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IL-4 and its polymorphism (IL4-589C/T) in cervical neoplasia 宫颈肿瘤组织中IL-4及其多态性(IL4-589C/T
Q4 Medicine Pub Date : 2023-06-01 DOI: 10.15789/1563-0625-iai-2691
T. Abakumova, I. Myagdieva, D. Dolgova, S. Gening, I. Antoneeva, T. Gening
The transition of cervical neoplasia (CIN) to cervical cancer occurs with the active participation of IL-4, for which both pro- and antitumor effects have been shown with tumors of various localizations. The expression of cytokines is regulated at the transcriptional level in the promoter region of the gene. It has been shown that the genotype IL4 (589C/T) (rs2243250) is associated with the development of gastric and breast cancer. The contribution of IL-4 genotypic variations to the development of CIN has not yet been studied. The aim of the study was to assess the risk of developing cervical neoplasia by the presence polymorphism of IL4 (589C/T) and the level of IL-4. The object of the study was circulating neutrophils, serum and genomic DNA of 36 patients with CIN and 20 women without dysplasia (comparison group). Using ELISA, the level of IL-4 was determined in neutrophil lysate and serum. Phagocytic activity and adhesive ability (CD11b) of neutrophils were assessed. Allele-specific real-time PCR using Taq-Man probes was used to analyze of the IL4 589C/T (rs2243250). Statistical processing was carried out using Statistica 13 and Jamovi 1.6.5.0. As a result of the study, it was found that the level of IL-4 in serum and circulating neutrophils in patients with CIN is significantly higher than in the comparison group. The -589C* allele of the IL4 gene and the TT genotype are more common in the group with CIN (55.5%) than in the control (25%). At the same time, a direct relationship was established between the presence of polymorphism and increased adhesive ability and with indicators of the phagocytic number of circulating neutrophils. Analysis of the incidence of IL4 C589T by the «case-control» method showed that the chances of CIN formation in carriers of the -589C allele and the TT genotype were 3.75 (95% CI: 1.013 - 13.880, Chi-square = 4.161, p = 0.042). The -589C* allele and TT IL4 genotype, neutrophil and serum IL-4 levels are associated with HPV infection. Using a binary logistic regression model, we demonstrated the possibility of using IL-4 levels in circulating neutrophils and IL-4 gene polymorphism (589C/T) for the differential diagnosis of patients with CIN (χ2 = 15.6, p = 0.001). Significant significance for their combination was assessed by ROC-curve analysis (IL-4 in neutrophils; IL4 (-589С*), 75% probability. Thus, the IL4 (589C/T) is associated with the adhesive and phagocytic activity of circulating neutrophils. In HPV-infected patients, IL4 gene polymorphism (589C/T) can serve as a marker for early detection and prognosis of CIN.
宫颈瘤变(cervical neoplasia, CIN)向宫颈癌的转变是在IL-4的积极参与下发生的,IL-4在不同部位的肿瘤中均有促瘤和抗瘤作用。细胞因子的表达在基因启动子区域的转录水平上受到调节。已有研究表明,基因型IL4 (589C/T) (rs2243250)与胃癌和乳腺癌的发生有关。IL-4基因型变异对CIN发展的贡献尚未得到研究。该研究的目的是通过IL-4 (589C/T)多态性和IL-4水平的存在来评估发生宫颈瘤变的风险。研究对象为36例CIN患者和20例非发育不良妇女(对照组)的循环中性粒细胞、血清和基因组DNA。采用ELISA法测定血清和中性粒细胞裂解液中IL-4的水平。观察中性粒细胞的吞噬活性和粘附能力(CD11b)。采用Taq-Man探针对IL4 589C/T (rs2243250)进行等位基因特异性实时PCR分析。采用Statistica 13和Jamovi 1.6.5.0进行统计处理。研究结果发现,CIN患者血清及循环中性粒细胞IL-4水平明显高于对照组。il - 4基因的-589C*等位基因和TT基因型在CIN组(55.5%)比对照组(25%)更常见。同时,多态性的存在与黏附能力的增强以及与循环中性粒细胞吞噬数的指标有直接关系。用“病例对照”方法分析IL4 C589T的发病率,结果显示- 589c等位基因携带者和TT基因型携带者形成CIN的几率为3.75 (95% CI: 1.013 ~ 13.880,卡方= 4.161,p = 0.042)。-589C*等位基因和TT IL-4基因型、中性粒细胞和血清IL-4水平与HPV感染有关。使用二元logistic回归模型,我们证明了使用循环中性粒细胞中IL-4水平和IL-4基因多态性(589C/T)对CIN患者鉴别诊断的可能性(χ2 = 15.6, p = 0.001)。通过roc曲线分析(中性粒细胞IL-4;IL4 (-589С*), 75%概率。因此,il - 4 (589C/T)与循环中性粒细胞的粘附和吞噬活性有关。在hpv感染患者中,il - 4基因多态性(589C/T)可作为CIN早期发现和预后的标志。
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引用次数: 0
Hypersegmentation of neutrophil nuclei in peripheral blood of patients with localized and advanced cancer of the larynx and laryngopharynx 局部及晚期喉咽癌患者外周血中性粒细胞核的超分割
Q4 Medicine Pub Date : 2023-06-01 DOI: 10.15789/1563-0625-hon-2715
E. Kologrivova, R. Pleshko, O. Cheremisina, M. Boldyshevskaya
Neutrophilic granulocytes have a wide spectrum of functional activity. In recent years, the functional significance of neutrophils in the development and course of malignant neoplasms has been discussed. It has been shown that neutrophilic granulocytes can play pro- or antitumor activity. The aim of the study was to assess the structural and functional features of neutrophils in patients with varying degrees of prevalence of cancer of the larynx and laryngopharynx. Forty-one patients (aged 35-67) with newly diagnosed cancer of the larynx and laryngopharynx were examined and divided into subgroups according to the TNM classification: the first subgroup (14 patients) with a localized tumor process consisted; and the second subgroup (27 patients) with a widespread tumor process. The relative and absolute number of neutrophils was assessed, and the neutrophil-lymphocyte ratio (NLR) was determined. The content of neutrophils with varying degrees of nuclear segmentation in the blood was calculated, the activity of myeloperoxidase, cationic proteins, alkaline phosphatase, and the degree of neutrophil activation in the NBT test was determined cytochemically. Concentration of interleukin-8 was determined using ELISA. In patients with cancer of the larynx and laryngopharynx the number of neutrophils (p = 0.045) and NLR (p = 0.033), as well as serum concentration of interleukin 8 (p = 0.011), increased compared to healthy individuals. The proportion of cells with hypersegmented nuclei in the neutrophil population (p < 0.001) and cytotoxic potential increased with the spread of tumor process. A direct correlation (r = 0.42, p = 0.03) was found between the T index, which reflects the volume of the tumor, and the content of hypersegmented neutrophils. It can be argued that such a simple and accessible laboratory parameter as the degree of segmentation of the nuclei of neutrophilic granulocytes can be used as one of the criteria to assess and predict the course of the tumor process.
