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Umbilical cord blood as a promising source of NK cells for immunotherapy 脐带血作为一种有前途的免疫治疗NK细胞来源
Q4 Medicine Pub Date : 2023-06-01 DOI: 10.15789/1563-0625-ucb-2846
R. Velichinskii, J. Vavilova, A. Boyko, O. A. Shustova, A. Palamarchuk, G. Yusubalieva, O. N. Kucherova, M. Streltsova, E. Kovalenko
Currently, a large number of studies on genetic modification of cord blood NK cells (UCB-NK) are carried out at both clinical and preclinical levels. Immunotherapy based on UCB-NK cells has great potential for antitumor therapy. However, despite having known several advantages over peripheral blood NK cells (PB- NK), including a high concentration in cord blood and low virulence rate, UCB-NK cells are predominantly characterized in the scientific literature as immature and low-functioning NK cells. In this work, we studied the phenotypic characteristics of UCB-NK cells and the possibility of stimulatory compensation of the decreased functional activity of UCB-NK cells. Our studies revealed UCB-NK cells can be characterized as poorly differentiated and weakly activated cells with high level of inhibitory receptor NKG2A and low level of activating receptor NKG2C and HLA-DR, accordingly with the literature data. Two types of stimuli were chosen to stimulate freshly isolated UCB-NK cells: 1) 100 units of IL-2; 2) combinations of 100 units IL-2 and K-562 feeder cells expressing membrane-bound IL-21 (K562-mbIL21). It was shown the degranulation (LAMP-1) and proliferative activity was higher than for parallel cultured ex vivo PB-NK cells under the same conditions for UCB-NK cells stimulated for 7 days with IL-2 + K562-mbIL21. Moreover, stimulation in the way of IL-2 + K562-mbIL21 seemed to be a more perspective way to obtain a large number of proliferatively active UCB-NK cells compared to stimulation with IL-2 only. Since genetic modification of NK cells is a promising way to improve the antitumor properties of NK cells, retroviral transduction procedure was performed to study of the stimulated UCB-NK cells. UCB-NK cells stimulated with IL-2 + K562-mbIL21 were transduced on day 8 of cultivation. In this study, we used targeted overexpression of the adaptor molecule DAP12, which is involved in the signaling of activating NK cell receptors. PB-NK cells and UCB-NK cells were transduced under the equal experimental conditions in same volume of viral particles. As a result, the transduction efficiency was found to be more than 4-fold higher for UCB-NK cells compared to PB-NK cells. Thus, UCB-NK cells appear to be a promising tool for further research in cancer immunotherapy.
目前,大量关于脐带血NK细胞(UCB-NK)基因修饰的研究在临床和临床前水平均有开展。基于UCB-NK细胞的免疫治疗在抗肿瘤治疗中具有很大的潜力。然而,尽管已知与外周血NK细胞(PB- NK)相比有一些优势,包括脐带血中的高浓度和低毒力率,但在科学文献中,UCB-NK细胞主要以不成熟和低功能NK细胞为特征。在这项工作中,我们研究了UCB-NK细胞的表型特征以及UCB-NK细胞功能活性下降的刺激补偿的可能性。我们的研究表明,UCB-NK细胞具有低分化弱活化的特点,具有高水平的抑制性受体NKG2A,低水平的活化受体NKG2C和HLA-DR。选择两种类型的刺激刺激新分离的UCB-NK细胞:1)100单位的IL-2;2) 100个单位IL-2与表达膜结合IL-21 (K562-mbIL21)的K-562饲养细胞联合。结果表明,在相同条件下,IL-2 + K562-mbIL21刺激UCB-NK细胞7天后,其脱颗粒(LAMP-1)和增殖活性高于体外平行培养的PB-NK细胞。此外,与仅IL-2刺激相比,IL-2 + K562-mbIL21的刺激方式似乎是获得大量增殖活性UCB-NK细胞的更有前景的方式。由于NK细胞的遗传修饰是提高NK细胞抗肿瘤特性的一种有希望的方法,因此采用逆转录病毒转导程序来研究受刺激的UCB-NK细胞。IL-2 + K562-mbIL21刺激的UCB-NK细胞在培养第8天转导。在本研究中,我们使用靶向过表达adaptor分子DAP12,该分子参与激活NK细胞受体的信号传导。PB-NK细胞和UCB-NK细胞在相同的实验条件下,在相同体积的病毒颗粒中转导。结果,发现UCB-NK细胞的转导效率比PB-NK细胞高4倍以上。因此,UCB-NK细胞似乎是癌症免疫治疗进一步研究的一个有前途的工具。
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引用次数: 0
Associations of IL17A G-197A single nucleotide polymorphism with immunological parameters and structural changes of the brain in schizophrenia IL17A G-197A单核苷酸多态性与精神分裂症患者免疫参数和脑结构变化的关系
Q4 Medicine Pub Date : 2023-06-01 DOI: 10.15789/1563-0625-aoi-2806
I. Malashenkova, V. Ushakov, S. Krynskiy, D. Ogurtsov, N. Khailov, A. Ratushnyy, E. Filippova, N. Zakharova, G. P. Kostyuk, N. Didkovsky
Schizophrenia is a chronic mental disorder that is caused by a complex palette of genetic, epigenetic and environmental factors. Some of the important components of its pathogenesis are systemic inflammation and the dysfunction of immunity, which lead to neuroinflammation, contributing to development of structural brain changes. Earlier we have shown that increase in interleukin-17A levels is associated with morphometric changes and immune dysregulation in schizophrenia. IL17A G-197A (rs2275913) genetic polymorphism is involved in determining interleukin-17A secretion. The goal of this work was to investigate the associations between rs2275913 polymorphism, immune disorders and structural neurovisualization findings in schizophrenia to provide new insights into the immunopathogenesis of this disease. 60 patients aged 18 to 42 years diagnosed with schizophrenia were enrolled. 85 healthy volunteers were included into the control group. Multiplex assay was used to determine cytokine and chemokine serum levels. Rs2275913 polymorphism was assessed by polymerase chain reaction with electrophoretic detection of amplification products. A number of relationships between rs2275913 polymorphism and the immune parameters in schizophrenia were revealed. Carriers of G allele showed significant increase in IFNY, a key cytokine of Th1-link of adaptive immunity, and IL-8, an inflammatory chemokine. Also, increased levels of CXCL16 were observed in patients carrying the G allele. CXCL16 activates secretion of other proinflammatory chemokines and is involved in activation of Th1 adaptive immunity. Associations of heterozygous GA genotype with reduced cortical thickness in a number of areas of the frontal cortex in schizophrenia were found. Changes in cortical thickness in some of these areas, including middle frontal gyrus and orbitofrontal cortex, can be relevant to the pathogenesis of schizophrenia. The results highlight the importance of immunogenetic factors in the pathogenesis of schizophrenia and indicate that the rs2275913 polymorphism requires further studies as a potential biomarker of immune dysregulation and morphometric brain changes in schizophrenia.
