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关键词： 结节性皮肤病病毒   L1R△182-204aa蛋白   大肠杆菌表达   单克隆抗体  

摘要： 结节性皮肤病（LSD）是由结节性皮肤病病毒（LSDV）感染牛而引起一种传染病。LSDV基因组长约151 kb，含有156个开放阅读框（ORF），其中L1R基因所编码的L1R蛋白可以诱导机体产生中和抗体，也可作为诊断抗原用于建立诊断方法。为制备LSDV L1R特异性单克隆抗体，本研究将L1R基因截去编码跨膜区（182-204 aa）片段后克隆至pET-28a原核表达载体，诱导表达并纯化重组L1R△182-204aa蛋白，随后将其免疫BALB/c小鼠，通过细胞融合和亚克隆得到杂交瘤细胞株，将其接种小鼠制备腹水并进行效价测定，通过Western blot和IFA验证其特异性。结果显示，利用大肠杆菌表达系统成功表达并纯化获得大小为26 kDa的重组LSDV L1R△182-204aa蛋白，将其免疫小鼠筛选制备了两株能稳定分泌单克隆抗体的细胞株，命名为8A3和8F4，经Western blot和IFA鉴定，两株单克隆抗体不仅能识别大肠杆菌表达的重组L1R△182-204aa蛋白，而且也能与BHK-21细胞表达的重组L1R△182-204aa蛋白发生特异性反应。本研究不仅为LSDV抗体诊断方法的建立和新型疫苗的研发提供了实验材料，同时为深入探索L1R蛋白的生物学功能奠定了基础。

关键词： IL-27   Foxp3   Oral squamous cell carcinoma  

摘要： Objective This study aimed to investigate the expression and clinical significance of interleukin-27 (IL-27) and forkhead box P3 (Foxp3) in oral squamous cell carcinoma (OSCC) tissues, analyze their co-expression correlations, and explore their potential as prognostic biomarkers and therapeutic targets for OSCC.. Methods Immunohistochemistry (IHC) was used to detect the expression of the two subunits of IL-27, IL-27p28 and Epstein-Barr virus-induced gene 3 (EBI3), and Foxp3 in OSCC tissues and normal tissues. Chi-square test was used to analyze the correlation between the expression of the three in OSCC tissues and the clinical pathological parameters of patients. Results The IHC results showed that the expressions of IL-27p28, EB13 and Foxp3 proteins in OSCC tissues were all higher than those in normal control tissues (P < 0.05) Correlation analysis indicated that IL-27 was associated with the presence or absence of lymph node metastasis in OSCC patients, and Foxp3 was related to the clinical stage of the tumor(P<0.05). Their expressions were not related to the patients' age, gender, primary tumor status, degree of differentiation, lesion location, or postoperative recurrence (P > 0.05) . Conclusion The expression levels of IL-27p28, EB13 and Foxp3 in OSCC tissues are significantly upregulated, indicating that the three play important roles in the occurrence and development of OSCC tumors. Furthermore, its high expression is associated with various clinicopathological characteristics of OSCC patients and can be used as a potential indicator for evaluating the prognosis of OSCC patients.