中性粒细胞具有广泛的功能活性。近年来,人们对中性粒细胞在恶性肿瘤发生发展过程中的功能意义进行了探讨。研究表明,嗜中性粒细胞具有促肿瘤或抗肿瘤活性。该研究的目的是评估不同程度喉癌和喉咽癌患者中性粒细胞的结构和功能特征。对41例35 ~ 67岁的新诊断喉癌患者进行检查,并根据TNM分类将其分为亚组:第一亚组(14例)为肿瘤过程局限的亚组;第二亚组(27例)肿瘤进展广泛。测定中性粒细胞的相对数目和绝对数目,测定中性粒细胞与淋巴细胞的比值(NLR)。计算血液中不同核分割程度的中性粒细胞的含量,测定髓过氧化物酶、阳离子蛋白、碱性磷酸酶的活性,测定NBT试验中中性粒细胞的活化程度。ELISA法测定白细胞介素-8的浓度。在喉癌和喉咽癌患者中,中性粒细胞数量(p = 0.045)和NLR (p = 0.033)以及血清白细胞介素8浓度(p = 0.011)比健康人增加。随着肿瘤的扩散,嗜中性粒细胞群中细胞核呈超节段的细胞比例(p < 0.001)和细胞毒性电位增加。反映肿瘤体积的T指数与超节段性中性粒细胞含量呈正相关(r = 0.42, p = 0.03)。可以认为,中性粒细胞细胞核的分割程度这一简单易行的实验室参数可以作为评估和预测肿瘤进程的标准之一。
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引用次数: 0
HDL apoprotein immunization induces T cell-mediated venulitis and inflammation in aorta HDL载脂蛋白免疫诱导T细胞介导的静脉炎和主动脉炎症
Q4 Medicine Pub Date : 2023-06-01 DOI: 10.15789/1563-0625-hai-2699
А. Y. Sidorov, K. Fomina, L. Beduleva
The hypothesis that atherosclerotic processes are mostly caused by immune (autoimmune) mechanisms has recently been gaining attraction. At the same time, the autoimmune hypothesis of atherogenesis has not become generally accepted and requires additional evidence. Previously, we were able to induce changes in the aortic wall similar to those observed in the early stages of human atherosclerosis, and also to produce visceral obesity in normocholesterolaemic Wistar rats by a single immunization with human native high- or low-density lipoproteins. We also found that the immune response to native human HDL causes atherosclerosis-like lesions in the rabbit aorta, such as adipocyte and chondrocyte metaplasia, proteoglycan deposits, and leukocyte infiltration. Atherosclerosis-like lesions developed in the aorta of hnHDL-immunized rabbits against a background of normal blood LDL-cholesterol level. Thus, an immune response against HDL or LDL may be an independent cause of atherogenesis. The aim of this study was to test whether immunization with human HDL apoproteins (apoA1 and apoE proteins) would induce atherosclerosis-like lesions in the aorta of normocholesterolemic Wistar rats. HDL apoproteins were isolated from human or rat plasma. Wistar rats (n = 5) aged 2 months were used for immunization with human HDL apoproteins. HDL apoproteins were administered as a single intradermal injection of 100 mg per rat in incomplete Freund’s adjuvant. Control rats were injected subcutaneously with incomplete Freund’s adjuvant (n = 5). Rats were dissected 25 weeks after immunization. Rat aorta sections were stained with hematoxylin and eosin for light microscopy. T lymphocytes infiltration was determined by immunohistochemical staining with FITC-labeled antibodies specific to rat CD3. CD3+T lymphocytes were detected using an Olympus BX53 fluorescent microscope. The level of antibodies to human and rat HDL apoproteins was determined by indirect enzyme-linked immunosorbent assay. Immunization with HDL apoproteins induced a T cell mediated immune response without production of autoantibodies to HDL apoproteins. The aortic intima and adventitia were infiltrated with T lymphocytes in rats immunized with HDL apoproteins. Pronounced T lymphocytic infiltration was found in all layers of the vein wall in rats immunized with human HDL apoproteins. Thus, immunization with HDL apoproteins causes T cell mediated inflammation of the aorta and venulitis.
