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Role of minor lymphocyte populations in development of liver fibrosis in children with glycogen storage disease 小淋巴细胞群在糖原储存病儿童肝纤维化发展中的作用
Pub Date : 2023-08-11 DOI: 10.46235/1028-7221-10013-rom
O. Kurbatova, S. Petrichuk, G. Movsisyan, Daria G. Kuptsova, T. Radygina, A. Anushenko, E. Semikina, A. Potapov
Glycogen storage disease (GSD) is a rare condition that changes the way the body uses and stores glycogen. Objective: to evaluate the content of small populations of lymphocytes and their ratios in children with hepatic forms of glycogen disease depending on the stage of liver fibrosis. 148 children with GSD at the age of Me=7,7 [3,9;11,8] were examined. The comparison group consisted of 54 healthy children. The stage of liver fibrosis was carried out on the FibroScan F502 device (EchoSence, France). Immunophenotyping of lymphocytes was performed on CYTOMICS FC500 (Beckman Coulter, USA). Indicators of lymphocyte populations were analyzed as a percentage of deviation from the age norm. In children with GSD, an increase in the degree of liver fibrosis was revealed from age (R=0.57). Treg content in children with GSD was at the lower limit of the age norm and did not depend on the stage of liver fibrosis. The content of Th17 and Thact lymphocytes was significantly higher than that of the comparison group at all stages of liver fibrosis, starting from stage F1. With an increase in the stage of liver fibrosis, there was an increase in the proportion of patients with Thact and Th17 content exceeding the upper limit of the normative values (PF0-F4=0.021 and PF0-F4=0.012, respectively). An increase in Th17/Treg and Thact/Treg ratios was revealed in patients with GSD relative to the comparison group in all age groups, the dynamics of Th17/Treg and Thact/Treg ratios was characterized by their increase with age. Analysis of indicators depending on the stage of liver fibrosis in children with GSD revealed a significant increase in the Thact/Treg ratio from stage F0 to stages F1, F2, F3 and F4 (PF0-F4=0.000). The Th17/Treg index increased from stage F0 to stages F1, F2, F3 (PF0-F3=0.000). An increase in the content of Thact and Th17 lymphocytes, as well as Th17/Treg and Thact/Treg indices with an increase in the stage of liver fibrosis can be used as an additional tool in assessing fibrotic changes in the liver. Immunological indicators objectively reflect the severity of the patient's condition with hepatic forms of GSD.
糖原储存病(GSD)是一种罕见的疾病,它改变了身体使用和储存糖原的方式。目的:评价肝型糖原病患儿小群淋巴细胞的含量及其比例与肝纤维化分期的关系。对148例年龄为Me=7,7[3,9;11,8]的GSD患儿进行了检查。对照组为54名健康儿童。肝纤维化分期在FibroScan F502设备(echoense,法国)上进行。淋巴细胞免疫分型在美国Beckman Coulter公司的CYTOMICS FC500上进行。淋巴细胞群指标以偏离年龄标准的百分比进行分析。在GSD患儿中,肝纤维化程度随年龄增加而增加(R=0.57)。GSD患儿Treg含量处于年龄正常值的下限,且与肝纤维化分期无关。从F1期开始,肝纤维化各阶段Th17和Thact淋巴细胞含量均显著高于对照组。随着肝纤维化分期的增加,Thact和Th17含量超过正常值上限的患者比例增加(PF0-F4=0.021, PF0-F4=0.012)。各年龄组GSD患者Th17/Treg和Thact/Treg比值均较对照组升高,Th17/Treg和Thact/Treg比值随年龄的增长而升高。GSD患儿肝纤维化分期指标分析显示,从F0期到F1、F2、F3、F4期Thact/Treg比值显著升高(PF0-F4=0.000)。从F0期到F1、F2、F3期Th17/Treg指数升高(PF0-F3=0.000)。Thact和Th17淋巴细胞含量的增加,以及Th17/Treg和Thact/Treg指数随肝纤维化分期的增加,可作为评估肝脏纤维化变化的附加工具。免疫指标客观地反映了肝型GSD患者病情的严重程度。
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引用次数: 0
Effect of pregnancy-specific β1-glycoprotein on the expression of arginase-1 and indolamine-2,3-dioxygenase by myeloid-derived suppressor cells 妊娠特异性β1-糖蛋白对髓源性抑制细胞精氨酸酶-1和吲哚胺-2,3-双加氧酶表达的影响
Pub Date : 2023-08-11 DOI: 10.46235/1028-7221-10003-eot
V. Timganova, K. Shardina, E. Gutina
Myeloid-derived suppressor cells (MDSC) - a population of immature cells of myeloid origin with inhibitory functions, mainly related to T lymphocytes. Normally, MDSC account for less than 1% of leukocytes in peripheral blood. The number of these cells increases during healthy pregnancy. However, MDSC have been shown to play a critical role in the maintenance of tumor growth and in autoimmune diseases. Because MDSC are now considered important regulators of immunity, finding ways to manipulate their functions is important for the development of therapies for malignant and autoimmune diseases, as well as for pregnancy pathologies and post-transplant complications. The immunosuppressive mechanisms of these cells are mediated by their expression of the surface molecules CD73, ADAM17, PD -L1, the enzymes arginase 1 (Arg 1), inducible nitric oxide synthase (iNOS), and indoleamine 2,3-dioxygenase (IDO), reactive oxygen species, and the production of the anti-inflammatory cytokines IL -10 and TGF-1. Pregnancy-specific 1-glycoprotein (PSG) is a pregnancy glycoprotein that has immunomodulatory effects on natural (dendritic cells and macrophages) and adaptive (T cells) immunity cells. At the same time, the effect of PSG on MDSC has not been investigated so far. Since this glycoprotein has promising pharmacological applications, it is necessary to study not only the native variant of PSG but also its recombinant form. Since the main function of MDSC is immunosuppression, the aim of our work was to evaluate one of its mechanisms, namely the intracellular expression of amino acid degradation enzymes Arg1 and IDO under the influence of native and recombinant PSG in vitro. MDSC differentiation was performed from CD11b+ cells isolated from peripheral blood of healthy volunteers. Cells were cultured for 7 days with stepwise addition of GM-CSF, IL -1, and LPS. Native (n) (1, 10, and 100 g/mL) and recombinant (r) (1 and 10 g/mL) PSG was added to the cultures three days before the end of incubation. The percentage of MDSCs (Lin- HLA-DR -CD11b+CD33+) intracellularly expressing Arg1 and IDO was determined by flow cytometry. It was found that nPSG and rPSG did not alter the amount of Arg1-expressing MDSCs at all concentrations examined. However, at a concentration of 10 g/mL, both types of proteins caused a statistically significant increase in the percentage of cells expressing IDO. We have already established that nPSG and rPSG affect MDSC differentiation by increasing the proportion of these cells belonging to the monocytic subpopulation. However, now we can say that PSG, in addition, enhances the suppressive function of the studied cells. The obtained data are novel and open perspectives for targeting myeloid suppressor cells to improve cellular technologies in science and medicine.
