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The intensity of CD56 receptor expression by NK-92 cells (a,b) and peripheral blood NK cells (c,d). Cells were cultured without inducers (grey) and with TGFβ (dark green). Isotype control is shown in black.
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During pregnancy, uterine NK cells interact with trophoblast cells. In addition to contact interactions, uterine NK cells are influenced by cytokines, which are secreted by the cells of the decidua microenvironment. Cytokines can affect the phenotypic characteristics of NK cells and change their functional activity. An imbalance of pro- and anti-in...
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... It was also found that a short-term (24 hours) preincubation of NK cells with TGFβ does not affect the cytotoxicity of NK cells toward trophoblasts. However, preincubation with TGFβ for 96 hours resulted in stimulation of NK cell cytotoxicity toward trophoblasts [30]. To date, there are no published data on the participation of CD105, or its soluble form, in the regulation of the abovementioned receptors or the cytotoxicity of NK cells toward trophoblasts. ...
... To test the method, the cells were preincubated with the induc tors for 30 minutes, 4, and 6 hours. Based on the Yes [42] Yes [43,45] No data No data Natural killer cell marker NKG2A Yes [21] Yes [30,46] No data No data Inhibitory receptor [47], CD94 co-receptor, binds HLA-E ...
... Yes [21] Yes [30,46] No data No data Inhibitory receptor, NKG2A co-receptor, together bind HLA-E [47] NKG2D Yes [48] Yes [49] No data No data Activatory receptor [47], binds MICA/B ...
The interaction of natural killer and trophoblast cells underlies maternal-fetal immune tolerance. The data on the participation of endoglin (ENG, CD105), or its soluble form, in the regulation of the communication of these cells are currently insufficient. In this study, we have investigated the role of endoglin in the intercellular interactions between natural killer cells and trophoblasts. Here, we show that NK-92 cells and JEG-3 cells constitutively express MICA/B, and CD105. In the presence of JEG-3 cells, the expression of NKG2D, CD94, MICA/B, and CD105 by NK-92 cells was increased, and the number of NK-92 cells expressing NKG2A, CD94, and MICA was reduced. Antibodies against ENG and recombinant endoglin (rENG), attenuated the trophoblasts’ influence and returned the phenotype of the NK-92 cells to that of cells found in monoculture conditions. The antibodies and rENG also increased the expression of pSMAD2/3 by NK cells in both monoculture and co-culture conditions. The antibodies increased the trophoblasts’ sensitivity to the cytotoxic effect of NK cells. In general, our findings indicate a significant role of endoglin in the intercellular communication between NK cells and trophoblasts. We also speculate that endoglin forms a complex with TGFβ, which aids in TGFβ trafficking between these cells.
... The first trimester is characterized by a highest frequency of spontaneous abortions due to onset of HLA molecule expression by trophoblasts and luteo-placental shift [13]. At the same time, the cytotoxic potential of NK cells and the production of proinflammatory cytokines decrease, while the produc tion of anti-inflammatory cytokines increases [14][15][16][17]. Regulatory peripheral blood NK cells migrate in early pregnancy to the uterus, where they form a pool of decidual NK cells, which are the main effectors of immune tolerance in the fetoplacental junctional zone, limit cytotoxic responses to fetal antigens, and produce factors that promote trophoblast invasion [15,18,19]. ...
... At the same time, the cytotoxic potential of NK cells and the production of proinflammatory cytokines decrease, while the produc tion of anti-inflammatory cytokines increases [14][15][16][17]. Regulatory peripheral blood NK cells migrate in early pregnancy to the uterus, where they form a pool of decidual NK cells, which are the main effectors of immune tolerance in the fetoplacental junctional zone, limit cytotoxic responses to fetal antigens, and produce factors that promote trophoblast invasion [15,18,19]. An increase in the total count and cyto toxic activity of peripheral blood and decidual NK cells is associated with spontaneous abortions and other complications of pregnancy (preeclampsia) [14,19,20]. ...
... An increase in the total count and cyto toxic activity of peripheral blood and decidual NK cells is associated with spontaneous abortions and other complications of pregnancy (preeclampsia) [14,19,20]. In the third trimester of pregnancy, maternal immunoreactivity is determined by preparing for labor and recoverying altered functions [13,15,19]. ...