关键词： EGPA   IL5   eosinophil   mepolizumab   benralizumab  

摘要： Introduction: Eosinophilic Granulomatosis with Polyangiitis (EGPA) is a rare necrotizing vasculitis characterized by eosinophilic inflammation that was traditionally treated with corticosteroids associated with other immunosuppressants. Over the last years different biological therapies targeting IL-5/IL-5 receptor have become available and have been employed to tackle this challenging condition. Aim of the present study is to synthesis the evidence on the clinical presentation of this disease and on the efficacy of the newly available therapeutic strategies. Methods: In June 2024 PubMed, Embase and the Cochrane library were searched for studies reporting on EGPA patients being treated by means of different anti IL-5 or anti eosinophils biological therapies. Risk of bias was assessed through the ROBINS-I and RoB2 tools according to study design. Proportion meta-analysis was employed to synthetize data on EGPA clinical manifestations, while data on treatment outcomes was analyzed descriptively due to the high heterogeneity. Results: The present systematic review included 25 studies on a total of 1131 patients. Asthma was present in 99.2% of the patients, Sinonasal involvement in 87.0% and ANCA positivity in 22.8%. The explored treatments consisted in Benralizumab 30 mg every 4 weeks, Mepolizumab 100 mg or 300 mg every 4 weeks and Reslizumab 3mg/Kg every 4 weeks. All the anti-IL-5/IL-5 receptor molecules proved efficacious in remission control and corticosteroid tapering. Conclusion: The available data strongly suggests integrating anti IL-5/IL-5 receptor therapies into EGPA treatment strategies, to enhance patients' outcomes and reduce the long term side effects of prolonged corticosteroid therapy.

关键词： Cytokines   Genetics   Immunology   Innate immunity   Molecular genetics  

摘要： Loss-of-function (LOF) variants in IL6ST, encoding GP130, can cause hyper-IgE syndrome (HIES). Monoallelic LOF variants in IL6ST lead to HIES when located in the intracellular domain downstream of box 1/2 and upstream of the STAT3 phosphorylation sites and the recycling motif, due to their dominant negative (DN) activity. In this region, 2 previously unreported IL6ST variants, p.K702Sfs7* and p.Y759Wfs26*, were identified in 2 families with autosomal dominant (AD) HIES. Both variants were LOF and exhibited DN effects, leading to the accumulation of mutant GP130 on the cell surface. The p.K702Sfs7* mutation was the most upstream N-terminal mutation linked to HIES caused by heterozygous IL6STvariants. Comprehensive screening of IL6ST mutants revealed that most premature terminations downstream of amino acid F641, at the end of the transmembrane domain, resulted in LOF and DN effects via GP130 accumulation on the cell surface. The absence of the recycling motif (positions 782-787) in surface-expressed LOF GP130 led to its accumulation, contributing to the DN effect. The importance of intracellular truncating IL6STvariants can possibly be predicted based on the location of the premature stop codon. GP130 accumulation on the cell surface is a characteristic and potentially diagnostic finding in patients with HIES with heterozygous IL6STvariants.

