Expression of arginase 1 and tyrosine kinase Mer by blood monocytes in the dynamics of physiological pregnancy

During pregnancy, the maternal immune system must maintain tolerance to paternal antigens, at the same time being able to eliminate pathogens, which is achieved by the weakening of adoptive immunity and the activation of innate immunity, in particular, monocytes. However, the question about the functional phenotype of monocytes, having not only pro-inflammatory, but also anti-inflammatory activity, remains open. In the given work, we have investigated the expression of M2-associated suppressive markers Arg1 and MerTK in monocyte subpopulations during uncomplicated pregnancy. Fifty-three pregnant women with uncomplicated gestation were recruited, including 14 pregnant in the 1st trimester, 20 – in the 2nd and 19 – in the third pregnancy trimester. The comparison group consisted of 15 fertile unpregnant women without aggravated somatic anamnesis, with a history of at least one childbirth. The findings showed that in the unpregnant group circulating Mo express Arg1 and MerTK, and the most relative number of Arg1+ and MerTK+ cells is concentrated in intermediate and nonclassic monocytes. During pregnancy the expression of researched molecules in monocytes reliably increases. An increase in MerTK expression is manifested by a simultaneous increase in the number of MerTK+ cells and the mean fluorescence intensity of this marker; it is observed in the 1st and 2nd trimesters and registered in all three monocyte subpopulations. At the same time, an increase in Arg1 expression is manifested either by an enhancement of Arg1+ cells, or an increase in receptor density; it is registered throughout pregnancy, including the 3rd trimester, and is maximally expressed in classic monocytes. There is a direct correlation between the number of Arg1+ and MerTK+ cells in intermediate Mo, which increases with the progression of pregnancy, and in the 3rd trimester is also detected in classical and non-classical Mo. In general, the revealed increase in the expression of Arg1 and MerTK by monocytes indicates an increase in the anti-inflammatory potential of monocytes during pregnancy, and the involvement of monocytes in the regulation of the inflammatory process at the system level. Moreover, the features of Arg1 and MerTK expression in various monocyte subpopulations during pregnancy suggest that monocytes expressing Arg1 and MerTK can mediate different mechanisms of immune adaptation during pregnancy

Expression of M2-associated molecules in circulating monocyte subsets in fertile non-pregnant women and pregnant women with uncomplicated pregnancy

In humans circulating monocytes include classical (CD14++CD16- ), intermediate (CD14++CD16+) and non-classical/alternative (CD14+CD16++) monocytes, which in turn can be activated via the classical or alternative pathway. Pregnancy is accompanied by significant changes in the monocyte compartment, which is manifested by an increase in the number of circulating monocytes, including the proportion of intermediate monocytes, and a change in their function. However, the functional properties of monocyte subsets during gestation remain largely unexplored. We hypothesized that circulating monocytes may be activated in an alternative pattern and acquire features of M2 polarization (anti-inflammatory / immunosuppressive properties). The aim of the investigation was to study M2-associated markers that characterize the anti-inflammatory and immunosuppressive potential of myeloid cells in subpopulations of circulating monocytes in fertile nonpregnant women and women with uncomplicated pregnancy in the 2nd trimester. It was shown that in fertile non-pregnant women intermediate and non-classical monocytes are characterized by a higher expression of M2-associated markers (CD206, Arginase 1, MerTK) compared to classical monocytes. In the 2nd trimester of pregnancy, the expression of these molecules on monocytes increases significantly, which is manifested by 1) an increase in the proportion of CD206+ cells in subpopulations of classical and intermediate monocytes, 2) an increase in the mean fluorescence intensity of Arginase 1 in all monocyte subsets, 3) an increase in the proportion of MerTK+ cells in subpopulations of classical and intermediate monocytes and mean fluorescence intensity across all monocyte subsets. The highest content of CD206+ and MerTK+ cells in pregnant women is detected in the subpopulation of intermediate monocytes, and the highest values of the mean fluorescence intensity of Arginase 1 and MerTK – in the subpopulations of intermediate and non-classical monocytes. The data obtained demonstrate that monocytes of pregnant women in the 2nd trimester of pregnancy are characterized by signs of M2 polarization. This is confirmed not only by an increase in the expression of the M2-associated mannose receptor CD206, but also by an increase in the expression of Arginase 1 and MerTK, which mediate the immunosuppressive activity of myeloid cells and, in particular, macrophages of the M2 phenotype. Further studies of M2-associated markers in monocyte subpopulations during gestation will allow a more detailed characterization of the regulatory role of circulating myeloid cells during pregnancy

