh17 and ‘switched’ memory B cells correlated negatively with acute-phase markers, whereas Tfh2 correlated positively with the BASDAI activity index. Conclusions: In patients with AS associated with CD, exhaustion of the regulatory compartment of adaptive immunity is seen, whereas the humoral component appears oriented towards resolving the inflammatory responses.

Background: Ankylosing spondylitis (AS) associated with Crohn’s disease (CD) is a nosological form of spondyloarthropathies with a low population prevalence. The pathogenesis of this disease is not fully understood, and there are no precise differential diagnostic methods to distinguish AS associated with CD from AS and CD separately in the early stages of the disease. The main objective for this study is to define lymphocyte-mediated immunity in patients with AS associated with CD. Methods: For the pilot study, we recruited 4 groups: CD (n=16), AS+CD (n=13), AS (n=13) and healthy controls (HC, n=26). Immune phenotyping of peripheral blood lymphocytes via flow cytometry. Results: In the AS+CD group, circulating regulatory T cells (Treg) levels were increased compared to the other groups. The frequency of CD73+ Tregs within the effector memory (EM) compartment was also elevated in the AS+CD group relative to the AS and CD groups alone. At the same time, the EM Tcyt and EM Th populations were lower in the AS+CD group compared to the CD group. Tfh17 levels were lower in the AS+CD group than in the AS group and showed a positive correlation with Bm5 and switched memory B cells. Additionally, Tfh17 and switched memory B cells correlated negatively with acute-phase markers, whereas Tfh2 correlated positively with the BASDAI activity index. Conclusions: In patients with AS associated with CD, exhaustion of the regulatory compartment of adaptive immunity is seen, whereas the humoral component appears oriented towards resolving the inflammatory responses.

Multiple sclerosis (MS) is a chronic progressive neurodegenerative autoimmune disease characterized by disseminated demyelination patches in brain and spinal cord, containing different subsets of immune cells, including CD8+T cells. Currently, CD8+T cells may be subdivided into three main subsets, including Tc1, Tc2 and Tc17, according to their cytokine production profile and phenotype. A balance between the cytolytic Tc1 and cytokine-producing Tc2 and Tc17 cell subsets seems to play crucial role in emergence of diverse pathological conditions including autoimmunity. Thus, we have examined the frequency of Tc cell subsets in peripheral blood of patients with relapsing-remitting MS (MS, n = 25), and healthy individuals (HC, n = 24) matched for sex and age. To analyze the frequency of CD8+T cell subsets, we used multicolor flow cytometry. We evaluated the relative and absolute frequencies of Tc1 (CCR6-CXCR3+), Tc2 (CCR6-CXCR3-), Tc17 (CCR6+CXCR3-) and Tc17.1 (CCR6+CXCR3+) cells, like as their relative distribution along the main maturation stages of CD8+T cells, including ‘‘naïve’’ (CD45RA+CD62L+), central (СМ) and effector (ЕМ) memory, as well as TEMRA (CD45RA+CD62L-) cells. First, we have found that the relative frequency of Tc1 was decreased in MS group versus HC, whereas the relative and absolute frequencies of Тс17 of Тс17.1 were significantly elevated in MS patients. Next, our data revealed a significantly increased frequency of Тс17 cells at all analyzed stages of CD8+T cell maturation in peripheral blood samples from MS patients. Moreover, the differences against control group were more pronounced in the ЕМ and TEMRA CD8+T cell subsets which are able to migrate to inflammation sites (11.66% (4.75-14.69) versus 2.45% (1.48-3.89) and 4.91% (3.68-8.63) versus 0.41% (0.11-1.30), respectively, р 0.001 in both cases). Hence, we provide some new insights in the frequency of ‘‘polarized’’ CD8+T cell subsets in patients with MS. The obtained data suggest Tc17 cells to be an important part in MS pathogenesis which may be used for development of new diagnostic techniques and treatment approaches in MS patients.

