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  • 1
    In: Medical Immunology (Russia), SPb RAACI, Vol. 21, No. 4 ( 2019-10-29), p. 669-688
    Abstract: Natural killer (NK) cells are of special interest among a multitude of microvesicle (MV) source cells. NK cells are a lymphocyte subpopulation performing contact cytolysis of virus-infected cells and tumor cells. Each of the NK cell populations has a unique receptor repertoire on its surface and, thus, unique functions. During their contact with a target cell, the most common mechanism of cytolysis is an exocytosis of lytic granules. However, some indirect evidence suggests that MV with CD56 phenotype and leukocyte-derived MV with various phenotypes are present in the peripheral blood plasma.This research is aimed to study the phenotype, composition and cytotoxic activity of microvesicles produced by NK cells. The analysis of receptor expression showed that MV, as well as source cells of the NK-92 cell line, had a similar CD56 molecule expression profile. The expression profile in MV differs from the same in source cells by higher CD119 and CD11b expression and by lower CD18 expression. Culturing of NK-92 cells in the presence of PMA, IL-1β, TNFα, IFNγ resulted in alterations of cell phenotypes and MV. Immunoblots revealed a change of perforin and granzyme B (GrB) in MV. The analysis of the cytotoxic activity of NK-92 cells in a natural killer in vitro assay employing K562 target cells demonstrated that MV obtained from TNFα-activated cells of the NK-92 cell line increased the cytotoxicity of the same TNFα-activated NK-92 cells regarding cytotoxicity levels. This coincides with the previously revealed increased content of GrB in MV obtained from TNFα-activated cells of the NK-92 cell line. To sum up depending on the cytokine NK-92 cells produce MV that differ in their phenotype, composition and activity. Any changes in MV composition can result in changes in their functional activity: in particular, changes can increase the cytotoxic activity of NK cells of the NK-92 cell line. Thus, besides a well-known and proved way for GrB delivery to a target cell, we can suggest an additional way – the transportation of GrB within MV.
    Type of Medium: Online Resource
    ISSN: 2313-741X , 1563-0625
    URL: Issue
    Language: Unknown
    Publisher: SPb RAACI
    Publication Date: 2019
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  • 2
    In: Medical Immunology (Russia), SPb RAACI, Vol. 22, No. 2 ( 2020-04-16), p. 249-268
    Type of Medium: Online Resource
    ISSN: 2313-741X , 1563-0625
    Language: Unknown
    Publisher: SPb RAACI
    Publication Date: 2020
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  • 3
    In: Medical Immunology (Russia), SPb RAACI, Vol. 23, No. 1 ( 2021-03-01), p. 95-106
    Abstract: Glaucoma is one of the leading causes of irreversible blindness worldwide, being an age-related disease. Its pathogenesis still is not fully understood. A particular interest is attracted to evaluation of the cytokine concentrations in the trabecular meshwork cell culture, and in the aqueous humor (AH) taken from the same patient, since such data may allow to describe more completely the glaucomatous trabecular changes and to clarify the mechanisms of intercellular interactions in pseudoexfoliative (PEX) glaucoma. The purpose of this study was a comparative analysis of cytokine contents in AH and in trabecular tissue (TT) supernatants in the patients with PEX glaucoma. The study included 23 eyes of patients with PEX glaucoma. The material studied was AH and supernatant of TT cell culture. The cytokine concentration was measured using a flow cytofluorimeter FacsCantoII (BD, USA) using the CBA method. SPSS version 19 software (IBM, USA) was used for the statistical data processing. Concentrations of cytokines (TNFα, IFNγ, IL-1β, IL-6, IL-8, IL- 10, VEGF, GM-CSF) were determined in AH and in the TT supernatant for each of the patients with PEX glaucoma. Only IL-6 and VEGF concentrations in AH were higher than those in the TT supernatant in patients with PEX glaucoma. The IL-6 concentration positively correlated with the VEGF and IL-8 concentrations in the TT supernatant. Correlations between other cytokines in the TT supernatant and AH were also identified and analyzed. Multiple regression analysis revealed that the duration of glaucoma and the IFNγ and TNFα concentrations in AH may have a significant influence on the corneal endothelial cells, being associated with density reduction in patients with PEX glaucoma. The correlation analysis did not reveal any links between other clinical data (corneal thickness in the optical center, IOP level, age) and the cytokine concentrations in the studied tissues. The obtained results suggest that only simultaneous analysis of the cytokine concentrations in the TT supernatant and AH taken from the same patient may provide a more complete description of the cytokine imbalance and pathological processes occurring in the trabecular meshwork in PEX glaucoma patients. It has been shown that the changing cytokine ratios observed in PEX glaucoma may be associated with development of uniform structural and functional changes in all tissues of the anterior eye segment.