髓源性抑制细胞(myeloid -derived suppressor cells, MDSC) -一群具有抑制功能的髓源性未成熟细胞,主要与T淋巴细胞有关。正常情况下,MDSC占外周血白细胞的比例不到1%。在健康怀孕期间,这些细胞的数量会增加。然而,MDSC已被证明在维持肿瘤生长和自身免疫性疾病中发挥关键作用。由于MDSC现在被认为是重要的免疫调节因子,因此找到操纵其功能的方法对于恶性和自身免疫性疾病以及妊娠病理和移植后并发症的治疗方法的发展非常重要。这些细胞的免疫抑制机制是通过其表面分子CD73、ADAM17、PD -L1、精氨酸酶1 (Arg 1)、诱导型一氧化氮合酶(iNOS)、吲哚胺2,3-双加氧酶(IDO)、活性氧的表达以及抗炎细胞因子IL -10和TGF-1的产生介导的。妊娠特异性1-糖蛋白(PSG)是一种对自然(树突状细胞和巨噬细胞)和适应性(T细胞)免疫细胞具有免疫调节作用的妊娠糖蛋白。同时,PSG对MDSC的影响目前还没有研究。由于该糖蛋白具有良好的药理应用前景,因此有必要不仅研究其天然变体,而且研究其重组形式。由于MDSC的主要功能是免疫抑制,我们的工作目的是评估其机制之一,即在体外天然和重组PSG的影响下,氨基酸降解酶Arg1和IDO在细胞内的表达。从健康志愿者外周血中分离的CD11b+细胞进行MDSC分化。细胞培养7天,逐步添加GM-CSF、IL -1和LPS。在培养结束前3天,将原生(n)(1、10和100 g/mL)和重组(r)(1和10 g/mL)的PSG加入培养物中。流式细胞术检测细胞内表达Arg1和IDO的MDSCs (Lin- HLA-DR - cd11b +CD33+)的百分比。结果发现,nPSG和rPSG在所有检测浓度下都没有改变表达arg1的MDSCs的数量。然而,在浓度为10 g/mL时,两种类型的蛋白质都引起表达IDO的细胞百分比的统计学显著增加。我们已经确定nPSG和rPSG通过增加这些属于单核细胞亚群的细胞的比例来影响MDSC分化。然而,现在我们可以说,PSG还增强了所研究细胞的抑制功能。所获得的数据为靶向骨髓抑制细胞以改善科学和医学中的细胞技术提供了新的和开放的视角。
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引用次数: 0
Disturbances in the b cell component of immune system and associated immune alterations in post-covid patients covid后患者免疫系统b细胞成分紊乱及相关免疫改变
Pub Date : 2023-08-11 DOI: 10.46235/1028-7221-9636-dit
M. Dobrynina, A. Zurochka, Mariia V. Komelkova, V. Zurochka, Alexey Sarapultsev
There is only limited data on B-cell response in post-COVID patients despite its importance in studying the post-infection immunity. The present study aimed to investigate the features of the B-cell response in post-COVID patients, focusing on various B cell phenotypes. Along with the standard immunogram, the following cell phenotypes were examined: common B cells (CD45+, CD3-, CD19+); common memory cells (CD45+, CD3-, CD19+, CD27+); common non-memory cells (CD45+, CD3-, CD19+, CD5+); B1 memory cells (CD45+, CD3-, CD19+, CD5+, CD27+); B2 memory cells (CD45+, CD3-, CD19+, CD5-, CD27+); B1 non-memory cells (CD45+, CD3-, CD19+, CD5+, CD27-); and B2 non-memory cells (CD45+, CD3-, CD19+, CD5-, CD27-). The study revealed a sharp increase in B1 memory cells in 15.3% of post-COVID patients with impaired levels of B1 memory cells. This increase was accompanied by elevated levels of total B memory cells, B1 total lymphocytes (mainly, B1 memory cells), total T lymphocytes, and total IgA. By contrast, the patients with impaired B1 memory cells exhibited a sharp decrease in plasma cells, B2 lymphocytes (both memory and non-memory cells), natural killer cells, T regulatory cells, early activation T cells (CD25+), and C3a complement fragment. These findings suggest a unique immune system disorder characterized by dysregulated B lymphocyte switching from IgM to IgG and IgA synthesis, thus resulting in marked decrease in B2 lymphocyte subpopulations. This disorder may be associated with reduced T regulatory lymphocytes and early activation of T lymphocytes responsible for regulating B lymphocyte differentiation. Furthermore, the patients also exhibited reduced hemoglobin and platelet parameters, thus, potentially, contributing to hypoxia and blood clotting abnormalities. Thus, the phenotype identification of these immune system disorders in post-COVID patients requires non-standard approaches to assessing immune status, thus compicating clinical examination, but highlighting the need for immunocorrective therapies. These findings contribute to better understanding of post-COVID immune system disorders and require further investigation into the underlying causal factors.