精神分裂症是一种慢性精神障碍,由遗传、表观遗传和环境因素的复杂组合引起。其发病机制的一些重要组成部分是全身性炎症和免疫功能障碍,导致神经炎症,促进大脑结构变化的发展。早些时候,我们已经表明白细胞介素- 17a水平的增加与精神分裂症的形态变化和免疫失调有关。il - 17a G-197A (rs2275913)基因多态性与白细胞介素- 17a分泌有关。这项工作的目的是研究rs2275913多态性、免疫紊乱和精神分裂症结构神经可视化结果之间的关系,为这种疾病的免疫发病机制提供新的见解。60名年龄在18至42岁之间被诊断为精神分裂症的患者被纳入研究。85名健康志愿者作为对照组。多效法测定血清细胞因子和趋化因子水平。扩增产物电泳检测聚合酶链反应检测Rs2275913多态性。揭示了rs2275913多态性与精神分裂症患者免疫参数之间的一些关系。G等位基因携带者的IFNY(适应性免疫中th1通路的关键细胞因子)和IL-8(炎症趋化因子)显著升高。此外,在携带G等位基因的患者中观察到CXCL16水平升高。CXCL16激活其他促炎趋化因子的分泌,并参与Th1适应性免疫的激活。发现杂合子GA基因型与精神分裂症患者额叶皮质许多区域皮质厚度减少有关。其中一些区域的皮质厚度变化,包括中额回和眶额皮质,可能与精神分裂症的发病机制有关。这些结果强调了免疫遗传因素在精神分裂症发病机制中的重要性,并表明rs2275913多态性作为精神分裂症免疫失调和脑形态改变的潜在生物标志物需要进一步研究。
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引用次数: 0
Features of the immune status of patients with acute coronary syndrome who underwent СOVID-19, depending on the number of cytotoxic T lymphocytes (CD8+) 急性冠脉综合征患者接受СOVID-19治疗后的免疫状态特征:细胞毒性T淋巴细胞(CD8+)的数量
Q4 Medicine Pub Date : 2023-06-01 DOI: 10.15789/1563-0625-fot-2834
E. Safronova, L. V. Ryabova, A. Zurochka
The 2019 coronavirus disease (COVID-19) pandemic has had an unprecedented impact on health and economies around the world. Direct myocardial injury and cytokine storm, leading to destabilization of preexisting plaques and accelerated formation of new plaques, are two mechanisms that trigger the acute coronary syndrome in COVID-19. There is insufficient data on the immune status of patients with acute coronary syndrome who have undergone COVID-19. The aim of the study was to study T and B cell, humoral immunity depending on the number of cytotoxic T lymphocytes (CD8+) in patients with acute coronary syndrome who underwent COVID-19. Materials and methods of research: 65 men with unstable angina pectoris and acute myocardial infarction (acute coronary syndrome) from 40 to 65 years old, who had previously had COVID-19, were examined. A study of peripheral blood was carried out: complete blood count (Medonic device, Sweden), general and specific IgM, IgG, IgA, compliment fragments (Vector Best, Russia). Subpopulations of T and B lymphocytes were determined by flow cytometry. In persons with acute coronary syndrome who underwent COVID-19 with predominantly normal and elevated levels of cytotoxic T cells, a more severe course of the disease was observed: patients with acute myocardial infarction prevailed, they had longer mortality, longer treatment duration, and stent thrombosis was more common. In patients with elevated cytotoxic T cells, there was a maximum increase in erythrocytes, hemoglobin, hematocrit, lymphocytes of both the total number and subpopulations – T helpers, T-NK lymphocytes, NK lymphocytes, T lymphocytes of early and late activation, B1 and B2 lymphocytes, index of NBT-induced test. In patients with normal levels of NK cells, compared with other groups, there was an increase in spontaneous NBT activity and index, a significant decrease in C3a and C5a complement fragments. Prevalence of stent thrombosis and mortality in the group of patients with normal levels of cytotoxic T cells may indicate torpidity of the immune system in these patients with poor outcomes.