动脉粥样硬化过程主要由免疫(自身免疫)机制引起的假说最近受到越来越多的关注。同时,动脉粥样硬化的自身免疫假说尚未被普遍接受,需要更多的证据。以前,我们能够诱导主动脉壁的变化,类似于在人类动脉粥样硬化的早期阶段所观察到的变化,并且通过单次免疫人类天然高或低密度脂蛋白,也可以在正常胆固醇血症的Wistar大鼠中产生内脏肥胖。我们还发现,对天然人HDL的免疫反应会导致兔主动脉出现动脉粥样硬化样病变,如脂肪细胞和软骨细胞化生、蛋白聚糖沉积和白细胞浸润。在血液低密度脂蛋白胆固醇水平正常的背景下,高密度脂蛋白免疫兔的主动脉出现动脉粥样硬化样病变。因此,针对HDL或LDL的免疫应答可能是动脉粥样硬化的独立原因。本研究的目的是测试人类高密度脂蛋白载脂蛋白(apoA1和apoE蛋白)免疫是否会诱导正常胆固醇水平Wistar大鼠主动脉动脉粥样硬化样病变。从人或大鼠血浆中分离出HDL载脂蛋白。用2月龄Wistar大鼠(n = 5)进行人HDL载脂蛋白免疫。每只大鼠在不完全弗氏佐剂中单次皮内注射HDL载脂蛋白100毫克。对照大鼠皮下注射不完全弗氏佐剂(n = 5)。免疫后25周解剖大鼠。用苏木精和伊红对大鼠主动脉切片进行光镜染色。用fitc标记的大鼠CD3特异性抗体免疫组织化学染色检测T淋巴细胞浸润。采用Olympus BX53荧光显微镜检测CD3+T淋巴细胞。采用间接酶联免疫吸附法测定人和大鼠HDL载脂蛋白抗体水平。HDL载脂蛋白免疫诱导T细胞介导的免疫应答,而不产生针对HDL载脂蛋白的自身抗体。高密度脂蛋白载脂蛋白免疫大鼠主动脉内膜和外膜出现T淋巴细胞浸润。人HDL载脂蛋白免疫大鼠静脉壁各层均可见明显的T淋巴细胞浸润。因此,HDL载脂蛋白免疫可引起T细胞介导的主动脉和静脉炎炎症。
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引用次数: 0
Immunomodulatory and neurotropic activities of synthetic peptides in a model of brain injury in rats 合成肽在大鼠脑损伤模型中的免疫调节和嗜神经活性
Q4 Medicine Pub Date : 2023-06-01 DOI: 10.15789/1563-0625-ian-2754
N. B. Serebryanaya, S. N. Shanin, T. A. Filatenkova, E. Fomicheva, A. S. Komlev, O. Shamova
Treatment of consequences of traumatic brain injury (TBI) remains one of the current problems of medicine. To increase the effectiveness of treatment of post-traumatic complications, various drugs are recommended, including the peptide with neuromodulatory activity Semax.The present study aims to determine the presence of neuro- and immunoprotective properties of the synthetic peptide PR5, composed of fragments of proline-rich antimicrobial peptides.The work was performed on male Wistar rats weighing 300-350 g. The “falling weight” model of mechanical brain injury was used, which mainly causes diffuse brain damage. The synthesized peptide PR5, composed of fragments of known proline-rich peptides of animal neutrophils, and the peptide preparation Semax in the form of a 1% aqueous solution were used. The drugs were administered intranasally 1 hour after TBI, then twice a day for 4 days at a dose of 100 mg/kg body weight. Control animals received physiological saline in the same regimen as the peptide preparations.TBI led to a significant decrease in body weight, but in rats receiving the peptide preparation Semax, the decrease in body weight was significantly less than in control animals, and the PR5 preparation completely prevented the decrease in body weight after TBI. After TBI, the proliferative activity of lymphocytes was suppressed and the cytotoxicity of NK cells decreased. In animals treated with peptide preparations, there was no significant suppression of NK cell cytotoxicity, and the proliferative activity of lymphocytes was restored to the level of control animals by day 14 after TBI. Both peptide preparations used contributed to higher locomotor activity, and in animals treated with the PR5 peptide, this type of activity reached the parameters of control animals. The reduction in freezing duration in groups treated with peptide preparations indicates the presence of a sedative effect.The peptide preparation PR5 was active in this series of experiments, showing immunotropic and neuroprotective activity comparable to the Semax preparation. Further studies aimed at confirming the identified types of activity of the peptide preparation PR5 may justify its prospects for clinical use as a new nootropic agent.