尽管b细胞反应对研究感染后免疫具有重要意义,但目前关于b细胞反应的数据有限。本研究旨在探讨covid后患者B细胞反应的特征,重点关注各种B细胞表型。与标准免疫图一起,检测了以下细胞表型:普通B细胞(CD45+, CD3-, CD19+);共同记忆细胞(CD45+、CD3-、CD19+、CD27+);常见非记忆细胞(CD45+、CD3-、CD19+、CD5+);B1记忆细胞(CD45+、CD3-、CD19+、CD5+、CD27+);B2记忆细胞(CD45+、CD3-、CD19+、CD5-、CD27+);B1非记忆细胞(CD45+、CD3-、CD19+、CD5+、CD27-);B2非记忆细胞(CD45+、CD3-、CD19+、CD5-、CD27-)。研究发现,在B1记忆细胞水平受损的患者中,15.3%的患者B1记忆细胞急剧增加。这种增加伴随着总B记忆细胞、B1总淋巴细胞(主要是B1记忆细胞)、总T淋巴细胞和总IgA水平的升高。相比之下,B1记忆细胞受损的患者浆细胞、B2淋巴细胞(包括记忆细胞和非记忆细胞)、自然杀伤细胞、T调节细胞、早期活化T细胞(CD25+)和C3a补体片段急剧减少。这些发现提示了一种独特的免疫系统紊乱,其特征是B淋巴细胞从IgM转向IgG和IgA合成的失调,从而导致B2淋巴细胞亚群的显著减少。这种疾病可能与T调节性淋巴细胞减少和负责调节B淋巴细胞分化的T淋巴细胞的早期激活有关。此外,患者还表现出血红蛋白和血小板参数降低,因此,可能导致缺氧和凝血异常。因此,covid - 19后患者中这些免疫系统疾病的表型鉴定需要非标准的方法来评估免疫状态,从而使临床检查复杂化,但突出了免疫矫正治疗的必要性。这些发现有助于更好地了解covid后免疫系统疾病,并需要进一步调查潜在的病因。
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引用次数: 0
Immunopharmacological aspects of studying immunotropic properties of a novel biocompound 研究一种新型生物化合物的免疫亲和性的免疫药理学方面
Pub Date : 2023-08-11 DOI: 10.46235/1028-7221-9643-iao
N. А. Zabokritskiy
The article presents an experimental study of immunotropic activity of a new compound, a metabiotic, based on metabolites (biologically active substances) produced by a of saprophytic Bacillus subtilis B-9909 strain, being safe for health and standardized at Russian National Collection of Industrial Microorganisms (RNCIM). The aim of our study was to experimentally evaluate the immunotropic effects of metabolites produced by the probiotic Bacillus subtilis B-9909 upon the parameters of cellular immunity tested in the animal model of induced toxic liver damage. The metabolites were isolated from the cultural liquid of Bacillus subtilis culture (RNCIM strain B-9909) during its deep cultivation in a medium containing hydrochloric acid hydrolyzate of soy flour, or pancreatic hydrolyzate of casein. At 16-18 hours of cultivation, the cells were in the exponential growth phase. The indices of cellular status were studied in experimental groups of animals, when assessing therapeutic efficacy of an experimental metabiotic sample, as compared to a group of laboratory animals treated with the reference drug (ursosan). We assessed the quantitative indicators of blood as follows: determination of phagocytic activity (FA) of peripheral blood neutrophils; measurement of metabolic activity in peripheral blood neutrophils by means of NBT-test; quantitative determination of T and B lymphocytes; the number of antibody-forming cells (AFC). Liver damage was studied in a model of acute toxic hepatitis in albino rats. Experimental toxic hepatitis was induced by 40% solution of CCl4 in vaseline oil injected intragastrically for 2 weeks at the dose of 0.2 g/kg. The results of experimental studies showed activation of phagocytic activity by peripheral blood neutrophils at the early terms of experiment, being also confirmed by the results of NBT-testing. Moreover, at the early observation terms, a significantly increased representation of T and B lymphocyte populations as well as AFC numbers, were revealed thus suggesting activation of all components of cellular immunity in response to the carbon tetrachloride toxicity. Thus, the reported studies on the cellular status of laboratory animals treated with metabolites produced by probiotic microorganisms of the genus Bacillus subtilis (B-9909) on laboratory animals with toxic liver damage allow us to conclude that the test sample of this metabiotic preparation exerts a significant immunomodulatory effect, when compared with ursosan. This evidence allows us to consider this compound a promising candidate drug for a new hepatoprotector with immunotropic effect.