Фактор роста плаценты модулирует ответ активированных in vitro T-клеток

Background. Recent studies demonstrated immunosuppressive properties of vascular endothelial growth factor (VEGF-A) and identified VEGF-A as a key mediator of tumor-induced immunosuppression. Placental growth factor (PlGF) is another member of VEGF family in which dramatic elevation is associated with effective immune adaptation in successful pregnancy, whereas low concentrations are related to pregnancy complications resulting from the activation of immune system. Previously, we have shown that activated T cells express VEGF receptor type 1 (VEGFR-1), and PlGF inhibits T cell proliferation in VEGFR-1–dependent manner.The aim of the present study was to further characterize PlGF effects on T cell responses in vitro.Materials and methods. Peripheral blood mononuclear cells (PBMC) from healthy donors were stimulated with anti-CD3 monoclonal antibodies (a-CD3) in the absence or presence of PlGF and assessed for IL-10 production, programmed cell death and the expression of inhibitory receptors (PD-1, CTLA-4, Tim-3) in CD4+ and CD8+ T cell subsets.Results. The addition of PlGF to PBMC cultures activated with a-CD3 resulted in increased percentages of IL- 10-producing CD4+ and CD8+ T cells. Besides, PlGF promoted CD8+ T cells apoptosis while did not affect programmed cell death within CD4+ T cells. Notable, T cell activation with a-CD3 in the presence of PlGF was accompanied by the enhancement of PD-1-expressing cells in CD8+ T cell subset and Tim-3-expressing cells in both CD4+ and CD8+ T cells, and by the increased expression of PD-1 and Tim-3 on T cells.Conclusion. Our in vitro findings indicate that PlGF can inhibit T cell responses due to the increasing interleukin-10 (IL-10) production, promoting CD8+ T cell apoptosis and enhancing the expression of PD-1 and Tim-3 inhibitory receptors. Given the elevated levels of PlGF in successful pregnancy and its decrease in gestation complications, the obtained data also suggest that PlGF-mediated suppression may be implicated into the governing immune evasion in pregnancy.Актуальность. Недавние исследования выявили иммуносупрессивные свойства фактора роста эндотелия сосудов (VEGF-A) и его ключевую роль в опухоль-индуцированной иммуносупрессии. Плацентарный фактор роста (PlGF) является еще одним представителем семейства VEGF, резкое возрастание которого ассоциировано с эффективной иммунной адаптацией при успешной беременности, тогда как низкие  концентрации PlGF являются предиктором гестационных осложнений на фоне активации  иммунной системы. Ранее нами показано, что активированные Т-клетки экспрессируют рецепторы VEGF 1-го типа (VEGFR-1) и PlGF через связывание с VEGFR-1 ингибирует пролиферацию Т-клеток.Цель. Дальнейшее изучение влияния PlGF на T-клеточный ответ in vitro.Материалы и методы. Мононуклеарные клетки (МНК) периферической крови здоровых доноров стимулировали моноклональными анти-CD3-антителами (a-CD3) в отсутствие и присутствии рекомбинантного PlGF и оценивали продукцию интерлейкина-10 (IL-10), уровень апоптоза и экспрессию ингибиторных рецепторов (PD-1, CTLA-4, Tim-3) в  субпопуляциях CD4+ и CD8+ T-клеток. Результаты. Активация МНК a-CD3 в присутствии PlGF приводила к возрастанию относительного содержания CD4+ и CD8+ T-клеток, продуцирующих IL-10. Кроме того, PlGF усиливал апоптоз активированных CD8+ T-лимфоцитов, не влияя значимо на уровень программированной клеточной гибели CD4+ Т-клеток. Характерно, что активация Т-клеток a-CD3 в присутствии PlGF сопровождалась возрастанием PD-1  экспрессирующих клеток в субпопуляции CD8+ Т-клеток и Tim-3-экспрессирующих клеток среди CD4+ и CD8+ Т-клеток, а также повышением уровня экспрессии PD-1 и Tim-3 на Т-клетках.Заключение. PlGF способен ингибировать Т-клеточный ответ посредством усиления продукции IL-10 и активационно-индуцированного апоптоза CD8+ Т-клеток, а также экспрессии ингибиторных рецепторов. Учитывая повышенный уровень PlGF при физиологической беременности и его снижение при гестационных осложнениях,  полученные данные позволяют предполагать, что ингибиторный эффект PlGF на Т-клеточный ответ может являться еще одним механизмом, обеспечивающим защиту плода  от иммунной системы матери

Who does not gain weight? Prevalence and predictors of weight maintenance in young women

OBJECTIVE: To investigate the prevalence and predictors of weight maintenance over time in a large sample of young Australian women. DESIGN: This population study examined baseline and 4 y follow-up data from the cohort of young women participating in the Australian Longitudinal Study on Women\u27s Health. SUBJECTS: A total of 8726 young women aged 18-23 y at baseline. MEASURES: Height, weight and body mass index (BMI); physical activity; time spent sitting; selected eating behaviours (eg dieting, disordered eating, takeaway food consumption); cigarette smoking, alcohol consumption; parity; and sociodemographic characteristics. RESULTS: Only 44% of the women reported their BMI at follow-up to be within 5% of their baseline BMI (maintainers); 41% had gained weight and 15% had lost weight. Weight maintainers were more likely to be in managerial or professional occupations; to have never married; to be currently studying; and not to be mothers. Controlling for sociodemographic factors, weight maintainers were more likely to be in a healthy weight range at baseline, and to report that they spent less time sitting, and consumed less takeaway food, than women who gained weight. CONCLUSIONS: Fewer than half the young women in this community sample maintained their weight over this 4 y period in their early twenties. Findings of widespread weight gain, particularly among those already overweight, suggest that early adulthood, which is a time of significant life changes for many women, may be an important time for implementing strategies to promote maintenance of healthy weight. Strategies which encourage decreased sitting time and less takeaway food consumption may be effective for encouraging weight maintenance at this life stage.<br /

