The aim – to evaluate subpopulations of B lymphocytes and features of interferon (IFN) status in patients with systemic lupus erythematosus (SLE), to clarify the relationship of immunological parameters with clinical manifestations of the diseaseMaterial and methods. 139 patients (123 (88%) women and 16 (12%) men) with a definite diagnosis of SLE were included in the analysis. The disease duration was 3.0 [0.3; 12.0] years, SLEDAI-2K (Systemic Lupus Erythematosus Disease Activity Index 2000) – 7 [4; 11] points, SDI (Systemic Lupus International Collaborating Clinics Damage Index) – 0 [0; 1] points. Immunophenotyping of peripheral blood lymphocytes, including determination of B cells, the general population of memory B cells, non-switched and switched memory B cells, naive, transient B cells, and plasmablasts was carried out using multicolor flow cytometry. IFN status was assessed by the expression of IFN-stimulated genes (MX1, RSAD2, EPSTI1) using real-time polymerase chain reactionResults. Two immunological “patterns” were identified – the prevailing immunological mechanism of the pathogenesis of the disease – SLE – with predominant activation of type I IFN and with predominant activation of the B cell component of the immune system. The immunological phenotype with activation of type I IFN was associated with high immunological activity, predominant skin damage, leukopenia, and the phenotype with predominant activation of the B cell link was associated with damage to the kidneys and nervous system.Conclusion. The results of the work suggest a wide variety of immune mechanisms underlying the pathogenesis of SLE. It is possible to identify a number of leading molecular “patterns” of the pathogenesis of the disease, which must be taken into account to select an effective “targeted” drug.

The growth of malignant tumors is accompanied by an increase in plasmin activity, one of the key enzymes in oncogenesis. IgG proteolysis by plasmin yields fragments with exposed C-terminal lysine residues that can bind to plasmin heavy chain. Highly purified plasminogen (plasmin precursor) and heavy chains containing lysine-binding sites were isolated. The formation of IgG fragments as a result of their proteolysis by plasmin in solution was shown. Using ELISA with immobilized heavy chains, we found increased concentration of IgG fragments in the sera of patients with esophageal cancer in comparison with healthy donors. The test had high sensitivity (91%) and specificity (85%).

We studied the effect of conditioned media (CM) from cultivated bone marrow stromal cells grown at 10% O2 on extracellular matrix and vascular component in the healing zone after surgical incision of the uterine wall in Sprague-Dawley rats (n=17). Control group (n=10) received no treatment. On days 5 and 15 after the surgery, the expression of CoL1a1, CoL4a, MMP9, TIMP1, and FGF2 genes was evaluated and a morphological study was carried out. On day 5, CoL1a1 expression, CD34+ cell content, and the area of newly formed tissue were lower in the experimental group. On day 15, the expression of MMP9, TIMP1, FGF2, and CoL1a1 genes in the control group was lower, while CD34+ cell content and area of healing zone were higher. Thus, the application of CM reduced the damage area and accelerated the recovery process after surgical full-thickness incision of the uterine wall.

Introduction. Placenta accreta spectrum is an abnormal invasion of villous trophoblast into the myometrium and one of the most severe complications of pregnancy. We aimed to study morphofunctional, molecular, and immunological changes in the placenta and peripheral blood in various types of placenta accreta spectrum.Materials and methods. The study involved 45 pregnant women who underwent ultrasound examination at weeks 35–38. According to the ultrasound data, 15 women had placenta accreta and 15 women developed placenta increta. The comparison group included 15 pregnant women without placenta accreta spectrum with a uterine scar after cesarean section. Histological and immunohistochemical studies were performed on paraffin sections of the placentas. We used mouse antibodies to insulin-like growth factor-binding protein-1, pregnancy-specific β-1 glycoprotein, and human placental lactogen, proteinase 3 and measured them using ELISA. We also studied tumor necrosis factor. Immunophenotyping of peripheral blood lymphocytes was performed using flow cytometry. Results. Рlacenta accreta spectrum is accompanied by increased intensity of IHC staining of placental villi for all the studied markers, in contrast to those in the comparison group. The maximum intensity of insulin-like growth factor-binding protein-1 staining was found in placenta increta. Increased serum concentration of proteinase 3 and placental lactogen (placenta increta) and decreased level of insulin-like growth factor-binding protein-1 were found in placenta accreta spectrum.Conclusion. In placenta accreta spectrum, there is a functional redundancy of placental proteins and an increased pro-inflammatory status. An imbalance between aggressive factors are likely to contribute to pathological villous invasion. Keywords: placenta accreta spectrum, immune status, placental proteins, proteinase 3