Введение. CD4+ Т-лимфоциты играют ключевую роль в регуляции иммунитета и важны для противоопухолевой защиты. Они представляют собой гетерогенную популяцию, что требует избирательного анализа отдельных групп. Цель. Изучение дифференцировки и субпопуляционного состава CD4+ Т-лимфоцитов периферической крови у больных неоперабельной меланомой кожи по сравнению со здоровым контролем. Материалы и методы. В исследование включено 37 больных с диссеминированной или местнораспространенной нерезектабельной формой меланомы кожи. В группу сравнения включены здоровые доноры, соответствующие по возрастному и половому составу. С использованием многоцветной проточной цитометрии в периферической крови выявляли субпопуляции CD3+CD4+ лимфоцитов (Th): Th1, Th2, Th17, Tfh, а также клетки центральной (Th СМ) и периферической памяти (Th ЕМ). В рамках общего пула EM- и CM-клеток проводили детальный анализ отдельных субпопуляций Th17- («классических» Th17, Th17.1, DN Th17 и DP Th17) и Tfh-клеток (Tfh1, Tfh2, Tfh17 и DP Tfh). Результаты. При отсутствии количественных изменений в общем пуле CD4+ T-лимфоцитов крови у больных меланомой кожи выявлен дисбаланс отдельных субпопуляций, таких как Th1/Th2 среди CD4+ T-лимфоцитов, а также среди Th EM и среди Th CM. Кроме того, было повышено содержание клеток памяти Th CM и Th EM. Также выявлены сдвиги в балансе отдельных субпопуляций Th17 EM и Tfh EM. С тяжестью заболевания коррелировали такие показатели, как увеличение содержания Th17 и снижение Tfh CM-лимфоцитов, изменение соотношения субпопуляций клеток памяти (Th наивных и Th EM) и субпопуляций Th17 CM («классических» Th17 и Th17.1). В целом это указывает на наличие в организме больных активного иммунного ответа, как клеточного (изменения Th1, Th2 и Th17), так и гуморального (сдвиги в содержании Tfh). Выводы. Изучение отдельных субпопуляций CD4+ T-лимфоцитов периферической крови у больных меланомой более информативно, чем суммарная оценка большой гетерогенной группы клеток. Некоторые из выявленных показателей могут быть в дальнейшем использованы для оценки прогноза или эффективности проводимой терапии.

Hemostasis is a mechanism that stops bleeding from an injured vessel, involves multiple interlinked steps, culminating in the formation of a “clot” sealing the damaged area. Moreover, it has long been recognized that inflammation also provokes the activation of the coagulation system. However, there has been an increasing amount of evidence revealing the immune function of the hemostasis system. This review collects and analyzes the results of the experimental studies and data from clinical observations confirming the inflammatory function of hemostasis. Here, we summarize the latest knowledge of the pathways in immune system activation under the influence of coagulation factors. The data analyzed allow us to consider the components of hemostasis as receptors recognizing «foreign» or damaged «self» or/and as «self» damage signals that initiate and reinforce inflammation and affect the direction of the adaptive immune response. To sum up, the findings collected in the review allow us to classify the coagulation factors, such as Damage-Associated Molecular Patterns that break down the conventional concepts of the coagulation system.

BACKGROUND: The mTOR/S6K metabolic intracellular signaling cascade plays a key role in regulating the activation and differentiation of Th17/Treg lymphocyte populations. Pathogens can disrupt T-lymphocyte differentiation programs and weaken immune responses through arginine depletion mechanisms. AIM: To evaluate the effect of bacterial arginine-hydrolyzing enzyme arginine deiminase on the balance of Th17/Treg helper T-cell polarization in vitro. MATERIALS AND METHODS: Experiments were conducted using mononuclear leukocytes from healthy donors. Lymphocyte activation and differentiation were induced with activating antibodies and a cytokine cocktail. Recombinant Streptococcus pyogenes arginine deiminase was used. The enzyme’s arginine-hydrolyzing activity was confirmed by a modified Sakaguchi method. Mitochondrial oxidative phosphorylation activity was analyzed via MTT assay. The proportion of Th17, Treg, and IL-17A+/Foxp3+ transitional lymphocyte populations was assessed using monoclonal antibodies and flow cytometry. RESULTS: The enzyme caused a significant sevenfold decrease in arginine concentration in mononuclear leukocyte cultures. The arginine deficiency induced by arginine deiminase activity could not be compensated even by supplementing supraphysiological concentrations of the amino acid. Differentiation was accompanied by a significant reduction in oxidative phosphorylation (p 0.001). Arginine deiminase inhibited mitochondrial respiration in both intact (p 0.001) and differentiated cells (p 0.05). The enzyme’s activity suppressed Th17 cell differentiation (p 0.05) but did not affect IL-17A+/Foxp3+ transitional cells, Treg lymphocytes, or the Th17/Treg ratio. CONCLUSIONS: Arginine deiminase may disrupt adaptive immune development and diminish the effectiveness of protective immune responses.

