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关键词： X射线   辐照   淋巴细胞   血常规   剂量  

摘要： 考察了X射线辐照抑制血液中淋巴细胞增殖的效果,以确定预防输血相关性移植物抗宿主病(TA-GVHD)的最佳辐照剂量。实验在150 kV管电压、8 mA束流下进行,通过调整辐照距离和时间控制剂量。结果显示,40 Gy剂量下,淋巴细胞增殖抑制率达92.62%±1.32%,且对血常规参数无显著不良影响。当剂量提高到70 Gy时,尽管淋巴细胞增殖抑制率没有显著提升,却导致红细胞内钾离子浓度急剧升高,引发溶血。这表明高剂量辐照会损害红细胞完整性。因此,40Gy被认为是全血X射线辐照的最佳剂量,能在有效抑制淋巴细胞增殖的同时保持血液成分稳定性。此剂量下,能够有效降低TA-GVHD风险,且不会对血液质量造成显著影响。该研究为临床输血安全提供了重要的实验数据和指导,有助于优化辐照参数,提高输血安全性及有效性,减少不良反应的发生。

关键词： HLA   T cell receptor   computational protein design   human leukocyte antigen   peptide antigen  

摘要： Recognition of epitopic peptide antigens presented on class I major histocompatibility complex (MHC-I) proteins by T cell receptors (TCRs) forms the cornerstone of immune surveillance, leading to a plethora of adaptive immune responses. Characterization of TCR:peptide/MHC-I interactions is critical for understanding immune recognition, and developing immunotherapies, but the large variation in docking orientations of TCRs on their peptide/MHC-I targets challenges structural modeling. NMR spectroscopy could potentially resolve this ambiguity, but the large size of the TCR:peptide/MHC-I complex limits data quality. Here, we demonstrate that a designed MHC-I protein, SMART A*02:01, enables facile solution mapping of MHC-I:TCR interactions at scale. Our approach can be combined with computational modeling and structure-guided engineering to aid the development of TCR-based therapeutics.

关键词： Asthma   Immunology   Inflammation   Pulmonology  

摘要： The mechanism of neutrophilic and mixed neutrophilic-eosinophilic asthma is poorly understood. We found that extracellular DNA and nucleosomes (Nuc) were elevated in the airways from neutrophilic-eosinophilic asthma patients and correlated with bronchoalveolar lavage neutrophils. Bronchial tissue from neutrophilic-eosinophilic asthma expressed increased DNA sensor-positive cells. Intranasally administered DNA did not induce airway hyperreactivity (AHR) or any pathology but induced AHR and neutrophilic-eosinophilic inflammation when co- administered with the allergen Alternaria (Alt). Nuc alone induced anti-inflammatory/defensive genes whereas the Nuc-Alt combo increased TNF and innate cytokines. The Alt-Nuc phenotype was abolished in Cgas-/-, ALR-/-, Sting-/-, LysMCre:Stingf/f, IL7RCre:Rorαf/f and Tnfr2-/- mice. Alt, unexpectedly, played an essential role in the Nuc-induced phenotype. It abrogated Nuc-induction of anti-inflammatory genes, facilitated Nuc uptake, induced ILC2s, which, in presence of Nuc, produced high levels of TNFα and promoted neutrophilic infiltration. We established a paradigm where allergens inhibit the anti-inflammatory effects of DNA/Nuc and facilitate STING-TNFα-driven neutrophilic-eosinophilic inflammation in asthma.

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, two previously unreported IL6ST variants, p.K702Sfs7* and p.Y759Wfs26*, were identified in two families with autosomal dominant (AD) HIES. Both variants were LOF, 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 IL6ST variants. 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 IL6ST variants 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 HIES patients with heterozygous IL6ST variants.