2019年冠状病毒病(COVID-19)大流行对世界各地的健康和经济产生了前所未有的影响。直接心肌损伤和细胞因子风暴,导致原有斑块的不稳定和新斑块的加速形成,是引发COVID-19急性冠状动脉综合征的两种机制。COVID-19感染的急性冠状动脉综合征患者的免疫状况数据不足。该研究的目的是研究T细胞和B细胞体液免疫依赖于细胞毒性T淋巴细胞(CD8+)数量的急性冠状动脉综合征患者感染COVID-19。研究材料和方法:选取40 ~ 65岁男性不稳定型心绞痛合并急性心肌梗死(急性冠状动脉综合征)患者65例,既往感染COVID-19。外周血研究:全血细胞计数(Medonic装置,瑞典),一般和特异性IgM, IgG, IgA,补体片段(Vector Best,俄罗斯)。流式细胞术检测T淋巴细胞亚群和B淋巴细胞亚群。在接受COVID-19的急性冠状动脉综合征患者中,细胞毒性T细胞水平主要正常和升高,观察到更严重的病程:急性心肌梗死患者普遍存在,他们的死亡率更长,治疗时间更长,支架血栓形成更常见。细胞毒性T细胞升高的患者红细胞、血红蛋白、红细胞压积、助T细胞总数和亚群淋巴细胞、T-NK淋巴细胞、NK淋巴细胞、早期和晚期活化T淋巴细胞、B1和B2淋巴细胞、nbt诱导试验指标均显著升高。在NK细胞水平正常的患者中,与其他组相比,自发NBT活性和指数增加,C3a和C5a补体片段明显减少。在细胞毒性T细胞水平正常的患者组中,支架内血栓形成的患病率和死亡率可能表明这些预后不良的患者的免疫系统处于迟钝状态。
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引用次数: 0
Metalloproteinase 9 and its role in the pathogenesis of allergic diseases in children 金属蛋白酶9及其在儿童变应性疾病发病中的作用
Q4 Medicine Pub Date : 2023-06-01 DOI: 10.15789/1563-0625-mai-2721
O. Semernik, A. Lebedenko, A. A. Appoeva
Metalloproteinases (MMP) play a significant role in the mechanisms of maintaining chronic inflammation and tissue remodeling. The study of concentration changes in these enzymes in the blood serum of children with allergopathology is of great practical and scientific interest.Objective: to study the role of MMP-9 in the pathogenesis of allergic diseases in children. 180 children aged from 1 to 18 years passed a comprehensive clinical and laboratory examination. This study included patients suffering from bronchial asthma (BA) (n = 54), atopic dermatitis (AD) (n = 54) and combination of these pathologies (n = 72). Serum levels of MMP9 were determined by enzyme immunoassay using Cloud-CloneCorp® test systems (USA).The analysis of the obtained data showed that among patients with the established diagnosis of BA, the maximum concentration of this cytokine was registered in children with a moderate course of the disease. The conducted correlation analysis showed the presence of a significant correlation between the severity of asthma and the level of control over the disease (r = 0.63). Similar data was obtained in patients with a combination of BA and AD. In children of this group, there was also a significant increase in serum MMP-9 compared with healthy patients (p = 0.015). The concentration of this matrix metalloproteinase in serum was slightly higher among children with polyvalent sensitization than in patients with monoallergic etiology of the disease (p = 0.272). The values of MMP-9 in patients with only skin manifestations of atopy were significantly higher than in the control group (p = 0.025).The data we obtained showed that all the patients we examined had a significant increase in the level of MMP-9 in the blood serum, which indicates an important role of this cytokine in the pathogenesis of allergic diseases in children.