本文对一种经俄罗斯国家工业微生物收集(RNCIM)标准化的腐生枯草芽孢杆菌B-9909菌株的代谢物(生物活性物质)进行了免疫活性的实验研究。本研究旨在通过实验研究枯草芽孢杆菌B-9909代谢产物对中毒性肝损伤动物模型细胞免疫指标的免疫效应。这些代谢物是从枯草芽孢杆菌(RNCIM菌株B-9909)在含有大豆粉盐酸水解物或酪蛋白胰腺水解物的培养基中深度培养的培养液中分离出来的。培养16 ~ 18 h时,细胞进入指数生长期。在评估实验代谢样品的治疗效果时,对实验组动物的细胞状态指标进行了研究,并与使用参比药物(熊糖)治疗的一组实验动物进行了比较。我们评估血液的定量指标如下:外周血中性粒细胞吞噬活性(FA)的测定;用nbt法测定外周血中性粒细胞代谢活性;T、B淋巴细胞的定量测定;抗体形成细胞(AFC)的数量。用急性中毒性肝炎模型研究了白化大鼠肝损伤。采用凡士林油中40% CCl4溶液,以0.2 g/kg剂量灌胃2周,致实验性中毒性肝炎。实验研究结果表明,外周血中性粒细胞在实验初期就激活了吞噬活性,nbt检测结果也证实了这一点。此外,在早期观察期内,T和B淋巴细胞群以及AFC数量的代表性显著增加,这表明细胞免疫的所有组成部分都被激活,以响应四氯化碳毒性。因此,有报道称,枯草芽孢杆菌属益生菌(B-9909)产生的代谢物对中毒性肝损伤实验动物细胞状态的研究,使我们得出结论,与熊熊糖相比,这种代谢物的测试样品具有显著的免疫调节作用。这一证据使我们认为该化合物是一种具有免疫效应的新型肝保护剂的有希望的候选药物。
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引用次数: 0
Features of T cell immunity depending on the content of natural killer cells in patients with acute coronary syndrome following COVID-19 COVID-19后急性冠状动脉综合征患者T细胞免疫特征与自然杀伤细胞含量的关系
Pub Date : 2023-08-11 DOI: 10.46235/1028-7221-9640-fot
E. Safronova, L. V. Ryabova
We examined 65 men with unstable angina and acute myocardial infarction (acute coronary syndrome - ACS) from 40 to 65 years old who had previously had COVID-19 and 20 people with ACS who had not undergone COVID-19. All persons also had hypertension, and they required stenting of the coronary arteries within the next 3 days after admission to the hospital. From the immunological parameters by flow cytometry on the Navios cytofluorimeter (BeckmanCoulter, USA), according to the standardized technology for assessing the lymphocytic link of immunity [1], the following were determined: ), CD45+ CD3+ CD8+ (cytotoxic T-lymphocytes), CD45+CD3-CD19+ (B-lymphocytes), CD45+CD3+CD16+CD56+ (TNK cells), CD45+CD3-CD16+CD56+ (natural killer cells), CD45+ CD3+CD4+CD25+CD127- (T-regulatory cells), CD45+ CD3+CD4+CD25+ (T-lymphocytes - early activation), CD45+CD3+HLA-DR (T-lymphocytes - late activation). All patients were divided into groups depending on the content of NK cells (natural killer cells). Patients who have had COVID-19 have 3 phenotypes of disorders (decreased NK cell count, normal and increased), while non-survivors have 2 phenotypes (decreased NK cell count and normal). The most severe condition and severity of immune disorders were found in patients who had undergone COVID-19. In patients with acute coronary syndrome and COVID-19, predominantly with normal and elevated levels of NK cells, compared with ACS patients without COVID-19, a more severe course of the disease was observed - patients with acute myocardial infarction prevailed, they had a higher mortality rate, the duration of treatment was increased, and stent thrombosis was also more common. In persons with ACS and COVID-19 with elevated NK cells, the maximum decrease in the T-cell immunity was observed: T-lymphocytes of general, T-lymphocytes-helpers, T-cytotoxic lymphocytes, T-lymphocytes of early activation, T-regulatory cells in absolute numbers compared to other groups. The lowest immunoregulatory index and, at the same time, the maximum number of T-NK-lymphocytes were observed in persons who had undergone COVID-19 and had reduced NK cells. The minimum number of T-NK lymphocytes was recorded in patients with low NK cells who did not have COVID-19. Minimal T-lymphocytes (CD45+CD3+CD4+HLA-DR+) of late activation were found in people who recovered from COVID-19 with elevated and normal NK cells. The lowest number of late activation regulatory T cells was observed in patients who did not have COVID-19, but were vaccinated, and had a normal content of NK cells. The study also allows us to more clearly define the groups of patients with ACS who need additional immunocorrection.
我们检查了65名患有不稳定心绞痛和急性心肌梗死(急性冠状动脉综合征- ACS)的男性,年龄在40至65岁之间,之前患有COVID-19, 20名患有ACS但未经历COVID-19。所有患者均患有高血压,并在入院后3天内需要行冠状动脉支架置入。从Navios细胞荧光仪(BeckmanCoulter, USA)流式细胞术的免疫学参数来看,根据评估免疫淋巴细胞环节的标准化技术[1],确定如下:)、CD45+CD3+ CD8+(细胞毒性t淋巴细胞)、CD45+CD3+ cd19 + (b淋巴细胞)、CD45+CD3+CD16+CD56+ (TNK细胞)、CD45+CD3-CD16+CD56+(自然杀伤细胞)、CD45+CD3+ CD4+CD25+CD127- (t调节细胞)、CD45+CD3+ CD4+CD25+ CD25+ (t淋巴细胞-早期活化)、CD45+CD3+ CD4+CD25+ (t淋巴细胞-晚期活化)。所有患者根据NK细胞(自然杀伤细胞)的含量进行分组。感染COVID-19的患者有3种表型(NK细胞计数减少、正常和增加),而非幸存者有2种表型(NK细胞计数减少和正常)。感染COVID-19的患者病情最严重,免疫功能紊乱程度最高。在急性冠状动脉综合征合并COVID-19的患者中,NK细胞水平以正常和升高为主,与未合并COVID-19的ACS患者相比,病程更为严重——以急性心肌梗死患者为主,死亡率更高,治疗时间更长,支架内血栓形成也更为常见。在ACS和COVID-19合并NK细胞升高的患者中,t细胞免疫功能的下降幅度最大:一般t淋巴细胞、辅助t淋巴细胞、毒性t淋巴细胞、早期活化t淋巴细胞、调节性t淋巴细胞的绝对数量与其他组相比。在感染COVID-19且NK细胞减少的人群中,免疫调节指数最低,同时t -NK淋巴细胞数量最多。无COVID-19的低NK细胞患者记录了T-NK淋巴细胞的最低数量。在NK细胞升高和正常的COVID-19康复患者中发现了最小的晚期激活t淋巴细胞(CD45+CD3+CD4+HLA-DR+)。未感染COVID-19但接种疫苗且NK细胞含量正常的患者中,晚期活化调节性T细胞数量最少。该研究还使我们能够更清楚地定义需要额外免疫矫正的ACS患者群体。
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引用次数: 0
The effectiveness of immunomodulatory therapy with a synthetic analogue of the active center of the hormone thymus thymopoietin in the complex treatment of immunocompromised women with chronic infectious and inflammatory diseases of the pelvic organs 胸腺激素活性中枢的合成类似物免疫调节疗法在盆腔器官慢性感染性和炎症性疾病免疫功能低下妇女的综合治疗中的有效性
Pub Date : 2023-08-11 DOI: 10.46235/1028-7221-10011-eoi