... В первом триместре клетки трофобласта начинают экспрессировать молекулы НLA и происходит лютеоплацентарная смена, что является причиной высокой частоты спонтанных абортов [9]. Цитотоксический потенциал и продукция провоспалительных цитокинов NK-клетками периферической крови снижаются [2,5,7]. Регуляторные NK-клетки в ранние сроки беременности мигрируют из периферической крови в матку, где трансформируются в децидуальные NK-клетки и подавляют цитотоксические реакции к антигенам плода, продуцируют факторы роста трофобласта [2,5,7]. ...
... Цитотоксический потенциал и продукция провоспалительных цитокинов NK-клетками периферической крови снижаются [2,5,7]. Регуляторные NK-клетки в ранние сроки беременности мигрируют из периферической крови в матку, где трансформируются в децидуальные NK-клетки и подавляют цитотоксические реакции к антигенам плода, продуцируют факторы роста трофобласта [2,5,7]. В III триместре изменения иммунореактивности связаны с подготовкой к родам [2,5,7]. ...
... Регуляторные NK-клетки в ранние сроки беременности мигрируют из периферической крови в матку, где трансформируются в децидуальные NK-клетки и подавляют цитотоксические реакции к антигенам плода, продуцируют факторы роста трофобласта [2,5,7]. В III триместре изменения иммунореактивности связаны с подготовкой к родам [2,5,7]. Нарастание количества и цитотоксичности NK-клеток при беременности является одной из причин спонтанных абортов и других патологий беременности (преэклампсии) [1,8,9]. ...
РЕЗЮМЕ
Цель-оценить метаболическую активность разных субпопуляций натуральных киллеров (NK-клеток) периферической крови в I и в III триместрах физиологически протекающей беремен-ности (ФПБ). Материал и методы. В исследование включены здоровые женщины в I или III триместрах ФПБ. Группу сравнения составили условно-здоровые небеременные женщины в фолликулярной фазе менструального цикла.
Объектом исследования являлась периферическая кровь. Метаболическую активность оценивали по экспрессии белка-транспортера глюкозы Glut-1 и включению митохондриального зонда Mito-Spy Green, отражающего массу и объем митохондриального компартмента, методом проточной цитофлюориметрии на регуляторных (CD56 bright CD16-), цитотоксических (CD56 dim CD16 +), минорных цитотоксических (CD56-CD16 hi) NK-клетках.
Результаты и обсуждение. У небеременных количество метаболически активных клеток, экспрес-сирующих Glut-1 и имеющих большую массу и объем митохондрий (Mito +), значительно выше в субпопуляции цитотоксических CD56-CD16 hi NK-клеток, чем среди регуляторных CD56 bright CD16-и цитотоксических CD56 dim CD16 + NK-клеток. В I тримеcтре беременности уровень Glut-1 + Mito + ре-гуляторных CD56 bright CD16-и цитотоксических CD56 dim CD16 + NK-клеток значительно нарастает по сравнению с таковым у небеременных, тогда как процент цитотоксических Glut-1 + Mito + CD56-CD16 hi NK-клеток остается высоким, но не меняется. В III триместре количество Glut-1 + Mito + регу-ляторных CD56 bright CD16-NK-клеток остается выше, чем у небеременных. Количество цитотокси-ческих Glut-1 + Mito + CD56 dim CD16 + NK-клеток снижается по сравнению с таковым у небеременных, а Glut-1 + Mito + CD56-CD16 hi NK-по отношению к I триместру.
Заключение. При ФПБ увеличивается количество метаболически активных цитотоксических и ре-гуляторных NK-клеток периферической крови. Ключевые слова: натуральные киллеры, беременность, Glut-1, масса митохондрий.
... Наиболее активно изучают их взаимодействие с клетками трофобласта. Показано, что клетки взаимно регулируют характеристики друг друга как за счет контактных, так и дистантных взаимодействий, обеспечивая формирование оптимального микроокружения для развивающегося плода [104,162]. На сегодняшний день считают, что взаимодействие между NK-клетками и клетками трофобласта -ключевое звено в наступлении и развитии беременности, нарушение которого приводит к репродуктивным патологиям. Причем взаимодействие может быть нарушено и изменением числа NK-клеток [4], и усилением [46] либо ингибированием их цитотоксической активности [170], и изменением спектра цитокинов [41]. ...