The aim of our study was to assess the relationship between the changes of antinuclear autoantibodies (ANA) and autoantibodies to topoisomerase 1 (anti-Topo 1) in systemic sclerosis (SSc) patients on rituximab (RTX) therapy.Materials and methods. The prospective study included 88 patients (73 women) with a mean age of 47 (17– 71) years. The mean disease duration was 5.9±4.8 years. The mean follow-up period was more than 2 years (27 (12–42) months).Results. We documented a statistically significant change in skin score, the disease activity index, improvement of pulmonary function and reduction of mean dose of prednisolone after RTX treatment. There was a significant decrease in the number of patients with high levels of ANA and overall decrease of the ANA and anti-Topo 1 levels. A moderate positive statistically significant correlation was found between ANA and anti-Topo 1 (r=0.403). In the group of patients positive for anti-Topo 1 there were a more pronounced depletion of B lymphocytes, significantly higher increase in forced vital capacity and diffusion capacity, decrease in the disease activity index, compared with a patients negative for anti-Topo 1.Conclusions. We observed the decline in the level of ANA and anti-Topo 1 in SSc patients after RTX therapy and it was correlated by an improvement of the main outcome parameters of the disease. Therefore, anti-Topo 1 positivity could be considered as a predictor of a better response to RTX treatment, especially in SSc patients with hyperproduction of anti-Topo 1.

Blood serum of patients with gastric (n = 68) and esophageal (n = 43) cancer was assessed for proteolytic fragments of IgG. Serum samples of 20 healthy donors were used as a control. We analyzed indicators of hemostasis (prothrombin time, fibrinogen, plasminogen activity, a2-antiplasmin activity, protein C activity) in blood plasma and the level of total IgG in the blood serum. The median IgG-LysK of healthy donors was lower than in esophageal cancer and in patients with gastric cancer. ROC-analysis showed high sensitivity (91%) and specificity (85%) in the group with esophageal cancer but 68% and 85%, respectively, in patients with gastric cancer. Analysis of false negatives IgG-LysK in cancer patients showed that most patients had an advanced stage of cancer accompanied by metastases. Total IgG in the plasma of patients with false-negative IgG-LysK values was 30% lower than in samples with positive values, while the level of a2-antiplasmin was increased and the prothrombin time was shorter. These changes in blood homeostasis may be the reason for an increase in the proportion of false-negative values of the IgG-LysK coefficient. Circulatory IgG-LysK levels increase in the early stages of such cancers as gastric and esophageal cancers. Thus, when used in a panel with other more specific markers for these pathologies, this indicator can significantly increase the early detection of cancer.

Uncontrolled activation of neutrophils is considered an important mechanism of thromboinflammation and fibrosis in immunemediated rheumatic diseases (IMRD), malignant neoplasms, atherosclerosis, COVID-19 and many other acute and chronic inflammatory diseases of humans. Particular attention has been drawn to the ability of neutrophils to form “network” (web-like) structures, called “neutrophil extracellular traps” NETs. The process associated with the formation of NETs and the weakening of their degradation is called “NETosis”. The publication summarizes data on the role of NETosis in the pathogenesis of IMRD and discusses the prospects for pharmacotherapy aimed at preventing the formation and destruction of NETs.

Objective : To study the frequency of spontaneous foci of DNA double-strand breaks (DSBs) in patients with immune-inflammatory rheumatic diseases (IIRD), their relationship to disease activity, levels of inflammatory markers, and levels of autoantibodies. Material and methods . The analysis included 40 patients with IIRD, including 19 patients with rheumatoid arthritis (RA, including 16 women, median disease duration 60 [20; 103] months, DAS28 was 5.05 [4.06; 5.9]) and 21 patients with systemic lupus erythematosus (SLE, 19 women, median disease duration 96.0 [40.0; 158.0] months, SLEDAI-2K 8.0 [4.0; 12.0]). The control group consisted of 17 healthy donors matched for sex and age. DNA DSBs were identified as discrete foci by immunofluorescence staining of lymphocyte cultures with antibodies against γH2AX and 53BP1 and subsequently analysed using the automated AKLIDES automated platform (Medipan). Results and discussion . There were no significant differences in the number of spontaneous DNA DSBs in patients with RA and healthy donors (p>0.05), a lower number of cells with the 53BP1 focus and a lower percentage of cells damaged in this focus were found in patients with SLE than in controls. There was a positive correlation between the number of γH2AXdamaged cells and CDAI(r=0.45, p=0.035), the number of cells with 53BP1 ruptures and the level of rheumatoid factor IgM (r=0.63, p=0.005) and ESR (r=0.53, p=0.02). In the group of SLE patients, a positive correlation was observed between the number of cells with breaks in the γH2AX focus and the level of antibodies against double-stranded DNA (anti-dsDNA; r=0.56, p=0.007), the average number of breaks in the cell in the γH2AX focus with the level of anti-dsDNA (r=0.57, p=0.004). Conclusion . The number of DNA DSBs may be an additional indicator of IIRD activity. In patients with SLE, DNA repair processes appear to be impaired, which is associated with the high activity of the disease.