关键词： 七氟烷   IL-6/HO-1通路   巨噬细胞   迁移   侵袭  

摘要： 目的 探讨七氟醚(SEV)调控诱导巨噬细胞M2极化对食管癌恶性进展的作用及其可能机制。方法 用佛波醇肉豆蔻酸乙酸酯(100 nmol/L)诱导单核细胞THP-1分化为M0型巨噬细胞(Mφ),实验分为对照组、Mφ组、Mφ+SEV组、Mφ+过表达对照(oe-NC)组、Mφ+血红素加氧酶-1(HO-1)过表达(oe-HO-1)组、Mφ+敲低对照(sh-NC)组、Mφ+HO-1敲低(sh-HO-1)组、Mφ+SEV+sh-HO-1组、Mφ+oe-白介素-6(IL-6)组、Mφ+oe-IL-6+sh-NC组和Mφ+oe-IL-6+sh-HO-1组。利用各组巨噬细胞条件培养基分别培养人食管癌细胞KYSE180。q RT-PCR和Western blot检测IL-6、HO-1和巨噬细胞极化相关基因的m RNA和蛋白表达,Transwell实验检测癌细胞迁移和侵袭。结果 在巨噬细胞中,与Mφ组相比,Mφ+SEV组Arg1、 YM1、 IL-4、 IL-6和HO-1表达升高, i NOS表达降低;与Mφ+oe-NC组相比,Mφ+oe-HO-1组Arg1、YM1和IL-4表达增加,iNOS表达降低;与Mφ+sh-NC组比较,Mφ+sh-HO-1组Arg1、YM1和IL-4表达降低,iNOS表达升高;与Mφ+SEV组相比,Mφ+SEV+sh-HO-1组Arg1、YM1和IL-4表达降低,iNOS表达升高。Starbase网站分析显示,在TCGA数据库的食管癌组织样本中IL-6和HO-1之间存在共表达关系。与Mφ+oe-NC组相比,Mφ+oe-IL-6组中IL-6和HO-1表达升高,与Mφ+oeNC组相比,Mφ+oe-IL-6组中IL-6、HO-1、Arg1、YM1和IL-4表达升高,iNOS表达降低;与Mφ+sh-NC组比较,Mφ+sh-IL-6组中IL-6、HO-1、Arg1、YM1和IL-4表达降低,iNOS表达升高;与Mφ+SEV组比较,Mφ+SEV+sh-IL-6组中IL-6、HO-1、Arg1、YM1和IL-4表达降低,iNOS表达上调(P<0.05)。在KYSE180细胞中,相较于对照组,Mφ组和Mφ+SEV组迁移和侵袭细胞数升高;Mφ+SEV组较Mφ组的迁移和侵袭细胞数更高;与Mφ+oe-NC组相比,Mφ+oe-HO-1组迁移和侵袭细胞数升高;与Mφ+sh-NC组比较,Mφ+shHO-1组迁移和侵袭细胞数降低;与Mφ+SEV组相比,Mφ+SEV+sh-HO-1组迁移和侵袭细胞数降低;与Mφ+oe-NC组相比,Mφ+oe-IL-6组迁移和侵袭细胞数增多;与Mφ+oe-IL-6+sh-NC组比较,Mφ+oe-IL-6+sh-HO-1组迁移和侵袭细胞数减少,差异均有统计学意义(P<0.05)。结论 SEV可能通过激活IL-6/HO-1通路促进巨噬细胞M2极化,加速食管癌细胞的迁移和侵袭。

关键词： activated t cells   allogeneic stem cell transplantation   biomarker   chronic GvHD   flow cytometry   predictive  

摘要： Chronic Graft versus Host Disease (cGvHD) is a major late determinant of transplant related mortality (TRM) after allogeneic stem cell transplantation (HSCT). We investigated the predictive value of peripheral blood activated HLA-DR+CD3+ T cells as a novel biomarker. In total, 107 patients were included in this retrospective analysis. Peripheral blood HLA-DR+CD3+ T cells were measured by flow cytometry three months after HSCT. A HLA-DR+CD3+ T cell count <140 cells/μl at month three after HSCT correlated significantly with an increased TRM. A HLA-DR+CD3+ T cell count <100 cells/μl was associated with a higher rate of cGvHD grade 2-3. Subgroup analyses revealed significant results for TRM and cGvHD grade 2-3 for patients with a reduced intensity conditioning (RIC). In summary, low HLA-DR+CD3+ T cells in the peripheral blood three months after HSCT may represent a novel biomarker to identify patients with an increased risk for TRM and cGvHD grade 2-3.

关键词： TNF alpha   non-alcoholic fatty liver   SIRT1   lipid metabolism   fatty liver   choline deficiency model  

摘要： The production of tumor necrosis factor -alpha (TNF alpha) has been associated with fatty liver disease (i.e, hepatosteatosis) for many years. In fact, cytokine production has been thought of as a consequence of hepatic lipid accumulation which then becomes a critical factor in the development of chronic liver pathologies as well as in the pathogenesis of insulin resistance. The purpose of this study was to test the hypothesis that TNF alpha directly regulated lipid metabolism in liver. Wild type mice and mice lacking the receptor for TNF alpha (TNFR1-/-) were fed control diet or a choline-deficient diet. In addition to pro-inflammatory response, choline-deficient diet increased hepatic lipid accumulation and liver injury, serum triglyceride and insulin levels, as well as increased fasting glucose levels in wildtype mice but to a significantly lesser extent in TNFR1-/- mice. Liver perfusion and metabolic cage studies revealed that TNFR1-/- mice exhibited higher rates of lipid oxidation than wildtype mice. Importantly, TNFR1-/- mice have elevated hepatic expression of metabolic and circadian rhythm regulator SIRT1 in comparison to wild type mice. In isolated hepatocytes, TNF suppressed sirt1 expression while inducing expression of DBC1, a known inhibitor of sirt1 function and expression. These data suggest that TNF alpha and possibly other innate immune factors play a critical role in the development of hepatosteatosis and the onset of metabolic syndrome. This data also suggests an interplay among innate immunity, hepatic metabolism, and circadian rhythm in the pathogenesis of metabolic syndrome.