    Type of Medium: Online Resource
    ISSN: 2313-741X , 1563-0625
    Language: Unknown
    Publisher: SPb RAACI
    Publication Date: 2021
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  • 4
    In: Russian Journal of Infection and Immunity, SPb RAACI, Vol. 8, No. 4 ( 2018-12-30), p. 511-
    Abstract: .
    Type of Medium: Online Resource
    ISSN: 2313-7398 , 2220-7619
    URL: Issue
    Language: Unknown
    Publisher: SPb RAACI
    Publication Date: 2018
    detail.hit.zdb_id: 3046274-5
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  • 5
    In: Medical Immunology (Russia), SPb RAACI, Vol. 24, No. 5 ( 2022-10-31), p. 1057-1064
    Abstract: Chronic psychosocial stress provokes anxious behavior and depressive disorders. The longitudinal stress-induced neuroendocrine signals may alter functioning of immune (central and peripheral) organs. Increased myelopoiesis is observed in bone marrow, being detrimental to lympho- and erythropoiesis, with increased emigration of monocytic bone marrow cells to the periphery and their acquisition of “inflammatory” phenotype. The subsequent migration of such monocytes to the brain with differentiation into the M1 type macrophages which form inflammatory signals, and their effect upon endothelial cells and microglia leads to increased production of cytokines, chemokines, and adhesion molecules, thus accelerating accumulation of bone marrow-derived monocytes migrating to the brain. The signals from bone marrow monocytes and activated microglia promote neuroinflammatory condition which leads to behavioral changes. Current data on the presence of non-resident bone marrow macrophages in the brain of depressed patients require studies of hematopoiesis in depression-like states. Pronounced plasticity is a characteristic feature of macrophages, i.e., their ability to acquire M1 or M2 phenotype depending on the microenvironment signals. M1 exhibit high pro-inflammatory activity and have neurodestructive properties, whereas M2 cells are characterized by low pro-inflammatory activity and pronounced regenerative potential, due to the production of multiple soluble mediators and cytokines, including neurotrophic and immunoregulatory factors, anti-inflammatory substances that provide neuroprotection, stimulate neurogenesis, synaptogenesis, growth and myelinization of axons, thus theoretically substantiating an opportunity of using the potential of M2 macrophages in the treatment of depression. In this work, we studied the effect of soluble factors of human macrophages, polarized into cells with M2 phenotype under the conditions of serum deprivation, upon bone marrow hematopoiesis and peripheral blood cells in a model of stress-induced depression. We have shown enhanced differentiation of hematopoietic stem cells into the granulocyte-macrophage (CFU-GM) lineage, along with increased monocyte population in peripheral blood in the depressive-like murine model. Development of a depressive-like state in the animals was associated with reduced amounts of both erythroid precursors in bone marrow and erythrocytes/hemoglobin in peripheral blood. Intranasal administration of soluble M2 macrophage factors (M2-SFs) for 7 days was accompanied by a corrective effect on the above parameters, being significant for peripheral blood monocytes. The data obtained suggest effectiveness of the M2-SFS anti-inflammatory effects in correcting changes in hematopoiesis caused by social stress in depressive-like animals.