S. Kovaleva, I. Nesterova, S. N. Pikturno, G. Chudilova, L. Lomtatidze, Yu. V. Teterin, A.I. Pirogova, N. S. Prosolypova, А. M. Chulkova
Chronic inflammatory diseases of the pelvic organs (pelvic inflammatory disease, PID) in women is among the main understudied problems in gynecology worldwide with adverse medical and socio-economic consequences, thus justifying the need for further study of immunopathogenesis and development of new approaches to treatment. Our objective was to develop new immunotherapeutic approaches to correction of combined disorders of the immune system functioning in immunocompromised women with PID and to evaluate their clinical and immunological efficacy. 55 women aged 20-40 years were examined, i.e., 35 patients with exacerbation of sluggish or recurrent PID, resistant to conventional therapy. Testiung was performed before complex treatment (study group 1 GI- 1) and after the course (study group 2 GI-2). Contents of T and B lymphocytes, natural killer cells (NK) (CYTOMICS FC500, USA), phagocytic and microbicidal functions of neutrophilic granulocytes (NG) were assessed in SG-1 and SG-2 before and 2-3 days after complex treatment with addition of an immunotherapeutic drug based on hexapeptide (HP) at a daily dose of 45 mg/ml intramuscularly for 10 days. In patients from SG-1, a decrease in T cells (CD3+CD19- ) and B cells (CD3-CD19+), a 2-fold increase in the content of NK CD3-CD16+CD56+ was found, along with altered functioning of NG (deficiency of actively phagocytizing NG, a decrease in their digestive function and NADPH-oxidase activity). In SG-2 patients, the treatment was followed by restoration of the T (CD3+CD19-) and B cells (CD3-CD19+), NK cells (CD3-CD16+CD56+), like as an increase in effector functions, i.e., microbial capture by NG and their killing ability due to activation of NADPH oxidases and normalization of microbicidal reserve capacity in the NG cell population. Positive clinical effect included reduction of clinical symptoms in acute period, absence of PID exacerbations over follow-up for 6 months (85.6% of cases). Occasional exacerbations of PID were associated with medical manipulations (5.7%) and unprotected sexual contacts (5.7%). The immunopathogenetically proven approach to correction of combined functional impairment of immune system in the women with PID shows a positive clinical and immunological effect.
女性盆腔器官慢性炎症性疾病(盆腔炎,PID)是世界范围内妇科研究不足的主要问题之一,具有不良的医疗和社会经济后果,因此有必要进一步研究免疫发病机制和开发新的治疗方法。我们的目标是开发新的免疫治疗方法来纠正免疫功能低下的PID妇女的免疫系统功能紊乱,并评估其临床和免疫学疗效。55名年龄在20-40岁的女性,即35例迟滞或复发性PID加重,常规治疗无效。在综合治疗前(研究组1 GI- 1)和疗程后(研究组2 GI-2)分别进行测试。以六肽免疫治疗药物(HP)为基础,每日45 mg/ml肌注治疗10天,观察SG-1和SG-2在治疗前和治疗后2-3天的T淋巴细胞和B淋巴细胞、自然杀伤细胞(NK)(美国CYTOMICS FC500)的含量,以及嗜中性粒细胞(NG)的吞噬和杀微生物功能。在SG-1患者中,发现T细胞(CD3+CD19-)和B细胞(CD3-CD19+)减少,NK CD3- cd16 +CD56+含量增加2倍,同时NG功能改变(活性吞噬NG缺乏,消化功能和nadph氧化酶活性降低)。在SG-2患者中,治疗后恢复T细胞(CD3+CD19-)和B细胞(CD3-CD19+), NK细胞(CD3- cd16 +CD56+),如效应功能的增加,即NG捕获微生物及其由于激活NADPH氧化酶和NG细胞群中杀微生物储备能力的正常化而产生的杀伤能力。临床疗效包括急性期临床症状减轻,随访6个月无PID加重(85.6%)。PID的偶尔加重与医疗操作(5.7%)和无保护的性接触(5.7%)有关。经免疫病理学证实的治疗PID女性合并免疫系统功能障碍的方法具有积极的临床和免疫学效果。
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引用次数: 0
Indicators of immunological response to non-drug therapy of chronic adenoiditis 慢性腺样体炎非药物治疗的免疫反应指标
Pub Date : 2023-08-11 DOI: 10.46235/1028-7221-9448-ioi
A. V. Solodovnic, T. A. Vyalova, E. V. Gabova, Pavel A. Pozdnyakov, Daria S. Bushueva
The aim of our work was to study individual indices of immune response in children with chronic adenoiditis and evaluate their dynamics following low-frequency ultrasonic cavitation combined with photochromotherapy included into the treatment regimen. The study involved 104 patients 3 to 15 years old, divided into three groups: the 1st control group (n = 34) received standard treatment for 7 days; 2nd group (n = 37) received a supplementary low-frequency ultrasonic cavitation irrigation of pharyngeal tonsil combined with photochromotherapy on lymphoid tissue of the pharyngeal tonsil for 7 days; the 3rd group (n = 33) received only low-frequency ultrasonic cavitation irrigation treatment. Comparative estimation of clinical and immunological indicators (sIgA, IL-1, IL-6, IL-8, IL-10, TNF) was performed prior to the therapy (day 0) and on the 7th day from the start of treatment. Before therapy, a decrease in the content of sIgA, IgA was revealed in all groups. In the second group, there was a statistically significant increase in the level of IgA after treatment, which suggests activation of local immunity factors. The dynamics of proinflammatory cytokines (IL-6, IL- 10) also indicates effectiveness of the drug treatment by reducing manifestations of the tissue immunological reactivity in pharyngeal tonsil. An increased number of anti-inflammatory IL-8 and IL- 10 cytokines could be considered a compensatory response to decreased level of proi-inflammatory cytokines. As a result, the inclusion of low-frequency ultrasonic cavitation in combination with photochromotherapy into the complex treatment for chronic adenoiditis thus stabilizing the course of immune response.