金属蛋白酶(MMP)在维持慢性炎症和组织重塑的机制中发挥重要作用。研究过敏症患儿血清中这些酶的浓度变化具有重要的实用和科学意义。目的:探讨MMP-9在儿童变应性疾病发病机制中的作用。180名1至18岁的儿童通过了全面的临床和实验室检查。本研究纳入支气管哮喘(BA) (n = 54)、特应性皮炎(AD) (n = 54)以及这些疾病的合并(n = 72)患者。采用美国Cloud-CloneCorp®检测系统,采用酶免疫分析法测定血清MMP9水平。对所得数据的分析表明,在确定诊断为BA的患者中,该细胞因子浓度最高的是病程中等的儿童。相关分析显示哮喘严重程度与疾病控制水平之间存在显著相关性(r = 0.63)。在BA和AD合并的患者中也获得了类似的数据。与健康患者相比,该组患儿血清MMP-9水平也显著升高(p = 0.015)。多价致敏儿童血清中基质金属蛋白酶的浓度略高于单价致敏儿童(p = 0.272)。仅皮肤表现的特应性患者的MMP-9值显著高于对照组(p = 0.025)。我们获得的数据显示,我们检查的所有患者血清中MMP-9水平均显著升高,这表明该细胞因子在儿童变应性疾病的发病机制中起着重要作用。
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引用次数: 0
Effect of <i>Bifidobacterium bifidum</i> supernatant on the morphological and functional characteristics of human fibroblasts in real time during an <i>in vitro</i> experiment 两歧双歧杆菌的作用&lt;体外实时观察人成纤维细胞形态和功能特征的上清;实验
Q4 Medicine Pub Date : 2023-06-01 DOI: 10.15789/1563-0625-eob-2720
A. A. Markov, E. G. Kostolomova, T. Kh. Timokhina, G. S. Solovyev, Ya. I. Paromova, E. D. Polyanskih, K. A. Voronin
Currently, there is an active search for exogenous stimulators of repair and regeneration processes. In the recent decades, some data on the immunotropic activity of bifidobacteria have been accumulated. The key role in healing of wound defects belongs to fibroblasts due to the secretion of the extracellular matrix components, metabolites, signal factors for the surrounding cells, and tissue metabolism regulation. The paper presents the results of the study of the effect of Bifidobacterium bifidum supernatant (10 ml/mL) on the morphological and functional properties of human fibroblasts in real time during the in vitro experiment. In our work, we used the reference strain B. bifidum 791 (All-Russian Collection of Industrial Microorganisms of the State Research Institute for Genetics and Selection of Industrial Microorganisms “Genetika”, Deposit No. AS-1247) used in the production of the probiotic product “Bifidumbacterin” (ZAO “Ecopolis”, the city of Kovrov), and adult human fibroblasts (cell line LECH-4 (81)) (laboratory of cell cultures ENIIVI, the city of Yekaterinburg). Structural and functional studies were conducted on co-culture days 1, 3, 7, 14, 21, and 28. The products of B. bifidum secondary metabolism have a stressful effect on the morphological and functional state of fibroblasts on the first day. The processes of proliferation are stimulated in the culture in the experiment (2.67±0.24) compared with the control group (0.75±0.15) (p < 0.01) without blocking apoptosis in the cell. This leads to the increase in the production of extracellular matrix proteins, both collagen (pg/mL) (400±19 against 110±25 in the control group), and elastin (ng/mL) 395±30 and 125±29). Co-culture of fibroblasts within 24 hours in the experimental sample leads to a massive “release” of the CD44 receptor (p < 0.05), compared to the control group which is confirmed by phenotypic changes (r = 0.66). The decrease of CD105 + , CD44 + receptors (p < 0.05), compared with the control group and the increase of CD29 + expression (p < 0.05) is observed on days 1 and 3. Activated fibroblasts have an altered secretory phenotype that produces cytokines of various types such as TGF-b (r = 0.78), IL-6 (r = 0.57), IL-1b (r = 0.75), IL-8 (r = 0.63). The maximum adaptation of cells in the experimental system is registered on the 7th day, which correlates with morphometric (r = 0.59) and cytometric (r = 0.71) studies. The received data contribute to understanding of the mechanisms of the immunoregulatory influence of normal biota (in the bifidobacteria model) on the repair and regeneration processes.
目前,人们正在积极寻找修复和再生过程的外源性刺激因子。近几十年来,关于双歧杆菌的免疫活性已经积累了一些资料。成纤维细胞分泌细胞外基质成分、代谢物、周围细胞信号因子及组织代谢调节,在创面缺损愈合中起关键作用。本文介绍了两歧双歧杆菌上清液(10 ml/ ml)在体外实验中对人成纤维细胞形态和功能特性影响的实时研究结果。在我们的工作中,我们使用参考菌株双歧双歧杆菌791(全俄工业微生物收集,国家工业微生物遗传与选择研究所“Genetika”,存款号:AS-1247)用于生产益生菌产品“双歧杆菌”(ZAO“Ecopolis”,科夫罗夫市)和成人成纤维细胞(细胞系LECH-4(81))(叶卡捷琳堡市细胞培养实验室ENIIVI)。在共培养第1、3、7、14、21和28天进行结构和功能研究。双歧杆菌次生代谢产物对成纤维细胞在第一天的形态和功能状态有应激作用。与对照组(0.75±0.15)相比,实验组的增殖过程受到刺激(2.67±0.24)(p <0.01),不阻断细胞凋亡。这导致细胞外基质蛋白的产生增加,胶原蛋白(400±19 pg/mL,对照组为110±25 pg/mL)和弹性蛋白(395±30 ng/mL和125±29 ng/mL)。实验样品中24小时内的成纤维细胞共培养导致CD44受体的大量“释放”(p <0.05),与对照组相比,表型变化证实(r = 0.66)。CD105 +、CD44 +受体减少(p <0.05),与对照组相比,CD29 +表达升高(p <在第1天和第3天观察到0.05)。活化的成纤维细胞分泌表型改变,产生各种类型的细胞因子,如TGF-b (r = 0.78)、IL-6 (r = 0.57)、IL-1b (r = 0.75)、IL-8 (r = 0.63)。实验系统中细胞的最大适应性记录在第7天,这与形态学(r = 0.59)和细胞学(r = 0.71)研究相关。收到的数据有助于理解正常生物群(在双歧杆菌模型中)对修复和再生过程的免疫调节作用的机制。
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引用次数: 1
Quantification of C1 esterase inhibitor in human serum by enzyme-linked immunosorbent assay: Correlation with turbidimetric immunoassay 酶联免疫吸附法定量测定人血清中C1酯酶抑制剂:与比浊免疫法的相关性
Q4 Medicine Pub Date : 2023-06-01 DOI: 10.15789/1563-0625-qoc-2794
N. Gorbunov, A. Zhakhov, I. N. Gorbunova, A. M. Milichkina, I. Drozd, A. V. Gubanova, E. M. Danilova, R. N. Kuznecova, T. V. Savin, A. G. Burtseva, N. Pigareva, A. Ischenko, A. Totolian