    Type of Medium: Online Resource
    ISSN: 2313-741X , 1563-0625
    Language: Unknown
    Publisher: SPb RAACI
    Publication Date: 2022
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  • 6
    Online Resource
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    SPb RAACI ; 2021
    In:  Medical Immunology (Russia) Vol. 22, No. 6 ( 2021-01-10), p. 1111-1120
    In: Medical Immunology (Russia), SPb RAACI, Vol. 22, No. 6 ( 2021-01-10), p. 1111-1120
    Abstract: Our aim was to evaluate immunomodulatory properties of an original bioflavonoid complex in experimental immune disturbances induced by cyclophosphamide (Cy). We have studied morphometric indexes of thymus and spleen, as well as blood leukocyte counts, cell proliferative activity in lymphoid organs, delayed hypersensitivity responses to T cell-dependent antigen, along with differentiation activity of bone marrow stem cells in experimental animals during Cy-induced immune suppression after a course of bioflavonoid treatment. Suspension of the bioflafonoid complex was introduced to the male mice (СВАхC57Bl/6)F1 aged 12- 14 weeks at a daily dose of 2 mg/animal (80 mg/kg), per os, using gastric catheter, over 14 days. Cytostatic immunosuppression was produced by a single intraperitoneal Cy injection. Proliferative activity of spleen and thymic cells was determined by standard method with Н3 -thymidine incorporation in the 72-h cell culture. Cellular immune response was assayed by the degree of delayed-type hypersensitivity development in response to sheep erythrocytes. The number of hematopoietic progenitors was evaluated by culturing bone marrow cells in methylcellulose-based medium. The experiments have shown mitigation of immunosuppressive effects induced by Cy, in the course of bioflavonoid complex treatment, with respect to absolute and relative mass of lymphoid organs and leukocyte numbers in peripheral blood. Moreover, we have demonstrated decreased effects of Cy treatment upon the spontaneous activity of spleen cells, mitogen-induced thymocyte and splenocyte proliferation, intensivity of delayed-type hypersensitivity response that reached the values of intact animals. Following the course of bioflavonoids, we have revealed an increase in early hematopoietic progenitors. Alleviation of Cy-induced suppressive effects upon cellular immune response, proliferation rates of immune cells, as well as stimulation of hematopoietic stem cell functions suggest a sufficient capacity of the original bioflavonoid complex for modulation of immunity and hematopoiesis, thus presenting experimental proofs for its potential usage as an adjuvant treatment of the patients with malignant diseases.
    Type of Medium: Online Resource
    ISSN: 2313-741X , 1563-0625
    Language: Unknown
    Publisher: SPb RAACI
    Publication Date: 2021
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  • 7
    In: Medical Immunology (Russia), SPb RAACI, Vol. 21, No. 3 ( 2019-07-13), p. 427-440
    Abstract: Regulation of angiogenesis in the utero-placental bed determines adequate trophoblast invasion, placenta formation and development, as well as successful course of pregnancy. Natural killer (NK) cells, macrophages and trophoblast have the most significant effect on angiogenesis. To date, the functions of cells participating in placenta formation have been described in detail, both individually ( in vitr о ) and in tissues ( in situ ). However, no models have yet been created that reflect the interactions of NK cells, trophoblast and endothelium during angiogenesis. It remains unclear, how each cell population contributes to placental angiogenesis regulation, and to the cross-regulation of participating cell functions. Therefore, the aim of this research was to study contact and distant effects of NK cells upon formation of tube-like structures through co-culture of endothelial and trophoblast cells influenced by various cytokines (bFGF, VEGF, PlGF, TGF-β, IL-8, IFNγ and IL-1β). Introduction of NK cells to the co-culture of endothelial and trophoblast cells under conditions of both contact and distance-dependent culturing did not change the length of tube-like structures formed by endothelial cells. During contact-dependent culturing of NK cells with co-culture of endothelial and trophoblast cells in presence of IL-1β, the length of tubule-like structures remained unchanged, compared with the length of tube-like structures formed under the same culturing conditions, but without the cytokine added. During distant culturing of NK cells with co-culture of endothelial and trophoblast cells in the presence of IL-1β, the length of tube-like structures increased as compared with those formed under the same culturing conditions but without the cytokine. During contact-dependent (but not distant) culturing of NK cells with the co-culture of endothelial and trophoblast cells in the presence of VEGF, the length of tube-like structures was greater than those formed under the same culturing conditions but without the cytokine. When used in a three-component cell system, the pro-inflammatory cytokine IFNγhad no effect upon angiogenesis. During distant (but not contact-dependent) culturing of NK cells with co-culture of endothelial and trophoblast cells in the presence of TGF-β, the length of tube-like structures was less than the length of tube-like structures formed under the same culturing conditions but without the cytokine. Under conditions of distant culturing, TGF-βtriggered a signal in NK cells that inhibited angiogenesis. Decreased length of tube-like structures under conditions of a three-component cell co-culture in the presence of the following pro-angiogenic factors was revealed: IL-8, PlGF (during contact-dependent culturing only) and bFGF (during both contact-dependent and distant culturing). Thus, the effects of cytokines upon angiogenesis in a three-component co-culture (NK cells, trophoblast and endothelium) differed from those revealed previously in single-component (endothelium only) and two-component (co-culture of endothelium and trophoblast) cell models. The results of these experiments indicated that regulation of placental cell interactions involved both cellular contacts and effects produced by cytokines.