创伤性脑损伤(TBI)后遗症的治疗仍然是当前医学的难题之一。为了提高治疗创伤后并发症的有效性,推荐使用多种药物,包括具有神经调节活性的肽Semax。本研究旨在确定由富含脯氨酸的抗菌肽片段组成的合成肽PR5的神经和免疫保护特性的存在。实验对象为体重300-350 g的雄性Wistar大鼠。采用机械性脑损伤的“落重”模型,主要造成弥漫性脑损伤。采用已知动物中性粒细胞富含脯氨酸肽片段合成的肽PR5和1%水溶液形式的肽制剂Semax。这些药物在脑外伤后1小时鼻内给药,然后以100 mg/kg体重的剂量每天两次,连续4天。对照动物以与肽制剂相同的方案给予生理盐水。脑外伤后体重明显下降,但接受肽制剂Semax的大鼠体重下降幅度明显小于对照组,PR5制剂完全阻止了脑外伤后体重下降。脑外伤后淋巴细胞增殖活性受到抑制,NK细胞的细胞毒性下降。经肽制剂处理后,NK细胞毒性无明显抑制,淋巴细胞增殖活性在TBI后第14天恢复到对照动物水平。使用的两种肽制剂都有助于提高运动活性,并且在PR5肽治疗的动物中,这种类型的活动达到了对照动物的参数。在用肽制剂治疗的组中,冷冻时间的减少表明存在镇静作用。肽制剂PR5在本系列实验中表现出活性,表现出与Semax制剂相当的免疫和神经保护活性。进一步的研究旨在确认肽制剂PR5的活性类型,可能证明其作为一种新的促智药的临床应用前景。
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引用次数: 0
MDC/CCL22 depletion in COVID-19 and post-COVID COVID-19和COVID-19后MDC/CCL22消耗
Q4 Medicine Pub Date : 2023-06-01 DOI: 10.15789/1563-0625-mcd-2804
Z. Korobova, A. Totolian
In this article, we explore the role of macrophage-derived chemokine (MDC/CCL22) in COVID-19 immunity. The study included plasma samples of 289 patients with PCR-verified COVID-19 from specialized hospitals. The blood samples were collected at admission, approximately 7 days after the start of infection. Genetic testing of the virus was performed in nasopharyngeal swabs to determine the viral strain for each patient. We also included blood plasma of 69 convalescent patients who had recovered from COVID-19 more than a month prior to the study. Additionally, 51 healthy donors were included in the study as controls. The concentrations of MDC/CCL22 and other cytokines and chemokines were measured with multiplex analysis using Luminex MagPix Technology. The results showed that COVID-19 patients had significantly lower MDC levels in their plasma, regardless of the SARS-CoV-2 strain, compared to healthy donors. This finding suggests that MDC/CCL22 depletion may play a role in COVID-19 immunity. Furthermore, convalescent patients still showed decreased concentrations of MDC/CCL22 more than a month after infection, indicating that this depletion may persist even after recovery. We propose two mechanisms that can explain the reasons leading to MDC/CCL22 depletion. The first is binding and inactivation of this chemokine with SARS-CoV-2 peptides, making it not only undetectable for commercial kits, but also less functionally active. Another mechanism is the dysfunction of its effector cells (e.g., DCs and macrophages). Lymphopenia following COVID-19 can potentially be explained by the absence of MDC/CCL22. This may lead to a shift towards hyperactivation in the inflammatory response, potentially explaining the severity of COVID-19. This research sheds light on the importance of MDC/CCL22 in COVID-19 immunity and highlights the need for further investigation into its role in the disease. Understanding the mechanisms behind MDC/CCL22 depletion could provide new insights into the pathogenesis of COVID-19 and inform the development of potential treatments.
在本文中,我们探讨巨噬细胞来源的趋化因子(MDC/CCL22)在COVID-19免疫中的作用。该研究包括289名来自专科医院的经pcr验证的COVID-19患者的血浆样本。入院时采集血样,大约在感染开始后7天。在鼻咽拭子中进行病毒基因检测,以确定每位患者的病毒株。我们还纳入了69名在研究前一个多月从COVID-19中康复的恢复期患者的血浆。此外,51名健康供体被纳入研究作为对照。采用Luminex MagPix Technology多路分析检测MDC/CCL22及其他细胞因子和趋化因子的浓度。结果显示,与健康供者相比,与SARS-CoV-2菌株无关,COVID-19患者血浆中的MDC水平显着降低。这一发现表明,MDC/CCL22耗竭可能在COVID-19免疫中发挥作用。此外,恢复期患者在感染后一个多月仍显示MDC/CCL22浓度下降,表明即使在康复后这种下降也可能持续存在。我们提出了两种机制来解释导致MDC/CCL22耗尽的原因。首先是这种趋化因子与SARS-CoV-2肽的结合和失活,使其不仅无法被商业试剂盒检测到,而且功能活性也较低。另一种机制是其效应细胞(如dc和巨噬细胞)的功能障碍。COVID-19后淋巴细胞减少可能是由于缺乏MDC/CCL22。这可能导致炎症反应过度激活,这可能解释了COVID-19的严重程度。这项研究揭示了MDC/CCL22在COVID-19免疫中的重要性,并强调了进一步研究其在该疾病中的作用的必要性。了解MDC/CCL22耗竭背后的机制可以为COVID-19的发病机制提供新的见解,并为潜在治疗方法的开发提供信息。
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引用次数: 0
Study of the immunoglobulin and oxidized protein content of semen under infertility 不孕症患者精液免疫球蛋白和氧化蛋白含量的研究
Q4 Medicine Pub Date : 2023-06-01 DOI: 10.15789/1563-0625-sot-2850
N. V. Vavilov, A. P. Godovalov
Among the causes of male infertility, enough attention is paid to oxidative stress, which in turn is a pathogenetic link in the inflammatory process. However, there is practically no information on the content of oxidized modified proteins in the semen, which makes it difficult to study the pathogenesis of diseases of the male reproductive system. In part, protein oxidation may be due to the production of reactive oxygen species by microorganisms, both directly and indirectly through the activation of immune system cells. The aim of the research was to study the level of oxidized modified proteins and changes in immunoglobulin concentrations in the semen under bacteriospermia. A study was made of the ejaculate of 48 men who applied to the clinic for infertility in marriage. The comparison group consisted of 32 practically healthy men who had no growth of microorganisms in the ejaculate samples. When conducting bacteriological analysis, the studied samples were diluted 10 times and used the generally accepted method. The concentration of albumin, immunoglobulins A, M, G, E was determined in the spermatic fluid. The oxidative modification of proteins was evaluated in the reaction with 2,4-dinitrophenylhydrazine. The concentration of oxidized proteins was expressed in nmol/mg of the total protein of the studied biological fluid. The biuret method was used to determine the protein concentration. Statistical analysis of the results was performed using descriptive statistics and Student's t-test for paired data. The concentration of protein in the seminal fluid did not differ significantly among the studied groups. The albumin concentration (16.96±1.28 mg/mL) was statistically significantly lower in the absence of microorganism growth than in bacteriospermia. With bacteriospermia, a decrease in the concentration of IgM and IgA and an increase in the level of IgG were noted. The degree of protein oxidation is maximum when enterobacteria are isolated from seminal fluid. Thus, during the studies it was found that, despite the absence of a clinic, with asymptomatic bacteriospermia, the secretion of immunoglobulins G into the semen is observed. The accumulation of oxidized proteins in the seminal fluid in bacteriospermia has been shown.