关键词： 非洲猪瘟病毒   单克隆抗体   M1249L蛋白  

摘要： 本研究旨在制备非洲猪瘟病毒（ASFV）M1249L蛋白的单克隆抗体并分析其亚型、灵敏性及反应性。首先对M1249L（673 aa～1249 aa）序列进行密码子优化，并将该片段插入pET-28a载体中，构建原核表达质粒；经过转化、挑取单克隆、诱导表达及蛋白纯化，用纯化后的M1249L截短蛋白免疫小鼠；经细胞融合、有限稀释法筛选分泌抗体的杂交瘤细胞株，进行亚型鉴定和腹水制备。采用蛋白质免疫印迹（Western-blot）和间接免疫荧光试验（IFA）检测该单抗的反应性。结果显示，该单抗1D6重链属于IgG1型，轻链为Kappa型，腹水抗体效价可达1∶1 024 000。Western-blot结果显示，该单克隆抗体能够特异性识别ASFV感染后不同时间点猪肺泡巨噬细胞（PAMs）表达的M1249L蛋白以及HEK 293T细胞转染不同质粒量后表达的M1249L蛋白。IFA结果显示，在ASFV感染的PAMs细胞中出现绿色荧光，表明有良好的反应性。综上所述，本研究成功制备出M1249L单克隆抗体，为后续研究M1249L蛋白的生物学功能和ASFV诊断检测奠定了基础。

关键词： 白芍总苷   自身免疫性甲状腺炎   微小核糖核酸-155   细胞因子信号转导抑制蛋白1   T淋巴细胞   免疫调节  

摘要： 目的 探讨白芍总苷通过调控miR-155表达对自身免疫性甲状腺炎（autoimmune thyroiditis,AIT） Th17/Treg平衡的影响。方法 将60只雌性SD大鼠随机分为空白对照组、模型组、白芍总苷高、中、低剂量组及西药对照组（硒酵母组）,造模后给药8周,HE染色观察甲状腺组织病理改变,RT-PCR法检测大鼠甲状腺中miR-155表达量,双荧光素酶法验证miR-155对SOCS1的靶向作用,ELISA法检测外周血清TGAb、TPOAb、IL-10、IL-17水平,Western blot法检测甲状腺SOCS1表达,流式细胞法检测脾脏CD4+细胞中Th17、Treg占比。结果 与空白组对比,模型组大鼠甲状腺miR-155表达明显增加,血清TGAb、TPOAb、IL-17水平明显提高,Th17占比增加;IL-10水平明显降低,Treg占比减少,SOCS1蛋白表达降低,差异有统计学意义（P<0.01）;与模型组比较,白芍总苷高、中、低剂量组及硒酵母组miR-155表达量降低,TGAb、TPOAb、IL-17水平降低、Th17占比减少,IL-10水平增加,Treg占比增加,SOCS1表达增强,差异有统计学意义（P <0.01）,且白芍总苷中剂量组的改善更为明显。结论白芍总苷可下调miR-155表达,调节Th17/Treg平衡,减轻炎症反应,发挥免疫调控的作用。

关键词： bacterial vaginosis   G. vaginalis   IL-33   IgA   TGF-β  

摘要： Introduction:Bacterial vaginosis (BV) is a common gynecological disease characterized by an abnormal increase in vaginal secretions, odor and itching. The pathogenesis of BV is not fully understood, but it is believed that the disruption of the mucosal immune system plays a key role. We investigated the role of IL-33 in preventing BV and explored the mechanism by which IL-33 regulates intravaginal IgA. Methods:Protein levels of IL-33 and IgA, and the pH value of vaginal secretions in healthy donors and patients with BV (14 vs 14) were determined by ELISA. -induced bacterial vaginosis mouse model was established using wild-type (WT) and IL-33 knockout (KO) mice. Protein levels of IL-33, IgA and TGF-β, the pH value of vaginal secretions, and Gram-staining were measured in vivo and in vitro to investigate the role of IL-33 in BV progression. Results:IL-33 and IgA were significantly decreased in vaginal secretions of patients with BV. IL-33 deficiency aggravated BV induced by in a mouse model, while IL-33 supplementation prevented it. IL-33 modulated intravaginal IgA expression through the TGF-β signaling pathway in B cells. Conclusion:IL-33 prevents -induced BV by modulating intravaginal IgA expression through the TGF-β signaling pathway in B cells.