C1 inhibitor of serine proteases (C1-INH) performs a regulatory function in the complement system and vascular permeability. Deficiency of C1-INH leads to various forms of angioedema, including hereditary angioedema (HAE). The cause of HAE is a genetically determined violation of the synthesis of C1-INH. A decrease in the level of C1-INH to 50% relative to the norm leads to an increase in the production of bradykinin, which is the basis for the diagnosis of HAE. The development of affordable ELISA for the quantitative determination of C1-INH is a popular direction for clinicians. During the development of a new kit for quantitative determination of C1-INH, two mouse monoclonal antibodies (mAb) with different epitope specificities were obtained. On their basis, a sandwich-type ELISA was developed. The specificity of the obtained mAb's was confirmed using the medical device “Berinert”. To prepare calibrators, C1-INH was affinity purified from human blood plasma using a sorbent with immobilized mAbs. The identity of the C1-INH protein was confirmed by PAGE electrophoresis, immunoblotting, and mass spectrometry on MALDI-TOF/TOF UltrafleXtreme mass spectrometer. To assess the quality indicators of developed reagents kit, studies were carried out in accordance with GOST R 51352-2013 and TU 21.20.23-041-01967164-2022. Values of quality indicators: accuracy — 93.53%; measurement linearity interval — 22.00-176.07 ng/mL. Using the developed ELISA test system, we examined 28 blood sera from healthy donors and 7 blood sera from patients with confirmed HAE. In the same samples, the content of C1-INH was determined by turbidimetric method, using the "Diagnostic reagents for in vitro immunochemical studies of specific blood proteins. Model: C1-esterase inhibitor (C1 EsteraseInhibitor)" (Aptec, Belgium). The correlation coefficient was 0.94 (p < 0.05). It was found that the diagnostic sensitivity and specificity of the developed ELISA is 100%. As a result of the study, an original ELISA test system for the quantitative determination of C1-INH was developed "Reagent kit for enzyme-linked immunosorbent assay of human C1-inhibitor (C1-inh PS)".
丝氨酸蛋白酶C1抑制剂(C1- inh)在补体系统和血管通透性中起调节作用。C1-INH缺乏导致各种形式的血管性水肿,包括遗传性血管性水肿(HAE)。HAE的病因是基因决定的C1-INH合成的破坏。C1-INH水平相对于正常值下降50%会导致缓激肽的产生增加,这是HAE诊断的基础。开发价格合理的酶联免疫吸附法(ELISA)用于C1-INH的定量检测是临床医生普遍关注的方向。在开发新的C1-INH定量检测试剂盒的过程中,获得了两种具有不同表位特异性的小鼠单克隆抗体(mAb)。在此基础上,建立了三明治型酶联免疫吸附试验。使用“Berinert”医疗器械确认获得的单抗的特异性。为了制备校准剂,C1-INH用固定化单抗吸附剂从人血浆中亲和纯化。通过PAGE电泳、免疫印迹和MALDI-TOF/TOF UltrafleXtreme质谱仪的质谱分析证实了C1-INH蛋白的身份。为评价所开发试剂试剂盒的质量指标,按照GOST R 51352-2013和TU 21.20.23-041-01967164-2022进行研究。质量指标值:准确度- 93.53%;测量线性区间- 22.00-176.07 ng/mL。使用开发的ELISA检测系统,我们检测了来自健康献血者的28份血清和来自确诊HAE患者的7份血清。在同一样品中,使用“特异性血液蛋白体外免疫化学研究诊断试剂”,用比浊法测定C1-INH的含量。型号:C1-酯酶抑制剂(C1 EsteraseInhibitor)(Aptec、比利时)。相关系数为0.94 (p < 0.05)。结果表明,所建立的ELISA诊断敏感性和特异性均为100%。本研究开发了一套用于C1-INH定量测定的独创ELISA检测系统“human C1-inhibitor (C1-INH PS)酶联免疫吸附测定试剂盒”。
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引用次数: 0
Evaluation of mediators of fibrosis and angiogenesis in the blood serum of premature infants with bronchopulmonary dysplasia 支气管肺发育不良早产儿血清纤维化和血管生成介质的评价
Q4 Medicine Pub Date : 2023-06-01 DOI: 10.15789/1563-0625-eom-2789
E. Semikina, M. Snovskaya, M. Basargina, A. A. Seliverstova, A. A. Zhuzhula, I. Davidova
In premature birth and postpartum damage to the developing lung, the processes of the formation of pulmonary vessels and alveoli are disrupted, leading to bronchopulmonary dysplasia (BPD). BPD is a multifactorial disease and the pathogenesis of lung tissue damage is still not fully understood. Studies of angiogenesis biomarkers can be informative for assessing the development of BPD. In this study we examined the blood serum of 65 premature infants aged 6 to 180 days of life; gestational age at birth was 23-33 weeks, body weight 480-1840 g, APGAR score 5-6. All children in the early neonatal period had respiratory distress syndrome, then 46 children formed and 19 did not form bronchopulmonary dysplasia. The concentration of the factors of angiogenesis and fibrosis was determined in blood serum by ELISA. There were no differences in the levels of angiopoietins 1 and 2, vascular endothelial growth factor VEGF-D, transforming growth factor beta TGF-β, thrombospondin-1. We observed a tendency to increasing the level of VEGF-A, which is a key regulator of angiogenesis and lung maturation; we regard this tendency as a favorable sign of lung formation. We found tendencies to increase of the adhesion molecule of endothelial platelet cells PECAM-1, interleukin 8 and connective tissue growth factor CTGF. CTGF expression is enhanced by artificial lung ventilation and exposure to high oxygen concentrations. We consider an increase of CTGF in BPD to be an unfavorable change, since the binding of CTGF to VEGF inhibits VEGF-induced angiogenesis. In children with BPD, we found a decrease in the level of platelet derived growth factor PDGF-BB, the median concentration was 3180 pg/mL in BPD versus 4782 pg/mL without BPD (p = 0.024). PDGF is an important factor in tissue regeneration and plays an important role in the formation of blood vessels. We assume the decreasing of PDGF concentration in BPD can lead to a violation of the alveolarization necessary for the formation of the structure of healthy lungs. Studies of angiogenesis factors will help to better understand the pathogenesis of lung damage in BPD.