Свободная ДНК в фолликулярной жидкости у женщин с различными показателями овариальной функции

The aim of the study was to evaluate cell-free DNA (cfDNA) in the follicular fluid (FF) of women undergoing IVF treatment and to analyze the relationship between cfDNA levels and the parameters of folliculogenesis and oogenesis as well as the quality of embryos. Materials and methods. The study included 53 women aged 20 to 45 years. In 49 patients, oocytes were obtained by stimulating ovulation with gonadotropins, and 4 patients underwent natural cycle IVF without hormonal stimulation. Measurement of cfDNA was carried out by fluorimetry using QuantiFluor™ Handheld Fluorometers (BioSilica, Russian Federation). Results. The FF of women with ovulation stimulation revealed a higher level of cfDNA as opposed to FF of women in the natural cycle. There were no differences in the cfDNA levels in women with infertility and oocyte donors. Women with infertility lasting for more than 5 years had a higher level of cfDNA. Women with the elevated anti-Mullerian hormone (AMH) levels were characterized by the high FF cfDNA concentration and a large number of follicles. Likewise, correlation analysis showed that FF cfDNA was significantly and positively correlated with the AMH level. The obtained data revealed the participation of cfDNA in different stages of oogenesis. Conclusions. The level of FF cfDNA in women may serve as an additional biomarker of the effectiveness of ovulation induction.Целью работы явились исследование свободной ДНК (свДНК) в фолликулярной жидкости (ФЖ) женщин, проходящих лечение по программе экстракорпорального оплодотворения (ЭКО), и анализ взаимосвязи уровня свДНК с различными клиническими параметрами, а также показателями овариального резерва, фолликуло- и оогенеза.Материалы и методы. В исследование были отобраны 53 женщины, у 49 из которых ооциты получали при стимуляции овуляции гонадотропинами, у 4 – в естественном цикле. Концентрация свободной ДНК оценивалась флуориметрическим методом с использованием прибора QuantiFluor™ Handheld Fluorometers (BioSilica, Россия).Результаты. Образцы ФЖ содержали детектируемые концентрации свДНК, уровень которой у женщин со стимулированной овуляцией был значимо выше, чем у женщин в естественном цикле. Различий в содержании свДНК у женщин с бесплодием и доноров ооцитов не отмечалось. В то же время у женщин с более длительным периодом бесплодия (&gt;5 лет) концентрация свДНК была выше, чем в оппозитной группе. Женщины с повышенным уровнем антимюллерового гормона, характеризовались более высоким содержанием свДНК, при этом между уровнем антимюллерового гормона и концентрацией свДНК выявлялась умеренная, но достоверная корреляционная зависимость. Наиболее высокое содержание свДНК выявлялось также в группе женщин с более высоким количеством овуляторных фолликулов. В то же время концентрация свДНК не коррелировала с количеством ооцитов.Заключение. Уровень свДНК в ФЖ женщин может рассматриваться как дополнительный критерий эффективности ответа яичников на стимуляцию овуляции

Effects of a Caffeine-Containing Energy Drink on Simulated Soccer Performance

[Background] To investigate the effects of a caffeine-containing energy drink on soccer performance during a simulated game. A second purpose was to assess the post-exercise urine caffeine concentration derived from the energy drink intake. [Methodology/Principal Findings] Nineteen semiprofessional soccer players ingested 630±52 mL of a commercially available energy drink (sugar-free Red Bull®) to provide 3 mg of caffeine per kg of body mass, or a decaffeinated control drink (0 mg/kg). After sixty minutes they performed a 15-s maximal jump test, a repeated sprint test (7×30 m; 30 s of active recovery) and played a simulated soccer game. Individual running distance and speed during the game were measured using global positioning satellite (GPS) devices. In comparison to the control drink, the ingestion of the energy drink increased mean jump height in the jump test (34.7±4.7 v 35.8±5.5 cm; P<0.05), mean running speed during the sprint test (25.6±2.1 v 26.3±1.8 km · h−1; P<0.05) and total distance covered at a speed higher than 13 km · h−1 during the game (1205±289 v 1436±326 m; P<0.05). In addition, the energy drink increased the number of sprints during the whole game (30±10 v 24±8; P<0.05). Post-exercise urine caffeine concentration was higher after the energy drink than after the control drink (4.1±1.0 v 0.1±0.1 µg · mL−1; P<0.05). [Conclusions/significance] A caffeine-containing energy drink in a dose equivalent to 3 mg/kg increased the ability to repeatedly sprint and the distance covered at high intensity during a simulated soccer game. In addition, the caffeinated energy drink increased jump height which may represent a meaningful improvement for headers or when players are competing for a ball