The effectiveness of anticoagulant therapy in the prevention and treatment of pulmonary artery thrombosis and the possibility of anti-inflammatory therapy in preventing this complication in clinical practice and experiments were assessed. Data from patients with a new coronavirus infection and those suffering from urgent noninfectious pathology with confirmed pulmonary artery thrombosis were retrospectively analyzed. The outcomes of anticoagulant therapy and anticoagulant therapy combined with glucocorticoid and/or anticytokine drugs were assessed. Histological preparations of the lung vessels of patients were examined. Using an experimental model of rats with induced thrombosis of the posterior vena cava, changes in the pulmonary artery branches were assessed in the main group administered with edible mussel (Mytilus edulis) hydrolyzate and the control group given a physiological solution. No statistically significant relationship was found between the therapeutic, intermediate, and preventive anticoagulant therapy regimens and mortality, changes in lung dynamics, and D-dimer levels in 313 patients with new coronavirus infection. No predominance of any anticoagulant therapy regimen used was found among deceased patients. Thirty-nine patients were treated with glucocorticoid and/or anticytokine drugs in the presence of anticoagulant therapy. No statistically significant relationship in the onset of thrombotic complications was found between the groups receiving therapy with glucocorticoid and anticytokine drugs. No differences were noted in the drug-induced pathomorphosis of the wall of the pulmonary artery branches in the group receiving anticoagulant therapy or in the group receiving a combination of anticoagulant therapy and glucocorticoid and/or anticytokine drugs. Pulmonary artery thrombosis developed in all 19 patients suffering from urgent noninfectious pathology, 11 of whom were under anticoagulant therapy. In 12 of 15 rats in the control group with thrombosis of the posterior vena cava, blood clots were found in the lumen of the pulmonary artery branches. In 14 rats of the main group administered M. edulis hydrolyzate, no blood clots were found in the pulmonary artery branches. Thus, the systemic effects of anticoagulant therapy were offset by the local prothrombotic effects of the vascular wall caused by inflammation. Glucocorticoid and anticytokine drugs did not affect inflammatory changes in the vascular wall and did not prevent pulmonary artery thrombosis. The introduction of M. edulis in the experiment prevented pulmonary artery thrombosis in the presence of posterior vena cava thrombosis, which indicates a promising direction in the search for pathogenetic prevention of this complication.

Pathogenic microbes use arginine-metabolizing enzymes as an immune evasion strategy. In this study, the impact of streptococcal arginine deiminase (ADI) on the human peripheral blood T lymphocytes function in vitro was studied. The comparison of the effects of parental strain (Streptococcus pyogenes M49-16) with wild type of ArcA gene and its isogenic mutant with inactivated ArcA gene (Streptococcus pyogenes M49-16delArcA) was carried out. It was found that ADI in parental strain SDSC composition resulted in a fivefold decrease in the arginine concentration in human peripheral blood mononuclear cell (PBMC) supernatants. Only parental strain SDSCs suppressed anti-CD2/CD3/CD28-bead-stimulated mitochondrial dehydrogenase activity and caused a twofold decrease in IL-2 production in PBMC. Flow cytometry analysis revealed that ADI decreased the percentage of CM (central memory) and increased the proportion of TEMRA (terminally differentiated effector memory) of CD4+ and CD8+ T cells subsets. Enzyme activity inhibited the proliferation of all CD8+ T cell subsets as well as CM, EM (effector memory), and TEMRA CD4+ T cells. One of the prominent ADI effects was the inhibition of autophagy processes in CD8+ CM and EM as well as CD4+ CM, EM, and TEMRA T cell subsets. The data obtained confirm arginine’s crucial role in controlling immune reactions and suggest that streptococcal ADI may downregulate adaptive immunity and immunological memory.

Тромбоз — обтурация венозных или артериальных кровеносных сосудов, которая в ряде случаев может приводить к фатальному исходу. Разработка эффективных препаратов, направленных на терапию этого состояния, остается актуальной научно-практической задачей. ЦЕЛЬ ИССЛЕДОВАНИЯ Изучить антитромботические эффекты препарата кислотного гидролизата голубых мидий (Mytilus edulis). МАТЕРИАЛ И МЕТОДЫ С помощью биохимических методов исследовали влияние гидролизата M. edulis на ферментативную активность ключевых сериновых протеаз каскада коагуляции. Для анализа влияния препарата на биологические эффекты тромбина изучали структуру актинового цитоскелета и морфологию монослоя эндотелия in vitro. Антикоагулянтное действие препарата исследовали в модели тромбоза задней полой вены у крыс in vivo. РЕЗУЛЬТАТЫ Было обнаружено, что препарат ингибирует ферментативную активность протеаз каскада свертывания (FIIa, FXa, FXIa, FXIIa), в частности ферментативную активность тромбина (FIIa) в отношении фибриногена и синтетических субстратов Tos-Gly-Pro-Arg-pNA и Ala-Ala-Arg-pNA. Антитромбиновая активность неразбавленного препарата составляла около 6,5 АТ-U/мл. Препарат подавлял способность тромбина вызывать ремоделирование актинового цитоскелета и дезинтеграцию монослоя эндотелия. Подкожные инъекции препарата приводили к значительному изменению структуры венозного тромба и снижению вероятности его образования в модели тромбоза задней полой вены у крыс. ЗАКЛЮЧЕНИЕ Гидролизат M. edulis может служить основой для разработки новых препаратов для терапии состояний, связанных с гиперкоагуляцией.

More than a century has passed since arginine was discovered, but the metabolism of the amino acid never ceases to amaze researchers. Being a conditionally essential amino acid, arginine performs many important homeostatic functions in the body; it is involved in the regulation of the cardiovascular system and regeneration processes. In recent years, more and more facts have been accumulating that demonstrate a close relationship between arginine metabolic pathways and immune responses. This opens new opportunities for the development of original ways to treat diseases associated with suppressed or increased activity of the immune system. In this review, we analyze the literature describing the role of arginine metabolism in the immunopathogenesis of a wide range of diseases, and discuss arginine-dependent processes as a possible target for therapeutic approaches.