We have suggested that adipocytes in uterine scars may affect the development of the placenta accrete spectrum (PAS). In the experimental part, we explored adipocytes in the uterine wall by the twelfth sexual cycle after surgery. In the clinical part, we investigated adipocyte clusters in the cesarean scar of pregnant women with and without PAS. The uterine wall was evaluated in gross and histological sections using morphometry, histochemistry (hematoxylin and eosin stain, Mallory stain), and immunohistochemistry for FABP4 (adipocyte markers), CD68, CD163, CD206 (macrophages), CD 34 (endothelium), cytokeratin 8 (epithelium), aSMA (smooth muscle cells). The design included an experimental study on Sprague–Dawley rats (n = 18) after a full-thickness surgical incision on the seventh (n = 6), 30th (n = 6), and 60th day (n = 6). The clinical groups include pregnant women without uterine scars (n = 10), pregnant women with a uterine scar after previous cesarean sections (n = 10), and women with PAS (n = 11). Statistical processing was carried out using nonparametric methods. Comparisons were conducted using the Mann–Whitney U-test and Kruskal–Wallis test. Statistical significance was considered at p < 0.05. On the seventh day, the rat uterine horn was enveloped by adipose tissue, which contained crown-like structures with FABP4+, CD68+, CD206+, and CD163+ cells. FABP4+ cells in the uterine wall were absent by the 30th day. The number of CD206+ and CD163+ cells in the adipose tissue decreased by the 30th day. On the 60th day, the attachment of fat tissue was revealed in the form of single strands. The serous layer around the damaged area totally recovered on the 60th day. FABP4+ cells were not detected in the uterine wall samples from pregnant women without a previous cesarean section. Adipocytes were found in the scar during non-complicated pregnancy and with PAS. Reducing the number of CD68+ cells in adipocyte clusters, there were in myometrium with PAS. Increased CD206+ and CD163+ cells were revealed in uterine adipocyte clusters of the group. According to the experimental finding, adipocytes should be absent in the uterine wall by the 12th sexual cycle after a full-thickness surgical incision. The presence of adipocyte clusters in cesarean scar indicated the disturbance of cell interaction. Differences in the numbers of CD206 and CD163 cells in adipocyte clusters between groups with and without PAS may be indirect evidence that uterine adipocytes affect the development of PAS.

ЦЕЛЬ ИССЛЕДОВАНИЯ Изучить показатели трансформирующего фактора роста бета-1 (ТФФ бета-1), фактора некроза опухоли альфа (ФНО альфа) и гликоделина A (ГдА) в перитонеальной жидкости, эндометриоидных инфильтратах и биоптатах тазовой брюшины при впервые выявленном и рецидивирующем наружном генитальном эндометриозе. МАТЕРИАЛ И МЕТОДЫ Обследованы 48 больных с рецидивирующим наружным генитальным эндометриозом (основная группа). В группу сравнения были включены 40 пациенток с впервые выявленным наружным генитальным эндометриозом. Контрольную группу составили 10 женщин без клинико-лабораторных проявлений эндометриоза. У всех больных в перитонеальной жидкости были исследованы уровни ГдА и ТФР бета-1 методом иммуноферментного анализа. В тканях эндометрия определяли уровни экспрессии генов ТФР бета-1 и ФНО альфа на основании измерения количества мРНК интересующего гена методом полимеразной цепной реакции (ПЦР) в режиме реального времени. РЕЗУЛЬТАТЫ Полученные результаты продемонстрировали, что уровни ТФР бета-1 в перитонеальной жидкости при рецидивирующем и впервые выявленном наружном генитальном эндометриозе были достоверно выше, чем у пациенток контрольной группы. Исследование ГдА в перитонеальной жидкости выявило в 20% наблюдений группы сравнения и 30,1% наблюдений основной группы чрезвычайно высокие его значения, превышающие в десятки раз максимальные показатели в контрольной группе. В интактной тазовой брюшине выявили повышенную экспрессию гена ТФР бета-1 при эндометриозе по сравнению с этим показателем в контрольной группе, максимальную при рецидивирующем наружном генитальном эндометриозе в основной группе. Экспрессия гена ФНО альфа в эндометриоидном инфильтрате была более выраженной при впервые выявленном наружном генитальном эндометриозе по сравнению с рецидивирующим. В интактной тазовой брюшине уровни гена ФНО альфа при наружном генитальном эндометриозе были выше, чем в контрольной группе. ЗАКЛЮЧЕНИЕ Таким образом, высокие уровни ГдА при наружном генитальном эндометриозе (20% наблюдений группы сравнения и 30,1% наблюдений основной группы), превышающие в десятки раз максимальные показатели контрольной группы, свидетельствуют, что гликоделин может стать биомаркером для диагностики и прогнозирования наружного генитального эндометриоза, а учитывая его иммунорегуляторные свойства, можно предполагать, что гликоделин является возможной мишенью для иммунотерапии.