在早产和产后肺发育损伤中,肺血管和肺泡的形成过程被破坏,导致支气管肺发育不良(BPD)。BPD是一种多因素疾病,其肺组织损伤的发病机制尚不完全清楚。血管生成生物标志物的研究可以为评估BPD的发展提供信息。在这项研究中,我们检测了65名6至180天早产儿的血清;出生胎龄23 ~ 33周,体重480 ~ 1840 g, APGAR评分5 ~ 6分。新生儿早期均出现呼吸窘迫综合征,46例形成,19例未形成支气管肺发育不良。采用ELISA法测定大鼠血清中血管生成因子和纤维化因子的浓度。两组间血管生成素1、2、血管内皮生长因子VEGF-D、转化生长因子TGF-β、血栓反应蛋白-1水平无显著差异。我们观察到VEGF-A水平增加的趋势,VEGF-A是血管生成和肺成熟的关键调节因子;我们认为这种倾向是肺形成的有利迹象。我们发现内皮血小板细胞粘附分子PECAM-1、白细胞介素8和结缔组织生长因子CTGF有增加的趋势。人工肺通气和暴露于高氧浓度下,CTGF表达增强。我们认为BPD中CTGF的增加是一种不利的变化,因为CTGF与VEGF结合会抑制VEGF诱导的血管生成。在患有BPD的儿童中,我们发现血小板衍生生长因子PDGF-BB水平下降,BPD患者的中位浓度为3180 pg/mL,而非BPD患者的中位浓度为4782 pg/mL (p = 0.024)。PDGF是组织再生的重要因子,在血管形成中起重要作用。我们认为BPD中PDGF浓度的降低可能导致健康肺结构形成所必需的肺泡化的破坏。血管生成因子的研究将有助于更好地了解BPD肺损伤的发病机制。
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引用次数: 0
Investigation of local expression of NLRP3 inflammasome complex genes in modeling retinal degeneration in vivo NLRP3炎性小体复合物基因在视网膜变性模型中的局部表达研究
Q4 Medicine Pub Date : 2023-06-01 DOI: 10.15789/1563-0625-iol-2780
N. V. Neroeva, O. A. Svitich, V. Neroev, A. R. Kinkulkina, N. Balatskaya, E. Sorozhkina
Neurodegenerative ophthalmopathology is one of the main causes of irreversible blindness and disability in the world. In the pathogenesis of diseases of this group, more and more attention has recently been paid to the role of local inflammation caused by the activation of innate immunity and the mechanisms of its genetic regulation. In recent years, works have appeared in the field of experimental ophthalmology that have demonstrated the possibility of NLRP1, NLRP3 inflammasome complexes assembling when exposed to hyperglycemia, oxygen deprivation of retinal cells, as well as modeling compressive stress similar to that in glaucoma [15]. However, the mechanism of inflammasome involvement in the development of neurodegenerative eye diseases remains unclear. The aim of the study was to investigate the local expression of genes encoding proteins of the NLRP3 inflammasome complex (NLRP3, CASP-1) in an experimental model of retinal degeneration in rabbits. The studies were performed on samples of tissue complex (TC) of the retina/retinal pigment epithelium (RPE) (retina/RPE TC), isolated from the eyes of 14 New Zealand albino rabbits, in which degenerative retinal lesion was modeled by a single subretinal injection of 0.01 mL of 0.9% sodium chloride solution, and 7 healthy rabbits without eye damage. The formation of retinal degeneration was judged on the basis of changes in morphofunctional parameters obtained during specialized ophthalmological research methods (optical coherence tomography, fundus autofluorescence, electroretinography) at follow-up periods of 1, 3 and 6 months. The level of expression of NLRP3 and CASP-1 genes in the retina/RPE TC was evaluated by reverse transcription polymerase chain reaction (RT-PCR). According to the results of the study, a statistically significant increase in NLRP3 gene expression (p < 0.001) was noted in the retina/RPE TC of experimental animals, which may indicate the involvement of NLRP-3 inflammasome components in the development of neurodegenerative retinal lesions. At the same time, the expression of the gene encoding CASP-1 was detected only in the retina/RPE TC of experimental eyes and is probably due to local inflammatory mechanisms in the retinal tissue.The high level of NLRP3, CASP-1 mRNA, detected in all retina/RPE TC samples of experimental eyes at late stages of the experiment (3 and 6 months), allows us to assume the formation of mechanisms (for example, activated glial phenotype) that support inflammation in retinal tissue. This should be taken into account in actively developing transplantation methods for the treatment of retinal degeneration.