    Type of Medium: Online Resource
    ISSN: 2313-741X , 1563-0625
    URL: Issue
    Language: Unknown
    Publisher: SPb RAACI
    Publication Date: 2019
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  • 8
    In: Medical Immunology (Russia), SPb RAACI, Vol. 22, No. 4 ( 2020-08-07), p. 633-646
    Abstract: Extracellular vesicles that are shed from the plasma membrane contain a wide range of molecules, among  which  are proteins, lipids, nucleic  acids,  and sugars. The cytotoxic proteins of natural killer cells play a key role in the implementation of their cytolytic  functions. One of the important steps in understanding the distant  communication of cells is the determination of the proteome of microvesicles. This study was aimed at the protein profiling of the microvesicles produced by the NK-92 natural killer cell line. 986 proteins with a variety of functions were identified in the lysate of microvesicles using the MALDI-TOF mass spectrometric analysis.  With automated methods of functional analysis  applied, it has been  shown  that  the  largest  protein groups  are  hypothetical proteins, proteins with  unknown functions, and  domains. The  most  representative groups  are  also  comprised by  transcription  regulators; intracellular  signaling  proteins; RNA  translation, transcription, processing, and utilization regulators; receptors; protein processing  and proteolysis regulators; amino acid metabolism enzymes, as well as transport proteins and transport regulators. Minor functional groups are represented by vitamins and mineral metabolism enzymes, membrane and microdomain-forming proteins, hormones, hemostatic regulators, regulators of sensory  systems,  specific  mitochondrial and  Golgi  apparatus proteins, and extracellular signaling proteins. An intermediate position is occupied by various functional groups, including cytoskeleton and motor proteins; proteins of centrioles; ion channels and their regulators; proteins of the ubiquitin-proteasome pathway  of protein degradation; lipid,  steroid, and fatty acid metabolism enzymes; nucleic  acid  base and  carbohydrate metabolism enzymes, as well as energy  metabolism enzymes  and  other proteins involved  in intermediate metabolism; proteins of the immune response  and  inflammation; antigens and histocompatibility proteins; cytokines and growth factors; regulators of apoptosis, autophagy, endocytosis, and  exocytosis;  regulators of the  cell cycle and  division;  regulators of proliferation, cell differentiation, and morphogenesis; regulators of cell adhesion and  matrix  metabolism; nuclear transport proteins; transposition proteins; DNA  replication and  repair  proteins, as well as inactive  proteins. The  data  obtained expand  the existing knowledge of the distant  communication of cells and indicate new mechanisms of interaction between natural killer and target cells.