Objective: assessment of the dynamics of T- and B-lymphocytes subpopulations in rheumatoid arthritis (RA) during therapy with synthetic disease-modifying antirheumatic drugs (sDMARDs) and biological disease-modifying antirheumatic drugs (bDMARDs): inhibitors of tumor necrosis factor α (iTNFα) and an inhibitor of interleukin 6 receptors (iIL6R ).Patients and methods. The study included 77 patients with RA who met the 2010 ACR/EULAR criteria (mean age 56 [44; 62] years). Group 1 included 30 (27 women and 3 men) patients with early RA who had not previously received therapy. Group 2 included 20 (14 women and 6 men) patients on sDMARD therapy who were prescribed iTNFα for the first time. The 3rd group is represented by retrospective data of 27 (23 women and 4 men) patients who previously used sDMARDs (MT – 85%, leflunomide – LEF – 15%), in whom iIL6R therapy was initiated for the first time. All study participants initially and 6 months later underwent immunophenotyping of T- and B-lymphocytes by flow cytofluorometry according to the standard method.Results and discussion. In all groups, there were no significant changes in the studied T-lymphocyte profile during 6 months of follow-up. When comparing the immunogram data of patients treated with sDMARDs and iTNFα, no significant differences in subpopulations of B-lymphocytes were found. At baseline, the iIL6R group had higher levels of naive B-lymphocytes and plasmablasts and low concentrations of «switched» B-cells. For all methods of treatment, the number of «switched» B-cells decreased, while plasmablasts and plasma cells increased.Conclusion. From the data obtained, it follows that the simultaneous decrease in the levels of memory B-cells and their «switched» forms, plasmablasts and plasma cells can be used as a marker for the early administration of drugs that disrupt the differentiation of B-lymphocytes, in particular, iIL6R.

(1) Background: The components of the fibrinolytic system and its main component, plasminogen, play a key role in the first months of pregnancy. The effect of autoantibodies interacting with plasminogen in the formation of retrochorial hematoma is unknown. The aim of our study was to determine the role of plasminogen and IgA, IgM, and IgG, which bind to plasminogen, in retrochorial hematoma. (2) Methods: Prothrombin time (PT), thrombin time (TT), partial activated thromboplastin time (aPTT), soluble fibrin-monomer complex (SFMC), D-dimer, plasminogen activity (%Plg), plasminogen concentration (Plg), and the levels of IgG (IgG-Plg), IgM (IgM-Plg), IgA (IgA-Plg) interacting with plasminogen were determined in plasma samples of 57 women with normal pregnancy and 16 with retrochorial hematoma. (3) Results: %Plg in plasma samples from women with retrochorial hematoma was significantly lower than in plasma samples from women with normal pregnancy. The diagnostic significance of %Plg in the ROC analysis was AUC = 0.85. A direct correlation was found between aPTT and the level of autologous IgM interacting with plasminogen. (4) Conclusions: A decrease in the activity of plasminogen in the blood serum of women in the first trimester of pregnancy may indicate disturbances in the hemostasis system and the formation of retrochorial hematoma. According to the results of the study, it is possible to recommend the determination of plasminogen activity in the management of pregnant women in gynecological practice.