关键词： Hantaan virus (HTNV)   Nucleocapsid protein (NP)   pan-MHC-II epitope   Immunoreactivity  

摘要： During viral infection, CD4+ helper T-cell is indispensable for the establishment of the humoral immune protection, CTL activation, and even long-term memory response. It requires MHC-II molecules in viral structural antigens process and presentation. HTNV NP epitopes exhibited high affinity to both of HLA-II superfamilies and H-2-I genes in the present study. Immunogenicity and conservation analyses identified 34 selective epitopes, later validated by molecular docking (MD) with MHC-II structures. NP 15-mer peptides and MHC-II haplotypes were found to interact bidirectionally through hierarchical clustering. In brief, epitopes that exhibit immunoreactivities for a wide range of MHC-II molecules reflect the biomedical practice of vaccination, while haplotype clusters reflect individual differences in T-cell antigen presentation. Then, 11 HTNV variants showed three amino acid substitutions in three epitopes, with little impact on their pan-HLA-II immunoreactivity. Safety analyses indicated that the 34 selective epitopes exhibit favorable safety profiles for potential applications. Finally, we validated the immunogenicity of the selective epitopes using ELISA, ELISpot, and flow cytometry. In conclusion, our work provides a comprehensive assessment of the pan-MHC-II immunoreactivity of HTNV NP and lays the theoretical and technical foundations for the development of protective epitope vaccines in the context of population immunity.

关键词： Clinical Trial   Rheumatology   THERAPEUTICS  

摘要： INTRODUCTION:Biological disease modifying antirheumatic drugs (bDMARDs) have a central role in the treatment of rheumatoid arthritis (RA). Tumour necrosis factor inhibitors (TNFis) are commonly used as first-line agents, while non-TNFis (tocilizumab, abatacept and rituximab) have shown to be non-inferior to TNFis in head-to-head trials. In case of TNFi inadequate response, using other mechanisms of action provides a better response than using an alternate TNFi. Which non-TNFi bDMARD administered subcutaneously to allow for ambulatory management to choose in case of first line TNFi inadequate response has not been tested in a randomised clinical trial, while observational data support a potential superiority of tocilizumab over abatacept. METHODS AND ANALYSIS:The SUNSTAR (SUbcutaNeouS Tocilizumab vs Abatacept in TNF Alpha inadequate responders for the treatment of Rheumatoid arthritis) study is a 52-week prospective, randomised, multicentre, open-label, superiority phase IV trial comparing subcutaneous tocilizumab with abatacept in a 1:1 ratio. Patients with active RA (Disease Activity Score-erythrocyte sedimentation rate >3.2 and Clinical Disease Activity Index (CDAI) >10) and with inadequate 3-month response to a first or second TNFi are included in 25 centres in France. The primary outcome is the CDAI improvement at week 24. Intention-to-treat analysis will be applied primarily. The secondary outcome is a composite outcome of the percentage of responders defined as a CDAI<10, no use of rescue treatment after week 12 and no experimental treatment change, at weeks 12, 24 and 52. ETHICS AND DISSEMINATION:Ethics approval was obtained from the committee for the protection of persons (Comité de protection des personnes Sud Méditerranée I 17.00608.001744). The findings from this study, whether positive or negative, will be published in peer-reviewed journals and will be presented at national and international conferences. The results will inform future recommendations on

关键词： 支气管哮喘   烧山火针法   电针疗法  

摘要： 目的：探讨烧山火针法治疗支气管哮喘的疗效及对血清白介素-4(IL-4)、γ干扰素(IFN-γ)和白介素-6(IL-6)水平的影响。方法：挑选本院予以相应治疗的支气管哮喘患者，随机分成两组，对照组32例给予常规针刺疗法，观察组28例给予烧山火针法，治疗后比较两组疗效、哮喘生活质量调查问卷评分(AQLQ)和哮喘控制测试表(ACT)评分、肺功能、中医症候积分、血清IL-4、IFN-γ和IL-6水平。结果：观察组总有效率92.86%(26/28),明显高于对照组的75.00%(24/32),差异有统计学意义(P<0.05);AQLQ评分和ACT评分方面，治疗后两组均改善，差异有统计学意义(P<0.05),且观察组改善更明显，差异有统计学意义(P<0.05);第1 s用力呼气量(FEV1)、第1 s用力呼气量与用力肺活量比值(FEV1/FVC)以及峰流速(PEF)值方面比较，两组治疗后均升高，差异有统计学意义(P<0.05),并且观察组升高更明显，差异有统计学意义(P<0.05);中医的主症、次症和总积分方面，两组治疗后均降低，差异有统计学意义(P<0.05),且观察组降低更明显，差异有统计学意义(P<0.05);治疗后两组血清炎性因子水平均明显降低，差异有统计学意义(P<0.05),且观察组降低更明显，差异有统计学意义(P<0.05)。结论：烧山火针法治疗支气管哮喘具有较好的疗效，并且能够缓解炎症反应。