关键词： Satb1   CD4+ T cell differentiation   TH17 cells   autoimmune disease   EAE   IL-2   colitis   scRNA-seq   single-cell MultiOMICs  

摘要： T helper (TH)17 cells are crucial for host defense in barrier organs, and their altered functionality can disrupt tissue homeostasis, increasing the risk of autoimmune diseases. Thus, it is essential to understand the mechanisms controlling TH17 differentiation to develop strategies influencing their role in diseases. Here, we identified Special AT-rich sequence-binding protein 1 (Satb1) as a pioneering factor for TH17 development. Satb1 is highly expressed in TH17 cells, and loss of Satb1 prevents the differentiation of TH17 cells. Consequently, expression of Satb1 in CD4+ T cells is required for the formation of TH17-driven autoimmune diseases. Mechanistically, Satb1 mediates TH17 development through regulating accessibility of the Il2 gene locus and thereby preventing interleukin (IL)-2 signaling early during TH17 differentiation. Hence, suppression of IL-2 expression by Satb1 during TH17 formation is pivotal, suggesting that Satb1 could serve as a novel therapeutic target for treating autoimmune diseases driven by TH17 cells.

摘要： BACKGROUND:Polymyalgia rheumatica (PMR) is an age-related inflammatory disease with shoulder-hip girdle involvement. Magnetic resonance imaging (MRI) reveals extracapsular/entheseal soft tissue involvements in both PMR and spondyloarthritis (SpA) with sacroiliac joint and peri-entheseal spinal bone marrow oedema (BMO) being characteristic of SpA. Therefore, some shared anatomical topography might be expected to result in similar clinical features. Herein we describe the clinical and imaging features of SpA initially diagnosed as PMR. METHODS:Patients followed at Leeds Teaching Hospitals NHS Trust with a diagnosis of psoriatic arthritis (PsA), or axial SpA (axSpA) were screened to identify those initially diagnosed with PMR from 2002 to 2024. Only those patients who retrospectively fulfilled the 2012 EULAR/ACR classification criteria or the Bird criteria for PMR were included. Clinical data relevant to initial PMR diagnosis, imaging features, follow-up and treatment data were collected, as well as radiographic or MRI features that established the final diagnosis. RESULTS:Thirty-one patients [median age in years (IQR): 62 (58-69); 17 females and 14 males] presenting with typical PMR shoulder/hip girdle pain were subsequently classified as SpA-spectrum disorders. The SpA diagnosis was made in 12 patients within three months of presentation, and in 19 patients during the remaining follow-up period [median (IQR): 3 (1-4) years]. Four of 27 tested patients were HLA-B27 positive. BMO on MRI was detected in the spine and/or sacroiliac joints in 20 of 25 imaged patients (80%) (sacroiliac joint: 17 patients [68%], spine: 15 patients [60%]). Clinical resolution with CRP normalisation occurred in 21 of 31 patients following initial glucocorticoid (GC) therapy, but 7 of these 21 initial responders experienced disease flares or CRP elevations. Therapy-wise, disease-modifying antirheumatic drugs (DMARDs) were used in 21 of 31 cases: 8 received conventional DMARDs, and 11 received