本研究的目的是研究慢性腺样体炎儿童在低频超声空化联合光显色治疗后免疫反应的个体指标,并评估其动态变化。研究纳入104例3 ~ 15岁的患者,分为三组:第一对照组(n = 34)接受标准治疗7天;第二组(37例)给予咽扁桃体辅助低频超声空化灌洗联合咽扁桃体淋巴组织光显治疗7 d;第三组(n = 33)只接受低频超声空化灌洗治疗。在治疗前(第0天)和治疗开始后第7天进行临床和免疫学指标(sIgA、IL-1、IL-6、IL-8、IL-10、TNF)的比较评估。治疗前各组sIgA、IgA含量均下降。第二组治疗后IgA水平升高,有统计学意义,提示局部免疫因子激活。促炎细胞因子(IL-6、IL- 10)的动态变化也通过降低咽扁桃体组织免疫反应性的表现表明药物治疗的有效性。抗炎IL-8和IL- 10细胞因子数量的增加可以被认为是对促炎细胞因子水平下降的代偿反应。因此,将低频超声空化联合光显色疗法纳入慢性腺样体炎的综合治疗中,从而稳定了免疫反应的过程。
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引用次数: 0
Studies on the hormone and cytokine producing function of human cumulus cells and its interrelation with fertility in polycystic ovarian syndrome 多囊卵巢综合征人积云细胞激素和细胞因子分泌功能及其与生育关系的研究
Pub Date : 2023-08-11 DOI: 10.46235/1028-7221-9999-sot
O. V. Valikova, V. Zdor, Yakov N. Tikhonov, A. Boroda, A. M. Gracheva, K. G. Kolbin
Cumulus cells (CC) are the specialized layer of follicular cells that are in close contact with the oocyte. They are considered as indirect markers of the oocyte quality. Changes of these cells suggest a damage of the ovary. Determination of cytokines in cumulus cell culture may predict the chance for the conception and development of pregnancy. The objective of the present study was to obtain a primary culture of CC from healthy donors and patients with polycystic ovarian syndrome (PCOS), to identify the most significant differences in production of key cytokines in the CC monocultures of patients and healthy individuals in order to predict the results of in vitro fertilization. Materials and methods: the cell culture technique was used, i.e., cumulus cells of healthy donors and patients with polycystic ovary syndrome were obtained by transvaginal puncture of follicles in the in vitro fertilization (IVF) program. This procedure does not affect the rights of embryo, since the CC are not used at the stage of IVF procedure and intracytoplasmic sperm injection (Ethical Committee Protocol No. 9 of May 16, 2022). IL-6, IL-10, IFN and progesteroneparameters were tested in adhesive cultures of CC by ELISA technique on days 1, 3, 7 of in vitro experiments. Results: We revealed a continuous secretion of progesterone, IL-6, IL-10, IFN in adhesive monocultures of CC. In the patients with PCOS, we have found a sharp increase of progesterone level in cultural media (p 0.01) on the 7th day, By contrast, the initially increased progesterone levels proved to be significantly decreased in donors on the 7th day of culture. Moreover, in the culture of CC from patients with PCOS (7th day of the experiment), the values of IL-6, IL-10 increased only two-fold compared with 30-fold increase of these cytokines in healthy donors (p 0.01). At the same term, we have observed a threefold decrease in IFN in the CC cultures of PCOS patients (p1-7 0.01; p 0.05) compared with cultured controls, which showed a 20-fold increase (p1-7 0.01), thus determining differences in total cytokine balance and, probably, influencing the pregnancy prognosis. Conclusion: Significant multidirectional changes of cytokine levels in the culture of cumulus cells of the patients with PCOS and in healthy individuals may be regarded as determining factors in formation of blastocyst and preservation of the embryo. A further in vitro research on the production of cytokines and sex steroids by CC is especially important on day 5 to 7, when the oocytes are selected for entry into the in vitro fertilization cycle. The study of morpho-functional properties of little-studied cumulus cells using the cell culture technique will enable us for a deeper study on the mechanisms of disturbed folliculogenesis in PCOS, and, thereby, improve the reproductive prognosis in this disorder.