Natural killer (NK) cells represent one of the innate lymphoid cell subsets, which are often studied in the context of antitumor and antiviral immunity, as well as due to their localization in the zone of the mother-fetus contact (in the uterus), therefore underlying their extensive investigation in developing pregnancy. At the same time, their role in antibacterial immune response has been poorly examined. Because NK cells can produce cytokines, one of putative options for their participation in eliminating prokaryotic pathogens may be coupled to regulation of immune system cells such as dendritic cells, macrophages, etc. However, there have been also described variants of contact cytolysis of cells infected with intracellular bacteria enabled due to cytotoxic proteins perforin, granzymes, granulisin found in NK cells. In recent years, it has become known that NK cells take part in development of immune response against extracellular bacteria including the ESKAPE group bacteria, which includes opportunistic prokaryotes that most actively develop antibiotic resistance and cause nosocomial infections. Here, we attempted to review the data on the role NK cells play in antibacterial immunity. Assessing a crosstalk between ESKAPE group bacteria and NK cells also attracts researchers due to the ability of prokaryotes to alter functions of immune cells, but very little is known about the effects they exert on NK cells. At the same time, such data could be applied to seek out for new ways to treat oncological diseases as well as pave the basis for new approaches to regulating NK cell characteristics in reproductive pathologies. As mentioned earlier, the latter occur in the decidual membrane, where they can interact with fetal cells including trophoblast cells. It is believed that cells can mutually regulate each others properties necessary for the course of physiological pregnancy. Probably, imbalance in this system can lead to development of reproductive pathologies. The review summarizes the currently available data on the effects of ESKAPE group bacteria on NK cells, and also considers putative mechanisms for emergence of impaired interaction between NK cells and trophoblasts exposed to ESKAPE group bacteria. Owing to few publications available on this phenomenon, the experimental study assessing an impact of ESKAPE group bacteria on NK cell properties is envisioned as a necessary stage in development of contemporary biology.
... There are a considerable number of dNK cells in the decidua, accounting for approximately 70% of the total number of deciduous lymphocytes [27]. During pregnancy, the decidua maintains close contact with trophoblasts without causing damage because trophoblasts exert an immunosuppressive effect on dNK cells [28]. NK cells in the decidua are a dominant subpopulation, and various factors with inhibitory effects on NK cells are locally present. ...
Decidual natural killer cells (dNK cells) are an essential component of the immune cells present at the maternal–foetal interface during early pregnancy, and they play a vital role in various physiological processes. Abnormalities in the ratio or function of dNK cells have been linked to recurrent miscarriages. CD96 has been previously shown to regulate NK cell function in the tumour microenvironment; however, its role and mechanism at the maternal–foetal interface remains unclear. The present study aimed to investigate the immunomodulatory role of CD96 in dNK cells and its function at the maternal–foetal interface. Immunofluorescence staining and flow cytometry were used to detect the expression of cellular markers such as CD96. Furthermore, the secretory function, adhesion-function-related molecules, and cell proliferation markers of CD96+ and CD96− dNK cells were detected using flow cytometry. In addition, we performed cell culture experiments via the magnetic bead sorting of NK cells to detect changes in the expression of the aforementioned functional molecules in dNK cells after the CD96 blockade. Furthermore, we examined the functional characteristics of dNK cells after palmitic acid treatment at a concentration of 10 μM. We also examined the changes in dNK cell function when subjected to the combined effect of palmitic acid and CD96 antagonists. The results indicated that CD96, TIGIT, CD155, and CD112 were highly expressed at the maternal–foetal interface, with dNK cells predominantly expressing CD96, whereas TIGIT was mainly expressed on T cells, and CD155 and CD112 were mainly present in metaphase stromal and trophoblast cells. CD96+ dNK cells displayed low cytotoxic activity and a high adhesion phenotype, which mediated the immunosuppressive effect on dNK cells at the maternal–foetal interface. Palmitic acid upregulated CD96 expression on the surface of dNK cells in the coculture system, inhibiting dNK cell activity and increasing their adhesion molecule expression. CD96 antagonist treatment blocked the inhibitory effect of trophoblasts on dNK cells, resulting in enhanced cytokine secretion and reduced adhesion. The results of this study provide valuable insight into the immunomodulatory role of CD96 in dNK cells and its mechanism at the maternal–foetal interface, particularly in metaphase NK cells. This study sheds light on the mechanisms of immune regulation at the maternal–foetal interface and their implications for the study of recurrent miscarriages of unknown origin.