神经退行性眼病理是世界上造成不可逆失明和残疾的主要原因之一。在这类疾病的发病机制中,近年来人们越来越关注先天免疫激活引起的局部炎症的作用及其基因调控机制。近年来,实验眼科学领域的研究表明,NLRP1、NLRP3炎性体复合物在暴露于高血糖、视网膜细胞缺氧以及模拟类似青光眼[15]的压应力时可能发生聚集。然而,炎性体参与神经退行性眼病发展的机制尚不清楚。本研究旨在探讨兔视网膜变性实验模型中编码NLRP3炎性小体复合物(NLRP3, CASP-1)蛋白的基因的局部表达。本研究取材于14只新西兰白化兔的视网膜/视网膜色素上皮(RPE)组织复合体(TC)(视网膜/RPE TC)样品,通过在视网膜下注射0.01 mL 0.9%氯化钠溶液建立退行性视网膜病变模型,同时取材于7只无眼部损伤的健康兔。在随访1、3、6个月期间,根据专业眼科研究方法(光学相干断层扫描、眼底自体荧光、视网膜电图)获得的形态功能参数的变化来判断视网膜变性的形成。逆转录聚合酶链反应(RT-PCR)检测NLRP3和CASP-1基因在视网膜/RPE TC中的表达水平。本研究结果显示,实验动物视网膜/RPE TC中NLRP3基因表达有统计学意义的升高(p < 0.001),这可能提示NLRP3炎性体成分参与了视网膜神经退行性病变的发生。同时,编码CASP-1的基因仅在实验眼的视网膜/RPE TC中检测到表达,可能与视网膜组织的局部炎症机制有关。在实验后期(3个月和6个月)的所有视网膜/RPE TC样本中检测到高水平的NLRP3, CASP-1 mRNA,使我们能够假设支持视网膜组织炎症的机制(例如,激活的胶质表型)的形成。在积极发展治疗视网膜变性的移植方法时应考虑到这一点。
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引用次数: 0
Hypoxia-induced factor-1α and markers of inflammation in patients with ischemic stroke 缺血性脑卒中患者缺氧诱导因子-1α与炎症标志物的关系
Q4 Medicine Pub Date : 2023-06-01 DOI: 10.15789/1563-0625-hif-2740
L. Pivovarova, I. Voznyuk, I. V. Osipova, O. Ariskina, E. A. Gogoleva, M. V. Prokhorova, N. G. Karpova
Ischemic stroke (IS) occurs as a result of local disturbance of hemocirculation and hypoxia in the brain tissue. Hypoxia-inducible factor-1α (HIF-1α), which is involved in the regulation of tissue oxygen levels, plays an important role in the pathophysiology of stroke, including neuronal survival, neuroinflammation, angiogenesis, glucose metabolism, blood-brain barrier permeability, and is important in IS outcomes. The purpose of the study was to determine the relationship between blood levels of HIF-1α and the degree of neurological deficit in the acute period of IS and the outcome of the disease. We examined 58 people with IS aged 73 (67-81) years. Patients were divided into two groups – discharged and dead. The severity of stroke (NIHSS), neurological deficit, comorbidity index, blood levels of HIF-1α, p53 protein, interleukin-6, cystatin C, CRP, creatinine, hematological parameters were determined at admission, on days 3 and 10 of the disease. At admission the blood levels of HIF-1α was lower than in the comparison group and remained reduced until the 10th day. On day 10 the association of HIF-1α with neurological deficit, comorbidity index and disease outcome was determined. We observed a feedback of HIF-1α with the content of erythrocytes, hemoglobin and hematocrit, which can be regarded as a reflection of the hemic component of mixed hypoxia. In dead patients, an increased blood level of cystatin C was detected, which was associated with HIF-1α concentrations. In all periods of observation of IS, a correlation between cystatin C and creatinine and CRP levels was noted. These results may indicate dysfunction of endotheliocytes, inflammation associated with hypoxia in IS. The prognostic significance of the blood level of HIF-1α on the 10th day for the outcome of IS was AUC = 0.900. Blood levels of HIF-1α in the acute period was associated with the severity of IS and the outcome of the disease.