在男性不育的原因中,氧化应激引起了足够的重视,而氧化应激又是炎症过程中的一个致病环节。然而,关于精液中氧化修饰蛋白含量的信息几乎没有,这给研究男性生殖系统疾病的发病机制带来了困难。在某种程度上,蛋白质氧化可能是由于微生物产生活性氧,直接或间接通过免疫系统细胞的激活。本研究的目的是研究细菌精子症下精子中氧化修饰蛋白的水平和免疫球蛋白浓度的变化。一项研究对48名因婚姻不孕症向诊所申请的男性的射精进行了研究。对照组由32名实际健康的男性组成,他们的射精样本中没有微生物的生长。在进行细菌学分析时,将所研究的样品稀释10倍,采用普遍接受的方法。测定精浆中白蛋白、免疫球蛋白A、M、G、E的浓度。在2,4-二硝基苯肼反应中评价了蛋白质的氧化修饰。氧化蛋白的浓度以所研究生物液总蛋白的nmol/mg表示。采用双缩脲法测定蛋白浓度。结果采用描述性统计和配对数据的学生t检验进行统计分析。精液中蛋白质的浓度在实验组之间没有显著差异。无微生物组白蛋白浓度(16.96±1.28 mg/mL)显著低于菌精组。在细菌精子症中,IgM和IgA浓度降低,IgG水平升高。当从精液中分离出肠杆菌时,蛋白质的氧化程度最大。因此,在研究过程中发现,尽管没有临床,无症状的细菌精子症,免疫球蛋白G分泌到精液中被观察到。细菌精子症患者精液中氧化蛋白的积累已被证实。
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引用次数: 0
Myeloid-derived suppressor cells as biomarkers of the effectiveness of therapy with new biological agents in axial spondyloarthritis 髓源性抑制细胞作为新生物制剂治疗轴性脊柱炎有效性的生物标志物
Q4 Medicine Pub Date : 2023-06-01 DOI: 10.15789/1563-0625-mds-2696
T. Tyrinova, A. Morenkova, A. V. Fedorova, M. Tikhonova, N. Ilina, O. Chumasova, A. Sizikov
Innate immune cells, including myeloid cells — myeloid derived suppressor cells (MDSCs) — are supposed to play an important role in the pathogenesis of axial spondyloarthritis (AxSp). Myeloid derived suppressor cells represent a heterogeneous population of immature cells capable of suppressing innate and adaptive immune responses with the most pronounced suppressor activity against T cells. Biological disease-modifying antirheumatic drugs (bDMARDs) can reduce the clinical and laboratory disease activity, but their effectiveness varies widely in different patients with AxSp. The present study is aimed at studying MDSCs subpopulations and their suppressive function depending on the response to bDMARD therapy in AxSp. The study included AxSp patients with a disease duration of 16.5 years (median); HLA-B27 (+) status was detected in 79% of cases. All patients received bDMARDs at least the past 12 weeks, including TNF inhibitors (etanercept, certolizumab pegol, adalimumab, or golimumab) or IL-17 inhibitors (secukinumab, ixekizumab, or netakimab). Percentage of granulocytic MDSCs (G-MDSCs, Lin-HLA-DR-CD33+CD66b+), monocytic MDSCs (M-MDSCs, HLA-DRlow/-CD14+), MDSCs of early stage differentiation (E-MDSCs, Lin-HLA-DR- CD33+CD66b-), as well as intracellular expression of arginase-1 was assessed by flow cytometry. Frequency of circulating MDSC subpopulations of patients with a stable response to bDMARDs (responders) did not differ significantly compared to healthy donors. Patients not responding to bDMARDs therapy showed increased relative and absolute number of E-MDSCs compared to healthy donors (pU = 0.01 and pU = 0.02, respectively) and the responders (pU = 0.03 and pU = 0.07, respectively). Increased percentage of E-MDSCs was positively correlated to disease activity — ESR (Rs = 0.821; p = 0.023), CRP (Rs = 0.714; p = 0.07) and ASDASCRP (Rs = 0.829; p = 0.042) in the non-responder group. Responder patients exhibited no correlation between disease activity and circulating MDSCs. The suppressor potential of MDSCs was analyzed by the intracellular expression of arginase-1 molecule which is involved in the inhibition of T cell response. Patients with the stable response were characterized by increased expression of arginase-1 in E-MDSCs compared to donors (pU = 0.02). Non-responders did not demonstrate significant changes in Arg-1 expression, however, the percentage of arginase-1-expressing G-MDSCs was positively correlated to indexes ASDASESR (Rs = 0.857; p = 0.014) and BASDAI (Rs = 0.785; p = 0.036). Thus, E-MDSCs as well as arginase-1 expression in MDSCs may serve as biomarkers of effectiveness bDMARD therapy, and act as potential candidate predictors of response to therapy in AxSp.