... Cytokines are critical determinants of the phenotypic characteristics and functional activity of NK cells in immune response and development. In the article entitled "Proand Anti-Inflammatory Cytokines in the Context of NK Cell-Trophoblast Interactions", Mikhailova et al. [5] introduced the effects of cytokines on NK cells in the presence of trophoblasts. Cytokines in the uteroplacental complex play important roles in the phenotype and functional state of NK cells during contact interaction with trophoblasts. ...
Natural killer (NK) cells are innate immune cells that demonstrate cytolytic activity against tumor cells, virus-infected cells and other physiologically stressed cells, such as senescent cells [...]
... Both trophoblasts and natural killers have a wide arsenal for mutual containment, since trophoblasts are considered foreign elements by the mother's immune system. On the other hand, the invasion of trophoblasts into the endometrium is accompanied by their influence on cells of the microenvironment, including endometrial, endothelial, and maternal NK cells [94][95][96][97]. Despite this, NK cells actively control the proliferation and migration of trophoblasts under conditions of physiological pregnancy, which in turn restrains the excessive cytotoxicity of NK cells [98][99][100][101]. ...
The interaction of natural killer (NK) and trophoblast cells underlies the formation of immune tolerance in the mother–fetus system and the maintenance of the physiological course of pregnancy. In addition, NK cells affect the function of trophoblast cells, interacting with them via the receptor apparatus and through the production of cytokines. Microvesicles (MVs) derived from NK cells are able to change the function of target cells. However, in the overall pattern of interactions between NK cells and trophoblasts, the possibility that both can transmit signals to each other via MVs has not been taken into account. Therefore, the aim of this study was to assess the effect of NK cell-derived MVs on the phenotype, proliferation, and migration of trophoblast cells and their expression of intracellular messengers. We carried out assays for the detection of content transferred from MV to trophoblasts. We found that NK cell-derived MVs did not affect the expression of CD54, CD105, CD126, CD130, CD181, CD119, and CD120a receptors in trophoblast cells or lead to the appearance of CD45 and CD56 receptors in the trophoblast membrane. Further, the MVs reduced the proliferation but increased the migration of trophoblasts with no changes to their viability. Incubation of trophoblast cells in the presence of MVs resulted in the activation of STAT3 via pSTAT3(Ser727) but not via pSTAT3(Tyr705). The treatment of trophoblasts with MVs did not result in the phosphorylation of STAT1 and ERK1/2. The obtained data indicate that NK cell-derived MVs influence the function of trophoblast cells, which is accompanied by the activation of STAT3 signaling.
... CD56 dim CD16 + NK cell is a main subset in peripheral blood and mediates natural and antibody-dependent cellular cytotoxicity, while CD56 bright CD16 -NK cell subset has a higher secretory ability and lower cytotoxicity (Uppendahl et al., 2017, Jin et al., 2021. Although a few researches showed that the proportion of CD56 bright CD16 -NK cells didn't change after cocultured with the human choriocarcinoma cell Abbreviations: ADCC, Antibody-dependent cell-mediated cytotoxicity; CC, Choriocarcinoma; CM, conditional medium; dl-THP, dl-tetrahydropalmatine; DMSO, dimethyl sulfoxide; GTN, gestational trophoblastic neoplasia; hCG, human chorionic gonadotropin; LDH, lactate dehydrogenase; PBMCs, peripheral blood mononuclear cells; PI, propidium iodide; pNK, peripheral NK cells; THB, tetrahydroberberine; THC, tetrahydrocoptisine. lines JEG-3 (Mikhailova et al., 2020), along with decreased proportion of CD56 + NK cells (Mikhailova et al., 2022). How about the proportion of CD56 dim CD16 + NK cells and CD56 bright CD16 -NK cells in CM (medium from JEG-3) are still unknown. ...