积云细胞(CC)是卵泡细胞的特化层,与卵母细胞密切接触。它们被认为是卵母细胞质量的间接标志。这些细胞的变化表明卵巢受损。积云细胞培养中细胞因子的测定可以预测妊娠的受孕和发育机会。本研究的目的是获得健康供体和多囊卵巢综合征(PCOS)患者的CC原代培养,以确定患者和健康个体的CC单培养中关键细胞因子产生的最显著差异,从而预测体外受精的结果。材料和方法:采用细胞培养技术,即在体外受精(IVF)方案中经阴道穿刺卵泡获得健康供体和多囊卵巢综合征患者的积云细胞。该程序不影响胚胎的权利,因为CC不用于体外受精程序和卵胞浆内精子注射阶段(伦理委员会2022年5月16日第9号议定书)。在体外实验的第1、3、7天,采用ELISA技术检测CC黏附培养物中IL-6、IL-10、IFN和孕酮参数。结果:单核细胞黏附培养液中持续分泌黄体酮、IL-6、IL-10和IFN, PCOS患者培养液中黄体酮水平在培养第7天急剧升高(p < 0.01),而供体的黄体酮水平在培养第7天开始升高,而供体的黄体酮水平明显下降。此外,在PCOS患者CC的培养中(实验第7天),IL-6、IL-10的值仅增加了2倍,而健康供者的这些细胞因子的值增加了30倍(p 0.01)。在同一时期,我们观察到PCOS患者CC培养物中IFN下降了三倍(p1-7 0.01;P < 0.05),比培养对照组增加了20倍(P < 0.01),从而确定了总细胞因子平衡的差异,并可能影响妊娠预后。结论:多囊卵巢综合征(PCOS)患者和健康人积云细胞培养中细胞因子水平的多向变化可能是囊胚形成和胚胎保存的决定因素。在第5天至第7天,当卵母细胞被选择进入体外受精周期时,对CC产生细胞因子和性类固醇的进一步体外研究尤为重要。利用细胞培养技术研究很少被研究的积云细胞的形态功能特性,将有助于我们更深入地研究PCOS中卵泡发生紊乱的机制,从而改善该疾病的生殖预后。
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引用次数: 0
Combined effect of methotrexate and bifidobacteria metabolites on TNFα AND IFNγ production by human peripheral blood mononuclears 甲氨蝶呤和双歧杆菌代谢物对人外周血单核细胞产生TNFα和IFNγ的联合影响
Pub Date : 2023-08-11 DOI: 10.46235/1028-7221-9839-ceo
E. Ivanova, I. N. Chaynikova, A. V. Bekpergenova, Taisiya A. Bondarenko, O. E. Chelpachenko, I. A. Zdvizhkova, N. Perunova, O. Bukharin
Methotrexate (Mtx) is a first-line drug for the treatment of numerous rheumatic and non-rheumatic disorders, including oncological disdiseases. However, therapeutic efficacy of Mtx is limited by severe toxicity to many organs (myelo-, hepato-, nephrotoxicity, mucositis, enteritis, dysbiosis at various human biotopes, etc.). Recently, a number of studies showed that some metabolites of Bifidobacteria and Lactobacilli are able to enhance effect of chemotherapeutic drugs and limit their toxic properties. The aim of the present work was to study the possible potentiating action of Bifidobacteria cell-free supernatants and methotrexate upon secretion of pro-inflammatory TNF and IFN cytokines by human peripheral blood mononuclear cells (PBMCs). The immunoregulatory effects upon production of TNF and IFNg was evaluated in the in vitro model of cultured PBMC supplemented with Bifidobacteria metabolites, methotrexate, or their combination. Analysis of the combined effect of Bifidobacteria metabolites and Mtx on the cytokine production revealed their synergism towards the key pro-inflammatory cytokines (TNF and IFN). We found an increase against the control cultures (with Mtx only), inhibition of the early pro-inflammatory cytokine TNF production. On the contrary, we revealed an increased secretion of IFN which regulates the effector cells. The results obtained with these cytokines suggest the presence of a potentiating effect of Bifidobacteria metabolites upon anti-inflammatory and immunoregulatory properties of methotrexate. Thus, Bifidobacteria metabolites can be considered a promising agent which potentiates the therapeutic action of methotrexate by suppressing TNF secretion and stimulating IFN by immunocompetent cells. Further studies of the combined effects of Mtx and metabolites from the intestinal microbiota upon the cytokine production by effector cells could be recommended, aiming to enhance therapeutic effect of methotrexate and limit its toxic properties using the Bifidobacteria metabolites.