ЦЕЛЬ ИССЛЕДОВАНИЯ Изучить диагностическую значимость уровней трансформирующего фактора роста бета-1 (ТФР-β1), фактора некроза опухоли альфа (ФНО-α) и гликоделина A (ГдА) при патологии эндометрия в пре- и постменопаузе. МАТЕРИАЛ И МЕТОДЫ Обследованы 120 женщин с различной патологией эндометрия: 73 пациентки в пременопаузе (1-я основная группа) и 47 пациенток в постменопаузе (2-я основная группа). Контрольные группы составили женщины, находящиеся в пременопаузе (n=10) и в постменопаузе (n=10), у которых отсутствовала патология эндометрия. У всех пациенток методом иммуноферментного анализа были определены уровни ГдА и ТФР-β1 в сыворотке крови. Кроме того, у всех пациенток проведено определение уровня экспрессии ТФР-β1 и ФНО-α, основанное на измерении количества матричной рибонуклеиновой кислоты интересующего гена в тканях эндометрия методом полимеразной цепной реакции в режиме реального времени. РЕЗУЛЬТАТЫ Уровни ТФР-β1 в сыворотке крови пациенток с патологией эндометрия в пременопаузе были статистически значимо ниже, чем в контрольной группе. Выявили статистически значимо более высокие уровни гликоделина A у женщин в пременопаузе по сравнению с этими показателями в группе постменопаузы. Экспрессия генов ФНО-α была более высокой у пациенток с патологией эндометрия в постменопаузе. ЗАКЛЮЧЕНИЕ Патологические изменения в эндометрии в пременопаузе сопровождались системным снижением уровня ТФР-β1 и повышением содержания ГдА. В то же время в постменопаузе при патологии эндометрия сопутствовали локальные изменения, представлявшие увеличение экспрессии ФНО-α. Можно предположить, что определение уровней ТФР-β1 и ГдА в сыворотке крови и экспрессии генов ТФР-β1 и ФНО-α в ткани эндометрия может стать полезным дополнением к традиционным методам диагностики патологии эндометрия у больных в пре- и постменопаузе.

Objective: to evaluate changes in T- and B-lymphocyte subpopulations at different stages of rheumatoid arthritis (RA).Patients and methods. The study included 53 patients with a definite RA diagnosis according to the 2010 ACR/EULAR criteria (mean age 54.2 [47; 62] years). Group 1 included 27 patients (25 women and 2 men) without history of synthetic disease modifying anti-rheumatic drugs (sDMARDs) intake, group 2 included 26 patients (22 women and 4 men) receiving sDMARDs (methotrexate or leflunomide). The control group consisted of 29 healthy volunteers (23 women and 6 men), the median age was 58.5 [53; 62] years. In all participants flow cytofluorometry according to the standard technique with immunophenotyping of T- and B-lymphocytes was performed.Results and discussion. Compared to controls, patients in group 1 who had not previously received sDMARDs showed a transient increase in "switched" memory B-cells, transient B-cells, and plasmablasts, which was not observed in patients of group 2 (on sDMARDs therapy). Patients with advanced RA showed a statistically significant decrease in the absolute and relative number of memory B-cells, the absolute and relative number of "switched" B-lymphocytes, as well as the number of plasmablasts and transient cells. In RA patients, a statistically significant rela tionship was established between the number of swollen joints and the level of plasmablasts (r=0.51), memory cells (r=0.54), and "switched" B-cells (r=0.41), p< 0,05 in all cases. There were no statistically significant changes in other subpopulations of B-lymphocytes and the profile of T-lymphocytes.Conclusion. Changes in the B-lymphocyte profile are characteristic of different stages of RA. At an early stage, there is an increase in the number of transient B-lymphocytes, plasmablasts and plasmocytes, and in the advanced stage, a decrease in the level of certain populations of B-lymphocytes, such as memory B-cells and "switched" B-lymphocytes. It can be assumed that the ineffectiveness of sDMARDs is associated with a change in the population composition of B-lymphocytes, which requires further study.

Objective : to evaluate changes in T- and B-lymphocyte subpopulations at different stages of rheumatoid arthritis (RA). Patients and methods . The study included 53 patients with a definite RA diagnosis according to the 2010 ACR/EULAR criteria (mean age 54.2 [47; 62] years). Group 1 included 27 patients (25 women and 2 men) without history of synthetic disease modifying anti-rheumatic drugs (sDMARDs) intake, group 2 included 26 patients (22 women and 4 men) receiving sDMARDs (methotrexate or leflunomide). The control group consisted of 29 healthy volunteers (23 women and 6 men), the median age was 58.5 [53; 62] years. In all participants flow cytofluorometry according to the standard technique with immunophenotyping of T- and B-lymphocytes was performed. Results and discussion . Compared to controls, patients in group 1 who had not previously received sDMARDs showed a transient increase in "switched" memory B-cells, transient B-cells, and plasmablasts, which was not observed in patients of group 2 (on sDMARDs therapy). Patients with advanced RA showed a statistically significant decrease in the absolute and relative number of memory B-cells, the absolute and relative number of "switched" B-lymphocytes, as well as the number of plasmablasts and transient cells. In RA patients, a statistically significant rela tionship was established between the number of swollen joints and the level of plasmablasts (r=0.51), memory cells (r=0.54), and "switched" B-cells (r=0.41), p< 0,05 in all cases. There were no statistically significant changes in other subpopulations of B-lymphocytes and the profile of T-lymphocytes. Conclusion . Changes in the B-lymphocyte profile are characteristic of different stages of RA. At an early stage, there is an increase in the number of transient B-lymphocytes, plasmablasts and plasmocytes, and in the advanced stage, a decrease in the level of certain populations of B-lymphocytes, such as memory B-cells and "switched" B-lymphocytes. It can be assumed that the ineffectiveness of sDMARDs is associated with a change in the population composition of B-lymphocytes, which requires further study.