缺血性中风是由于局部血液循环障碍和脑组织缺氧而发生的。缺氧诱导因子-1α (Hypoxia-inducible factor-1α, HIF-1α)参与组织氧水平的调节,在脑卒中的病理生理中发挥重要作用,包括神经元存活、神经炎症、血管生成、葡萄糖代谢、血脑屏障通透性,并在is预后中发挥重要作用。本研究的目的是确定IS急性期血液中HIF-1α水平与神经功能缺损程度及疾病结局之间的关系。我们检查了58例73岁(67-81)的IS患者。患者分为出院组和死亡组。测定患者入院时、发病第3天和第10天脑卒中严重程度(NIHSS)、神经功能缺损、合并症指数、HIF-1α、p53蛋白、白细胞介素-6、胱抑素C、CRP、肌酐、血液学参数。入院时血液中HIF-1α水平低于对照组,并持续降低至第10天。第10天测定HIF-1α与神经功能缺损、合并症指数和疾病结局的关系。我们观察到HIF-1α与红细胞、血红蛋白和红细胞压积的含量呈反馈关系,这可以看作是混合缺氧的血液成分的反映。在死亡患者中,检测到胱抑素C血药浓度升高,这与HIF-1α浓度有关。在观察IS的所有时期,胱抑素C、肌酐和CRP水平之间的相关性被注意到。这些结果可能表明内皮细胞功能障碍,炎症与缺氧相关。第10天血HIF-1α水平对IS预后的预测意义AUC = 0.900。急性期血液中HIF-1α水平与IS的严重程度和疾病结局相关。
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引用次数: 0
Diagnostically significant changes in subsets CD11b+CD64-CD32+CD16+, CD11b+CD64+CD32+CD16+ neutrophilic granulocytes of immunocompromised women with chronic infectious and inflammatory diseases of the genital tract of various etiologies 不同病因的生殖道慢性感染性和炎症性疾病免疫功能低下妇女CD11b+CD64-CD32+CD16+、CD11b+CD64+CD32+CD16+中性粒细胞亚群诊断意义的变化
Q4 Medicine Pub Date : 2023-06-01 DOI: 10.15789/1563-0625-dsc-2782
S. Kovaleva, I. Nesterova, S. N. Pikturno, E. I. Dydyshko, N. S. Prosolypova, А. M. Chulkova
Chronic pelvic inflammatory disease (PID) in women remains a problem due to the importance of medical consequences. The study of the receptor apparatus of neutrophilic granulocytes (NG) involved in anti-infective protection in diseases of various etiologies seems to be relevant. Aim: to clarify the features of variants of quantitative and phenotypic changes in subsets of NG CD11b+CD64-CD32+CD16+, CD11b+CD64+CD32+CD16+ of immunocompromised women during exacerbation of chronic PID of various etiologies. We were tested women 20-40 years: Study Group 1 (SG1, n = 20) – chronic PID during the exacerbation with mono- or mixed latent/recurrent various viral infections (chronic herpes-virus infections, papillomavirus infection, recurrent ARVI); Study Group 2 (SG2, n = 30) – chronic PID of bacterial etiologies; Comparison Group (CG)– 20 healthy women. The number of subsets CD11b+CD64-CD32+CD16+NG (major) and CD11b+CD64+CD32+CD16+NG (minor), receptor expression density (MFI) was determined (FC500, USA). It was found that in PID during the period of exacerbation, diagnostically significant differences in the subset composition of NG were revealed. We got a decrease in the CD11b+СD64-СD32+СD16+NG subset and in 7,6 times increase in the CD11b+CD64+CD32+CD16+NG subset in SG2 with chronic PID of bacterial etiology, in contrast to chronic PID occurring in combination with recurrent/persistent viral infection SG1. Negative transformation of NG subsets is associated with a predominant decrease in the level of expression of the activation CD16. The absence of an adequate response to the infectious and inflammatory process was revealed – the absence of an increase in the expression of the activation CD11b in the major subset in SG1, as well as in the minor subset in groups SG1 and SG2. In the major subset of NG in groups SG2 a decrease in the expression of the activation marker CD11b. In the various viral infections and PID (SG1), in the negatively altered minor subset of NG we got a decrease of expression of CD16, an increase of expression of CD64 and CD32. Determination of subsets of CD11b+СD64-СD32+СD16+, CD11b+CD64+CD32+CD16+NG and their phenotype can be used as diagnostic markers for the differential diagnosis of PID of viral and bacterial etiology, and for the development of new methods of targeted immunomodulatory therapy.
慢性盆腔炎(PID)在妇女仍然是一个问题,由于医疗后果的重要性。研究中性粒细胞(NG)受体装置参与各种病因疾病的抗感染保护似乎是相关的。目的:阐明不同病因的慢性PID加重期免疫功能低下女性NG CD11b+CD64-CD32+CD16+、CD11b+CD64+CD32+CD16+亚群定量和表型变化的变异特征。我们对20-40岁的女性进行了测试:研究组1 (SG1, n = 20) -慢性PID加重期间伴有单一或混合潜伏/复发性各种病毒感染(慢性疱疹病毒感染,乳头瘤病毒感染,复发性ARVI);研究2组(SG2, n = 30):细菌性慢性PID;对照组(CG): 20名健康女性。测定CD11b+CD64-CD32+CD16+NG(主要)和CD11b+CD64+CD32+CD16+NG(次要)亚群数量、受体表达密度(MFI) (FC500, USA)。结果发现,在加重期的PID中,NG的亚群组成在诊断上存在显著差异。我们发现,与合并复发/持续性病毒感染SG1的慢性PID相比,患有细菌性慢性PID的SG2中CD11b+СD64-СD32+СD16+NG亚群减少,CD11b+CD64+CD32+CD16+NG亚群增加7.6倍。NG亚群的负转化与活化CD16表达水平的显著下降有关。结果显示,对感染和炎症过程缺乏足够的反应——在SG1的主要亚群中,以及在SG1和SG2组的次要亚群中,活化CD11b的表达没有增加。在NG的主要亚群中,SG2组活化标记CD11b的表达减少。在各种病毒感染和PID (SG1)中,在NG负改变的次要亚群中,CD16的表达减少,CD64和CD32的表达增加。CD11b+СD64-СD32+СD16+、CD11b+CD64+CD32+CD16+NG亚群及其表型的测定可作为病毒性和细菌性PID鉴别诊断的诊断标志物,并为开发靶向免疫调节治疗的新方法提供依据。
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Medical Immunology (Russia)
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