先天免疫细胞,包括髓细胞-髓源性抑制细胞(MDSCs),被认为在轴性脊柱性关节炎(AxSp)的发病机制中起重要作用。髓系来源的抑制细胞代表了一种异质的未成熟细胞群,能够抑制先天和适应性免疫反应,对T细胞具有最明显的抑制活性。生物疾病调节抗风湿药物(bDMARDs)可以降低临床和实验室疾病活动,但其效果在不同的AxSp患者中差异很大。本研究旨在研究MDSCs亚群及其在AxSp中对bDMARD治疗反应的抑制功能。该研究纳入病程为16.5年(中位数)的AxSp患者;在79%的病例中检测到HLA-B27(+)状态。所有患者至少在过去12周内接受了bdmard治疗,包括TNF抑制剂(依那西普、certolizumab pegol、阿达木单抗或golimumab)或IL-17抑制剂(secukinumab、ixekizumab或netakimab)。流式细胞术检测粒细胞MDSCs (G-MDSCs, Lin-HLA-DR-CD33+CD66b+)、单核细胞MDSCs (M-MDSCs, HLA-DRlow/- cd14 +)、早期分化MDSCs (E-MDSCs, Lin-HLA-DR-CD33+CD66b -)的百分比以及细胞内精氨酸酶-1的表达。与健康供者相比,对bdmard有稳定反应的患者(应答者)循环MDSC亚群的频率没有显著差异。与健康供体(pU = 0.01和pU = 0.02)和应答者(pU = 0.03和pU = 0.07)相比,bDMARDs治疗无应答的患者E-MDSCs的相对和绝对数量均有所增加。E-MDSCs百分比增加与疾病活动性- ESR呈正相关(Rs = 0.821;p = 0.023), CRP (Rs = 0.714;p = 0.07)和ASDASCRP (Rs = 0.829;P = 0.042)。有反应的患者没有表现出疾病活动性和循环MDSCs之间的相关性。通过细胞内参与抑制T细胞反应的精氨酸酶-1分子的表达分析MDSCs的抑制潜能。与供体相比,反应稳定的患者E-MDSCs中精氨酸酶-1的表达增加(pU = 0.02)。无应答者Arg-1表达无显著变化,但表达精氨酸酶-1的G-MDSCs百分比与ASDASESR指标呈正相关(Rs = 0.857;p = 0.014)和BASDAI (Rs = 0.785;P = 0.036)。因此,E-MDSCs以及MDSCs中的精氨酸酶-1表达可能作为bDMARD治疗有效性的生物标志物,并作为AxSp治疗反应的潜在候选预测因子。
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引用次数: 0
PD-L1 and PD-L2 gene expression in human glioblastoma cells resistant to chemo- and radiotherapy PD-L1和PD-L2基因在化疗和放疗耐药的人胶质母细胞瘤细胞中的表达
Q4 Medicine Pub Date : 2023-06-01 DOI: 10.15789/1563-0625-pla-2693
A. A. Pinevich, N. L. Vartanyan, L. Kiseleva, I. I. Bode, I. Y. Krutetskaya, A. V. Kartashev, V. E. Makarov, T. E. Poneza, I. Smirnov, M. P. Samoilovich
Membrane molecules PD-L1 and PD-L2, ligands of T lymphocytes PD1 receptor, perform immunoregulatory functions. Their binding to the receptor leads to inhibition of proliferation, reduction of cytokine production, cytotoxic response, and apoptosis of T lymphocytes. The cells of many tumors, regardless of their histogenesis, express PD-L1 molecules, thus limiting the development of an anti-tumor immune response. Glioblastomas are highly malignant recurrent tumors of the central nervous system. The main sources of glioblastoma recurrence are resistant tumor cells initially present in gliomas with heterogeneous cellular composition, as well as resistant cells that are formed during therapy. Increasing the dose of cytostatic drugs or radiation during relapse therapy is not effective in glioblastomas. It has been shown for a number of tumors, including ovarian cancer and non-small cell lung cancer, that drugs preventing PD-L1/PD1 interaction are effective in the treatment of neoplasms resistant to chemo- and radiotherapy. Immunotherapy using drugs that inhibit the binding of PD-L molecules to their receptor is considered as a way to overcome the resistance of glioblastomas to therapy. The aim of this work was to assess the level of PD-L1 and PD-L2 gene expression in resistant glioblastoma cells lines A172, R1, T2 and T98G, which resumed proliferation after exposure to the maximum for each line, sublethal doses of cytostatic drugs (fotemustine and temozolomide), fractionated or single gamma irradiation. A172 line belongs to glioblastomas that are highly sensitive to these influences, T98G is a highly resistant cell line, while R1 and T2 lines occupy an intermediate position. In intact glioblastoma A172, R1, and T2 cells the level of PD-L1 and PD-L2 gene expression was equally high, while in T98G cells it was significantly lower. Exposure of A172 and R1 glioblastoma lines to cytostatic drugs or irradiation did not significantly change the level of PD-L1 and PD-L2 genes expression typical for intact cells. In T2 glioblastoma cells, and especially in T98G cells, a significant increase in expression of these genes was found, most pronounced for PD-L2 gene. This increase in expression may indicate an enhanced malignancy of resistant T2 and T98G cells. High expression of the genes responsible for the production of PD-L1 and PD-L2, which limit the cytotoxic response against tumor cells, is a prerequisite for the use of drugs targeted against PD-L1 and PD-L2 for the elimination of resistant cells in glioblastoma.