甲氨蝶呤(Mtx)是治疗多种风湿性和非风湿性疾病(包括肿瘤疾病)的一线药物。然而,Mtx对许多器官的严重毒性(骨髓、肝脏、肾毒性、粘膜炎、肠炎、各种人体生物群落的生态失调等)限制了其治疗效果。近年来,多项研究表明,双歧杆菌和乳酸杆菌的某些代谢物能够增强化疗药物的作用,限制其毒性。本研究的目的是研究双歧杆菌无细胞上清液和甲氨蝶呤对人外周血单核细胞(PBMCs)分泌促炎TNF和IFN细胞因子的可能增强作用。在体外培养的PBMC中添加双歧杆菌代谢物、甲氨蝶呤或它们的组合,评估对TNF和IFNg产生的免疫调节作用。分析双歧杆菌代谢物和Mtx对细胞因子产生的联合作用,发现它们对关键的促炎细胞因子(TNF和IFN)具有协同作用。我们发现对照培养(仅使用Mtx)增加了早期促炎细胞因子TNF的产生。相反,我们发现IFN的分泌增加,从而调节效应细胞。用这些细胞因子获得的结果表明双歧杆菌代谢物对甲氨蝶呤的抗炎和免疫调节特性有增强作用。因此,双歧杆菌代谢物可以被认为是一种有希望的药物,它通过抑制免疫活性细胞分泌TNF和刺激IFN来增强甲氨蝶呤的治疗作用。建议进一步研究甲氨蝶呤和肠道微生物代谢物对效应细胞产生细胞因子的联合作用,旨在利用双歧杆菌代谢物增强甲氨蝶呤的治疗效果,限制其毒性。
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引用次数: 0
Quantitative and phenotypic transformation of CD16+IFNα/βR1-CD119+, CD16+IFNα/βR1+CD119- and CD16+IFNα/βR1+CD119+ neutrophil granulocytes subsets in patients with post-COVID syndrome CD16+IFNα/βR1-CD119+、CD16+IFNα/βR1+CD119-和CD16+IFNα/βR1+CD119+中性粒细胞亚群在covid后综合征患者中的定量和表型转化
Pub Date : 2023-08-11 DOI: 10.46235/1028-7221-9994-qap
I. Nesterova, G. Chudilova, M. G. Atazhakhova, V. A. Matushkina, S. Kovaleva, V. N. Chapurina
In patients who underwent COVID-19, various manifestations of post-COVID syndrome (PCS) are noted, causing the development of disorders accompanying severe viral infections, complicated by chronic fatigue syndrome (CFS) and severe cognitive disorders (CD). Studying the molecular mechanisms of these disorders in the system of neutrophilic granulocytes (NG) in patients with PCS associated with IFN production, receptor function of NG, in particular, their subsets expressing IFN/R, IFNR(CD119), is relevant for the search for therapeutic strategies, restoration and enhancement of the innate immune response after COVID-19. Our objective was to clarify the quantitative and phenotypic characteristics of certain subsets of neutrophil granulocytes, i.e., CD16+IFN/R1-CD119+, CD16+IFN/R1+CD119-, CD16+IFN/R1+CD119+, in peripheral blood of patients with post-COVID syndrome. We have examined 39 patients (24-60 years old) with PCS 3 months after COVID-19 (study group 1, SG1). The comparison group (CG) included 30 volunteers examined over the pre-COVID period. Detection of herpesvirus infections (HSV1, EBV, HHV6, CMV) was carried out in scrapings from the tonsils and the posterior wall of the pharynx. To determine the severity of the clinical PCS symptoms, a questionnaire was used to assess its severity using a point scale. The content and phenotype of NG subsets CD16+IFN/R1-CD119+, CD16+IFN/R1+CD119-, CD16+IFN/R1+CD119+ were assessed by means of FC 500 (Beckman Coulter, USA). In all patients of SG1, clinical manifestations of CFS and CD were revealed, at the average severity rates of 16.0 points (14.75-20.25). When detecting herpesvirus infections, 37.2% had only HSV1 infection; 62.8% of patients showed mixed infection (HSV1, EBV, HHV6), which exhibited more pronounced clinical symptoms. We have noted absence of CD16+IFN/R1+CD119+NG subset and phenotype transformation of CD16+IFN/R1-CD119+NG, CD16+IFN/R1+CD119-NG subsets. Increased density expression of CD16, IFN/R1, CD119 receptors was also found (p1-3 0.05) thus suggesting ability to accept the interferon signaling and response. Reduced infectious burden in the post-COVID period and adequate functioning of the immune system, including the neuroimmunoendocrine regulation mechanisms, should contribute to the functional recovery of various organs, systems, thus neutralizing the PCS manifestations. Therefore, usage of recIFN2b in combination with highly active antioxidants may contribute to development of protective immunity, prevention of acute respiratory viral infections, exacerbation of chronic infections, and restoration of the NG phenotypes followed by restoration of anti-infectious immune balance.
在感染COVID-19的患者中,注意到各种COVID-19后综合征(PCS)的表现,导致伴随严重病毒感染的疾病发展,并发慢性疲劳综合征(CFS)和严重认知障碍(CD)。研究PCS患者中性粒细胞(NG)系统中与IFN产生、NG受体功能相关的这些疾病的分子机制,特别是其表达IFN/R、IFNR(CD119)的亚群,对于寻找治疗策略、恢复和增强COVID-19后的先天免疫反应具有重要意义。我们的目的是澄清某些中性粒细胞亚群的数量和表型特征,即CD16+IFN/R1-CD119+, CD16+IFN/R1+CD119-, CD16+IFN/R1+CD119+,后冠综合征患者外周血。我们检查了39例(24-60岁)在COVID-19后3个月出现PCS的患者(研究组1,SG1)。对照组(CG)包括在covid前期间接受检查的30名志愿者。在扁桃体和咽后壁刮屑中检测疱疹病毒感染(HSV1、EBV、HHV6、CMV)。为了确定临床PCS症状的严重程度,采用问卷调查,采用点量表评估其严重程度。采用FC 500 (Beckman Coulter, USA)检测NG亚群CD16+IFN/R1-CD119+、CD16+IFN/R1+CD119-、CD16+IFN/R1+CD119+的含量和表型。所有SG1患者均有CFS和CD的临床表现,平均严重程度为16.0分(14.75 ~ 20.25)。在检测疱疹病毒感染时,仅有HSV1感染的占37.2%;62.8%的患者表现为混合感染(HSV1、EBV、HHV6),临床症状更为明显。我们注意到CD16+IFN/R1+CD119+NG亚群的缺失和CD16+IFN/R1-CD119+NG、CD16+IFN/R1+CD119-NG亚群的表型转化。CD16、IFN/R1、CD119受体的密度表达也有所增加(p1-3 0.05),提示具有接受干扰素信号和应答的能力。新冠肺炎后感染负担减轻,包括神经免疫内分泌调节机制在内的免疫系统功能正常,有助于各器官、系统功能恢复,从而中和PCS的表现。因此,与高活性抗氧化剂联合使用recIFN2b可能有助于保护性免疫的发展,预防急性呼吸道病毒感染,加重慢性感染,恢复NG表型,随后恢复抗感染免疫平衡。
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Russian Journal of Immunology
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