Introduction. Data about the role of components of adipose tissue in the repair of damaged uterine walls are limited, although a number of authors claim that cell-based drugs from adipose tissue have a positive effect on the repair of damaged uterine walls in laboratory rodents. Meanwhile, stem cells derived from adipose tissue are considered to be the most promising type of cells in regenerative medicine. The aim of the studywas to evaluate the adipocyte components in the uterine wall of rats in healing after a full-thickness surgi-cal incision. Materials and methods. We conducted the study on 40 female Sprague Dawley rats. The animals were sub-jected to a full-thickness longitudinal incision in the wall of the right uterine horn, with the left one serving as an intact control. We carried out morphological examinations of the uterine walls daily in 5 animals from day 1 to 7 and on day 15. The sections from paraffin blocks were stained with hematoxylin and eosin and Mallory’s trichrome staining. Immunohistochemistry detected FABP4+ adipocytes and CD68+ macrophages. The morphometric study was carried out using the Leica system (Leica, Germany). The results underwent processing in the Statistica 12 software (StatSoft). Results. We noticed the period of the most active interaction of adipose tissue with the damaged horn to last from day 3 to 15 and coincide with the macrophage activation in the healing zone. The intact left uterine horn was not involved in the interaction processes with the mesenteric adipose tissue. Starting from day 3 FABP4+ cells in the uterine wall of the operated horn formed groups, creating rounded nest-like structures. Clusters of FABP4+ cells were localized in the healing zone, near the suture material, and in the perime-trium near the mesentery attachment sites. The changes over time of the indicators of the area of cell nests depended on the localization and duration of healing. There were no FABP4+ cells in the left intact horn. Conclusion. We characterized the morphological interaction of adipose tissue with the damaged uterine wall during the first two weeks after a full-thickness surgical incision of the rat uterine horn. The results of the study indicate that adipocytes take an active part in the healing after a surgical incision of the rat uterine wall at the earliest stages. Keywords: rat uterus, hysterotomy, healing, adipocytes, macrophages

We studied the dynamics of morphological changes in the operated segment of the uterine horn of Sprague-Dawley rats during the first 2 weeks of the wound-healing process after a full-thickness surgical incision with regard to the estrous cycle phase. Morphometric parameters of injured uterine right horn were compared with those in the intact left horn of the same animal as a control of changes determined by the hormonal background. It was found that the uterine epithelium in the focus of injury was restored as soon as on day 2 after surgery under the influence of estrous cycle hormones. By day 4, the wound space was completely filled with the endometrial tissue on the side of the uterine lumen and coved by the attached adipose tissue of the mesentery on the side of the abdominal cavity. The thickness of the uterine wall and the uterine lumen differed most strongly between the operated and intact uterine horns during the first 3 days and on day 6 after surgery. The size of the healing area increased during the first three days and reached the peak value by day 3, but then decreased to minimum by day 6.