膜分子PD-L1和PD-L2是T淋巴细胞PD1受体的配体,具有免疫调节功能。它们与受体结合导致T淋巴细胞增殖抑制、细胞因子产生减少、细胞毒性反应和凋亡。许多肿瘤细胞,无论其组织成因如何,都表达PD-L1分子,从而限制了抗肿瘤免疫反应的发展。胶质母细胞瘤是中枢神经系统高度恶性的复发性肿瘤。胶质母细胞瘤复发的主要来源是最初存在于具有异质细胞组成的胶质瘤中的耐药肿瘤细胞,以及在治疗过程中形成的耐药细胞。在复发治疗期间增加细胞抑制药物或放疗的剂量对胶质母细胞瘤无效。研究表明,对于包括卵巢癌和非小细胞肺癌在内的许多肿瘤,预防PD-L1/PD1相互作用的药物对化疗和放疗耐药的肿瘤治疗有效。使用抑制PD-L分子与其受体结合的药物进行免疫治疗被认为是克服胶质母细胞瘤对治疗的耐药性的一种方法。这项工作的目的是评估耐药胶质母细胞瘤细胞系A172、R1、T2和T98G中PD-L1和PD-L2基因表达水平,这些细胞系在暴露于每个细胞系的最大剂量、亚致死剂量的细胞抑制剂(fotemumstine和替莫唑胺)、分次或单次γ照射后恢复增殖。A172系属于对这些影响高度敏感的胶质母细胞瘤,T98G是高抗性细胞系,而R1和T2系处于中间位置。在完整的胶质母细胞瘤A172、R1和T2细胞中,PD-L1和PD-L2基因的表达水平相同,而在T98G细胞中,PD-L1和PD-L2基因的表达水平明显较低。将A172和R1胶质母细胞瘤细胞系暴露于细胞抑制药物或照射下,未显著改变完整细胞典型的PD-L1和PD-L2基因表达水平。在T2胶质母细胞瘤细胞中,特别是在T98G细胞中,发现这些基因的表达显著增加,其中以PD-L2基因的表达最为明显。这种表达的增加可能表明耐药T2和T98G细胞的恶性肿瘤增强。负责PD-L1和PD-L2产生的基因的高表达限制了对肿瘤细胞的细胞毒性反应,这是使用靶向PD-L1和PD-L2的药物消除胶质母细胞瘤中耐药细胞的先决条件。
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引用次数: 0
Attempt to assess direct interactions between tumor burden, myeloid-derived suppressor cells and PD-1- and TIM-3-expressing T cells in multiple myeloma patients 试图评估多发性骨髓瘤患者肿瘤负荷、髓源性抑制细胞和表达PD-1和tim -3的T细胞之间的直接相互作用
Q4 Medicine Pub Date : 2023-06-01 DOI: 10.15789/1563-0625-ato-2760
E. Batorov, T. Aristova, N. Pronkina, V. Denisova, S. Sizikova, G. Ushakova
The avoidance of immune surveillance by malignant plasma cells (PCs) in multiple myeloma (MM) is mediated by different mechanisms, among which an induction of T cell exhaustion and expansion of myeloid-derived suppressor cells (MDSCs) appear to play substantial roles, but it is still a lack of data on possible MDSC-mediated induction of T cell exhaustion. The aim of the present work was to evaluate possible relationship between frequencies of MM PCs, MDSCs and phenotypically exhausted PD-1+ and TIM-3+ T cells in bone marrow (BM) samples and peripheral blood (PB) of MM patients at various disease stages. Peripheral blood (n = 88) and BM samples (n = 56) were obtained from MM patients (newly diagnosed (n = 6), patients in remission (n = 71) and with progressive disease (n = 11)). Frequencies of T cells expressing checkpoint receptors PD-1 and TIM-3, polymorphonuclear MDSCs (PMN-MDSCs, Lin-CD14-HLA-DR- CD33+CD15+/CD66b+), monocyte MDSCs (M-MDSCs, CD14+HLA-DRlow/-), early MDSCs (E-MDSCs, Lin-HLA-DR-CD33+CD15-/CD66b-), and MM PCs (CD45dimCD38+CD138+CD56+CD19-CD117+CD27- CD81-) were assessed with flow cytometry. Circulating and BM-resident PD-1+/TIM-3+T cell subsets, BM E-MDSCs, as soon as MM PCs and serum beta2-microglobulin (B2-M) levels were gradually increased in patients at different stages. Despite that, there were no associations between the markers of tumor load and the studied cell subsets. In patients in remission, BM PMN-MDSCs negatively correlated with CD4+T cells, CD4+PD-1+ and CD8+TIM-3+T cell subsets; there were positive correlations between BM E-MDSCs and CD4+PD-1+TIM-3+ cells and PB M-MDSCs and CD8+PD-1+ and (as a trend) CD8+TIM-3+T cells. We found no associations for the samples of patients at diagnosis and with progression. We can conclude that a possible mutual influence of malignant PCs, MDSCs and PD-1+/TIM-3+T cells is nonlinear, especially during a manifest tumor growth at diagnosis and progression. The detected negative correlations between resident PMN- MDSCs and T cell subsets might be associated with MDSC suppressive function, affecting both predominantly activated PD-1+ cells and exhausted TIM-3+ subsets. The positive correlations between BM E-MDSCs and CD4+PD-1+TIM-3+ cell subset and circulating M-MDSCs and PD-1+ and TIM-3+ CD8+T cells might confirm an ability of MDSCs to induce T cell exhaustion.