    Type of Medium: Online Resource
    ISSN: 2313-741X , 1563-0625
    Language: Unknown
    Publisher: SPb RAACI
    Publication Date: 2020
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  • 9
    In: Medical Immunology (Russia), SPb RAACI, Vol. 24, No. 2 ( 2022-04-20), p. 327-336
    Abstract: Endometriosis is a chronic gynecological disease, which poses a serious problem in terms of diagnosis and treatment. Despite decades of research, there are no specific signs and symptoms and no blood tests to clinically confirm the diagnosis, which makes timely diagnosis and treatment difficult. Therefore, the search for new markers for early non-invasive diagnosis of the disease remains relevant. Various subcellular structures involved in intercellular communication, in particular, microvesicles, can be considered promising biological markers for external genital endometriosis. The aim of this work was to assess the composition of microvesicles derived from leukocytes in the peripheral blood of patients with stage I-II of external genital endometriosis and the possibility of their use as markers of non-invasive diagnosis of peritoneal forms of endometriosis. The study involved 97 women aged 26-40 with stage I-II of external genital endometriosis, whose diagnosis was established intraoperatively and confirmed histologically. Pain syndrome was noted in all patients of the main group, with infertility also detected in 73.2% of the patients. The control group consisted of 20 patients, whose average age was 25.5±1.1 years, who were examined in connection with male infertility factor before the in vitro fertilization, and in whom, on the basis of intraoperative examination, presented no gynecological diseases, and no pain syndrome. Before the surgical intervention, peripheral blood was taken from all patients to determine the content of microvesicles derived from leukocytes. To isolate microvesicles, we used the previously described by M.P. Gelderman and J. Simak method. It was found that patients with stage I-II of external genital endometriosis experience an increase in the number of CD14 + , CD16 + and CD54 + CD14 + microvesicles in the peripheral blood by 1.1, 1.38 and 1.55 times, respectively, as well as a decrease in the number of CD45 + CD4 + , CD3 + CD4 + , CD3 + CD8 + microvesicles by 1.2, 4 and 1.5 times, respectively, compared with patients from the control group. Therefore, in patients with stage I-II of external genital endometriosis, an increase in the relative number of CD54 + CD14 + microvesicles in the peripheral blood above 5.22% can serve as a marker for early non-invasive diagnosis of the disease with sensitivity of 80.5% and specificity of 71%.  
    Type of Medium: Online Resource
    ISSN: 2313-741X , 1563-0625
    Language: Unknown
    Publisher: SPb RAACI
    Publication Date: 2022
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  • 10
    In: Medical Immunology (Russia), SPb RAACI, Vol. 23, No. 2 ( 2021-05-03), p. 275-292
    Abstract: Extracellular vesicles that are shed from the plasma membranes take an active part in intercellular communication, transporting a wide range of molecules, including proteins, lipids, nucleic acids and carbohydrates, being of great functional importance. One of the steps to better understanding of distant communications of cells and their regulatory mechanisms is a proteomic study of various extracellular vesicles, including microvesicles and exosomes. Pro-inflammatory cytokines produced by monocytes and individual complement system components play a key role in their specific functioning. The aim of this work was to study proteomic composition of THP-1 monocyte-like cells and their microvesicles. The MALDI-mass spectrometric analysis of electrophoretic protein fractions of cell lysates and microvesicles allowed for identifying 107 proteins that perform various functions. Among 19 determined functional groups, the largest ones comprise transcription regulators and proteins with unknown functions. The smallest functional groups include regulators of cell differentiation and development, proteins participating in immune response and inflammation, cellular receptors and their regulators, transporter and transport regulatory proteins, as well as cell proteins mediating adhesion and matrix structures, processing regulators, proteins of ubiquitin-proteasome system, intracellular signaling, autophagy and exocytosis regulators, chromatin structural proteins, hemostatic regulators, and peptide hormones. An intermediate position is occupied by cytokines and growth factors, enzymes, cytoskeleton and motor proteins, as well as RNA processing and translation regulators. The subsequent DAVID Functional Annotation Clustering analysis allowed for identifying the most common groups distributed by their molecular function, biological processes, and cellular component. Separately, in the microvesicles derived from THP-1 monocyte-like cells, proteins of the immune response and inflammation, cytokines and growth factors, intracellular signaling proteins, cell differentiation regulators and developmental proteins, as well as cell adhesion and matrix proteins were identified among other protein molecules. The data obtained on the partial proteome of THP-1 monocyte-like cells and their microvesicles extend the existing knowledge on distant communications between the cells and suggest new mechanisms of interaction between monocytes/macrophages and their microenvironment. 
    Type of Medium: Online Resource
    ISSN: 2313-741X , 1563-0625
    Language: Unknown
    Publisher: SPb RAACI
    Publication Date: 2021
    detail.hit.zdb_id: 3041761-2
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