The clinical efficacy and safety of interleukin-6 (IL-6) receptor blockade have been well studied, but the data on the impact of therapeutic inhibition of IL-6 on B cells are scarce and contradictory. Preliminary reports have shown that B cell function and a humoral immune response may be modulated by an IL-6 receptor inhibitor.Objective: to assess the effect of 12-month tocilizumab (TCZ) therapy on B-cell phenotype and gene expression in RA and to analyze the association between B-cell subsets and RA activity.Subjects and methods. Examinations were made in 24 active RA patients (20 women and 4 men) (median age, 55 [49; 64] years; disease duration, 72 [24; 108] months; DAS28 5.8 [5.3; 6.3]; the patients were seropositive for rheumatoid factor (RF) (100%) and for anti-cyclic citrullinated peptide antibodies (87.3%). The patients received TCZ 8 mg/kg every 4 weeks. After 12 months of therapy, 54% of patients were categorized as good responders, 46% as moderate responders according to the EULAR response criteria. A control group consisted of 29 volunteers (21 women and 8 men; median age, 58.5 [53.0; 62.0] years). Peripheral blood lymphocytes were immunophenotyped at the time of enrollment and after 12 months. The absolute and relative counts of CD19+B lymphocytes, memory B cells (CD19+CD27+), non-switched memory B cells (CD19+IgD+CD27+), switched memory B cells (CD19+IgDCD27+), naive (CD19+IgD+CD27-), double-negative (CD19+IgD-CD27-), transitional (CD19+IgD+CD10+CD38++CD27) B cells, plasma cells (CD19+СD38+), and plasmablasts (CD19+СD38+++IgD-CD27+CD20-) were estimated using multicolor flow cytometry. Results and discussion. The relative and absolute counts of memory B cells (CD19+CD27+) (1.3 [0.9; 1.7]%, 0015 [0.001; 0.003]•109/l), switched memory B cells (CD19+IgD-CD27+) (6.8 [3.6; 11.6]%, 0.01 [0.005; 0.02]•109/l), and the absolute number of transitional B cells (CD19+CD38++CD10+IgD+CD27-) (0.00009 [0; 0.00028]•109/l) were found to be lower in RA patients than in donors: 2.2 [1.1; 3.0]%, 0.003 [0.001; 0.007]•109/l; 12.8 [9.3; 17.0]%, 0.02 [0.01; 0.04]•109/l; 0.0001 [0; 0.0003]•109/l, respectively (p<0.05 for all cases). After 12 months of TCZ therapy initiation, there were decreases in the relative and absolute counts of plasmablasts (CD19+CD38+++CD27+IgD-CD20-) from 0.15 [0.1; 0.3] to 0.1 [0.01; 0.1]% and from 0.0003 [0.00007; 0.004]•109/l to 0.0001 [0; 0.0003]•109/l, respectively (p<0.05). At the same time, the relative and absolute counts of memory B cells (CD19+CD27+) and switched memory B cells (CD19+CD27+IgD-) remained lower in RA patients than in donors: 1.0 [0.7; 1.2] and 2.2 [1.1; 3.0]%; 0.001 [0.006; 0.003]•109/l and 0.003 [0.001; 0.007]•109/l; 3.1 [1.1; 4.2] and 12.8 [9.3; 17.0]%; 0.003 [0.002; 0.006]•109/l and 0.02 [0.01; 0.04]•109/l, respectively (p<0.05 for all cases). Following 12 months of TCZ therapy, the numbers of other B-cell subpopulations were not considerably altered. When included in the study, the patients with RA showed correlations between the absolute count of memory B cells (CD19+CD27+) and the level of C-reactive protein (r=0.50; p<0.05); between the absolute count of plasmablasts (CD19+CD38+++CD27+IgD-CD20-) and the level of RF (r=0.41 and r=0.52; p<0.05). There were no correlations of B cell subsets with clinical and laboratory findings after 12 months of TCZ initiation.Conclusion. Immunophenotyping of peripheral blood B lymphocyte subsets showed the lower relative and absolute counts of memory B cells (CD19+CD27+) and switched memory B cells (CD19+CD27+IgD-) in RA patients than in healthy donors. The found correlations between the counts of memory B cells and plasmablasts and the values of laboratory parameters in patients with high RA activity may suggest that B lymphocytes are involved in the pathogenesis of RA. There was a decline in plasmablast levels after 12 months of TCZ therapy.