多发性骨髓瘤(MM)中恶性浆细胞(PCs)对免疫监视的回避是由不同的机制介导的,其中诱导T细胞耗竭和骨髓源性抑制细胞(MDSCs)的扩增似乎发挥了重要作用,但MDSCs介导的T细胞耗竭可能的诱导仍然缺乏数据。本研究的目的是评估不同疾病阶段MM患者骨髓(BM)样本和外周血(PB)中MM PCs、MDSCs和表型耗竭PD-1+和TIM-3+ T细胞频率之间的可能关系。从MM患者(新诊断患者(n = 6)、缓解患者(n = 71)和疾病进展患者(n = 11)中获取外周血(n = 88)和BM样本(n = 56)。用流式细胞术评估表达检查点受体PD-1和TIM-3的T细胞、多形核MDSCs (PMN-MDSCs、Lin-CD14-HLA-DR- CD33+CD15+/CD66b+)、单核细胞MDSCs (M-MDSCs、CD14+HLA-DRlow/-)、早期MDSCs (E-MDSCs、Lin-HLA-DR-CD33+CD15-/CD66b-)和MM PCs (CD45dimCD38+CD138+CD56+CD19-CD117+CD27- CD81-)的频率。不同阶段患者外周血和脑内PD-1+/TIM-3+T细胞亚群、BM E-MDSCs、MM pc和血清β -微球蛋白(B2-M)水平均逐渐升高。尽管如此,肿瘤负荷标记物与所研究的细胞亚群之间没有关联。在缓解期患者中,BM PMN-MDSCs与CD4+T细胞、CD4+PD-1+和CD8+TIM-3+T细胞亚群呈负相关;BM E-MDSCs与CD4+PD-1+TIM-3+细胞、PB M-MDSCs与CD8+PD-1+和CD8+TIM-3+T细胞呈正相关(趋势)。我们发现在诊断和进展时患者的样本没有关联。我们可以得出结论,恶性PCs, MDSCs和PD-1+/TIM-3+T细胞可能的相互影响是非线性的,特别是在诊断和进展时明显的肿瘤生长期间。驻留PMN- MDSCs与T细胞亚群之间检测到的负相关可能与MDSC抑制功能有关,影响主要活化的PD-1+细胞和耗尽的TIM-3+亚群。BM E-MDSCs与CD4+PD-1+TIM-3+细胞亚群和循环M-MDSCs、PD-1+和TIM-3+ CD8+T细胞之间的正相关可能证实了MDSCs诱导T细胞衰竭的能力。
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引用次数: 0
Clinical and immunological characteristics of medical students depending on the duration and program of study 医学生的临床和免疫学特征与学习时间和课程的关系
Q4 Medicine Pub Date : 2023-06-01 DOI: 10.15789/1563-0625-cai-2753
S. A. Chebotov, R. T. Urazmambetov, I. Andreeva, M. Kharitonova
Chronic psycho-emotional stress can cause dysfunction of neuroimmunoendocrine dysregulation with consequences in the form of a violation of the functional potential of the immune system. Adaptation to new living conditions at the start of studies at a medical university is one of the inevitable circumstances that first-year students overcome. Education under the military training program at a medical university carries an additional stress load in this aspect. Research on the mechanisms of formation of adaptive reactions of the immune system during training under the military training program for officers of the medical service is of undoubted interest. The purpose of the study was to compare the clinical manifestations of immune-mediated pathology and the parameters of adaptive and innate immunity of medical students depending on the length of service and training program. Under observation were 104 medical students, all men, of which 37 were first-year students and 67 were third-year students of a medical university. The subjects of each course were divided into two subgroups depending on the training program. The group of first-year students consisted of 18 people from the military training center (VTC) and 19 people from the medical and preventive faculty (LPF). Among the third-year students of the VUC – 31, LPF – 36. For the clinical characterization of the incidence during the year of study, registration cards for the analysis of immune-mediated pathology were used, the parameters of the immune system at the end of the spring semester were studied using standard methodological approaches. The data obtained indicate that in the first year students with an additional load in the form of a military training program have a more difficult time adapting to learning in comparison with first-year students of the medical faculty. These differences consist in a more frequent and significant clinical manifestation of infectious pathology and are reflected in the functional potential of cellular parameters of innate immunity. The statement of signs of inhibition of the functional potencies of macrophage cells and natural killers in firstyear students of a military training center is an alarming factor in the possible disruption of the adaptive reserves of the immune response system, which probably suggests the need to develop programs to prevent the negative impact of stress-forming factors. By the third year of study, the students of the military training center have the best clinical and immunological indicators of the functioning of the immune system in comparison with the students of the standard educational program of general practitioners. It is likely that during this period the process of psychological adaptation of military medical students is completed.
慢性心理-情绪应激可引起神经免疫内分泌失调的功能障碍,其后果是对免疫系统功能潜力的侵犯。在医科大学开始学习时,适应新的生活条件是一年级学生不可避免要克服的问题之一。在医科大学的军事训练项目下的教育在这方面有额外的压力负荷。军医军官军事训练过程中免疫系统适应性反应形成机制的研究无疑具有重要意义。本研究的目的是比较医学生的免疫介导病理的临床表现和适应免疫和先天免疫参数与服务年限和培训计划的关系。研究对象是104名医科学生,均为男性,其中37名是医科大学的一年级学生,67名是三年级学生。每门课程的受试者根据培训计划分为两个小组。这组一年级学生包括来自军事训练中心(VTC)的18人和来自医学和预防学院(LPF)的19人。在VUC的三年级学生中有31人,LPF有36人。对于研究期间发病率的临床特征,使用免疫介导病理学分析登记卡,使用标准方法学方法研究春季学期结束时的免疫系统参数。所获得的数据表明,与医学院的一年级学生相比,在军事训练项目中有额外负担的一年级学生更难以适应学习。这些差异存在于更频繁和显著的感染病理临床表现中,并反映在先天免疫细胞参数的功能潜能中。在军事训练中心的一年级学生中,巨噬细胞和自然杀手的功能抑制的迹象表明,免疫反应系统的适应性储备可能受到破坏,这可能表明需要制定计划来防止压力形成因素的负面影响。在第三年的学习中,军训中心的学生在免疫系统功能方面的临床和免疫学指标优于全科医生标准教育项目的学生。在此期间,军医学生的心理适应过程可能已经完成。
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Medical Immunology (Russia)
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