... Several studies have indicated that tumor microenvironment can change the phenotype and damage NK cell cytotoxic functions by downregulating activating receptors or upregulating inhibitory receptors (Bruno et al., 2014, Bruno et al., 2013, Han et al., 2018. Culturing pNK cells with JEG-3 in the presence of TGF-β resulted in a significant decrease in the percentage of NKp44 + cells in pNK, while the percentage of NKp44 + cells hardly changed when the two were co-cultured in the absence of TGF-β (Mikhailova et al., 2022). Meanwhile, co-culture of JEG-3 and NK92 allows expression intensity of the receptors on the surface of NK92 cells that resemble decidual NK phenotypes, such as NKG2A, NKp30, NKp44, KIR2DL1, KIR2DL3, KIR2DL4, CD117, CD215 and CD127 (Mikhailova et al., 2021). ...
Natural killer cell-based immunotherapy has become a leading-edge tool against cancer, but still faces a variety of challenges, such as phenotype shift and dysfunction of NK cells in tumor microenvironment. Thus, finding potent agents that could inhibit the phenotype shift and incapacity of NK cells in the tumor microenvironment is essential for improving antitumor effects. dl-tetrahydropalmatine (dl-THP), one of the active alkaloids of Chinese herb Corydalis Rhizoma, has been proven to possess antitumor activity. However, whether dl-THP acts on NK cells to enhance antitumor activity remains unknown. In this study, we found that the proportion of blood CD56dimCD16+ NK cells was decreased while the proportion of CD56brightCD16- NK cells was increased when the cells were cultured in conditional medium (CM, medium from the human choriocarcinoma cell lines JEG-3). dl-THP could alter the varied proportion of CD56dimCD16+ NK cells and CD56brightCD16- NK cells in CM respectively. Importantly, the expression level of NKp44 on CD56dimCD16+ NK cells was dramatically reduced when the cells were cultured in CM, which could also be reversed by dl-THP. Furthermore, dl-THP increased the decreased NK-cell cytotoxicity when cells were cultured in CM. In summary, our study demonstrated that dl-THP could recover the decreased NKp44 expression level on CD56dimCD16+ NK cells and restore the cytotoxicity of NK cells in tumor microenvironment.
The functions of peripheral blood NK cells change significantly during pregnancy, which is mainly due to the inhibition of their cytotoxicity. The functional activity of NK cells is directly related to their metabolic status, but these changes in physiological pregnancy have not been studied. The aim of this work is to study the expression of Glut-1, CD94 and CD107a molecules characterizing metabolic and cytotoxic activity, as well as the mitochondrial mass of different subpopulations of peripheral blood NK cells in the I and III trimesters of physiological pregnancy. The object of the study was the peripheral blood of healthy women in the I and III trimesters of physiological pregnancy. The control group consisted of healthy non-pregnant women in the follicular phase of the menstrual cycle. The expression of Glut-1, CD94, CD107a molecules and the mitochondrial mass were analyzed by flow cytometry on regulatory (CD16–CD56bright), cytotoxic (CD16+CD56dim), minor cytotoxic (CD16hiCD56–) NK cells. It was found that in non-pregnant women, minor cytotoxic CD16hiCD56–NK have the highest expression of Glut-1, CD107a and the lowest expression of CD94 compared to other NK cell subpopulations. On regulatory CD16–CD 56bright and cytotoxic CD16+CD56dimNK, the expression of these molecules is comparable to each other. The mitochondrial mass is similar in all studied subpopulations. In the first trimester, the expression of Glut-1 increases on regulatory CD16–CD56brightNK, the mitochondrial mass and the expression of CD94, CD107a in all NK cells do not differ from non-pregnant ones. In the third trimester, the mitochondrial mass increases in cytotoxic CD16+CD56dimNK cells, but CD94 expression decreases compared to non-pregnant ones, and the expression CD94 in regulatory CD16–CD56brightNK increases compared to the first trimester. CD107a expression in minor cytotoxic CD16hiCD56–NK decreases, but in other subpopulations does not change compared to non-pregnant. The expression of Glut-1 does not change in all subpopulations. Thus, different subpopulations of peripheral blood NK cells are heterogeneous in the expression of Glut-1, CD107a, CD94. The expression of these molecules during physiological pregnancy varies by trimester. The obtained results are important for understanding the mechanisms of NK cell function regulations during pregnancy.