The choice of drugs for the treatment of interstitial lung disease (ILD) associated with systemic sclerosis (SS) is currently very limited. Data from a number of studies show that rituximab (RTM) can improve lung function and reduce the severity of skin fibrosis in patients with SS.Objective: to evaluate the efficiency of RTM in a cohort of patients with SS-associated ILD after one-year follow-up. The indications for prescribing RTM were: 1) the inefficiency of standard therapy with glucocorticoids and immunosuppressants (ISs) or the impossibility of their use; 2) the early stage (first 3 years of the disease) with signs of poor prognosis, such as diffuse form, high skin scores (>14), male gender, rapid progression with a significant initial decline in forced vital capacity (FVC) and/or diffusion lung capacity (DLC), and a high anti-Scl-70 antibody positivity.Subjects and methods. The investigators selected a group of patients who had at least two assessment points at a 12-to-18 month interval (the mean follow-up period of 13±2 months) and took at least 1 g of RTM during this period. The investigation included 71 patients with a valid diagnosis of SS. Multi-slice spiral computed tomography (MSCT) revealed ILD in 90% of patients. The disease duration was 5.6±4.4 years. The presence of anti-Scl-70 antibodies was detected in 73% of patients. The mean cumulative dose of RTM was 1.43±0.6 g; 48 patients in Group 1 received ≤2 g of RTM (the mean dose, 1.1±0.1 g) and 23 patients in Group 2 took ≥2 g of RTM (mean dose, 2±0.6 g). Before starting treatment with RTM, all the patients received concomitant therapy with prednisone and 45% - with immunosuppressants.Results and discussion. The results assessed by a physician showed that good and moderate effects of the therapy were observed in 52 (73.2%) and 16 (22.6%) patients, respectively; no effect was seen in 3 (4.2%) patients. Overall, 95.8% of patients reported various degrees of improvement. There were significant changes as reductions in the disease activity index, skin scores, C-reactive protein and IgG levels, the number of patients with a high antinuclear antibody level, and the mean dose of prednisolone as well as increases in an oral aperture size, left ventricular ejection fraction, and 6-minute walk test scores. There were no changes in pulmonary artery systolic pressure and the HAQ DI. FVC increased from 77.35±19.9 to 82.6±20.7% (p=0.001). A minimal clinically significant increase in FVC ≥5% was noted in 41 (57.7%) people. The overall improvement in FVC (ΔFVC) reached 5.24%, while the changes were more significant in Group 2 (ΔFVC 8.98%) than in Group 1 (ΔFVC 3.75%; p=0.01). DLC remained stable, but there were significant group differences: ΔDLC was 3.75% in Group 2 and, conversely, decreased in Group 1 (1.6%; p=0005). The safety profile of the therapy was regarded as good and quite comparable with both the safety profile of ISs and the use of RTM in other trials. Infectious complications were recorded to be most common in 11 (15%) people. Of these, upper respiratory tract infections developed in 7 patients; plantar phlegmon occurred in one case; urinary tract infection and herpes zoster were detected in two and one cases, respectively.The results of this study confirm data from other studies that have demonstrated that RTM exerts a positive effect on SS-associated ILD. We were the first to show the association of positive changes in the measures of pulmonary function tests with the dose of RTM.

Background CD4+ CD25+ Foxp3+ regulatory T (Treg) cells play a key role in maintaining peripheral tolerance and preventing autoimmune disease. Quantitative and/or qualitative deficiencies of Treg have been associated with immune disturbances in systemic lupus erythematosus (SLE). Objectives The main goal of the study was to determine the relationship of CD4+CD25+FoxP3+Treg cells with clinical and immunological manifestations in SLE patients (pts). Methods Frequencies and absolute numbers of peripheral blood CD4+CD25+FoxP3+Treg cells were assessed in 21 healthy donors and 20 SLE pts (2012 SLICC classification criteria); (1M/19F); age 32±13 years; disease duration median (25–75 percentile) 5(1-10) years; SLEDAI2K≥10 - 15(10-22) (14 pts), <10 – 7(6-8) (6 pts). All pts were treated with prednisone, hydroxychloroquine, azathioprine, mycophenolate mofetil, clophosphamide. CD4+CD25+FoxP3+ Treg cells and B-cell subsets were analyzed using multicolor flow cytometry. Results Compared with healthy donors, SLE pts demonstrated significant lower the absolute number of Tregs (0.05; 0.04-0.06 vs 0.03;0.02-0.05x109/L, p<0,036), with a high percentage of Treg (8.8;7.5-10.5 vs 12.0;8.8-17.0%, p<0.02). The median percentage of Tregs was lower in pts with acute SLE compared to chronic SLE pts (9.0;8.3-9.9 vs 13.5;12.7-18.7%, p<0,02). SLE pts with high activity had a lower frequencies of Tregs (10.2;8.5-15.0 vs 15.8;12.7-20.4%, r=-0,51, p<0,05). Low count of Tregs correlated with elevated level of IgG (r=-0,52, p<0,05). Absolute number of Tregscorrelated negatively with percentage and absolute count of transitional (CD19+IgD+CD10+CD38++CD27-) B cells (r=-0,66 and r=-0,63, p<0,05). Conclusion Decreased amount of T regs in SLE is associated with high disease activity, acute course and expansion of autoreactive B cells. References [1] Zhang SX, Ma XW, Li YF, Lai NL, Huang ZH, Fan K, Wang CH, Li XF. The Proportion of Regulatory T Cells in Patients with Systemic Lupus Erythematosus: A Meta-Analysis. J Immunol Res. 2018 Sep 3;2018:7103219. doi: 10.1155/2018/7103219. [2] Żabińska M, Krajewska M, Kościelska-Kasprzak K, Jakuszko K, Bartoszek D, Myszka M, Klinger M. CD4(+)CD25(+)CD127(-) and CD4(+)CD25(+)Foxp3(+) Regulatory T Cell Subsets in Mediating Autoimmune Reactivity in Systemic Lupus Erythematosus Patients. Arch Immunol Ther Exp (Warsz). 2016 Oct;64(5):399-407. doi: 10.1007/s00005-016-0399-5. Disclosure of Interests None declared