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摘要： Purpose: T-cell stemness is important for antitumor immunity. The presence in tumor-draining lymph nodes and tumor of stem-like memory T (TSCM) cells and T cells expressing the transcription factor T-cell factor 1 (TCF1), critical in T-cell self-renewal, is associated with improved response to immune checkpoint *** Design: We studied the effects of the tumor-targeting immunocytokine PDS01ADC (NHS-IL12) on peripheral T-cell stemness in murine hosts and in patients with advanced solid tumors. TSCM and expression of the murine T-cell stemness markers stem cell antigen 1 (Ly6a) and Tcf7 were evaluated in na & iuml;ve and tumor-bearing mice receiving murine PDS01ADC (NHS-muIL12). Peripheral blood from patients treated in a clinical trial with PDS01ADC was analyzed for TSCM and expression of TCF1 on T-cell ***: NHS-muIL12 treatment in na & iuml;ve mice increased stem cell antigen 1-positive peripheral T cells with stem-like phenotypes and promoted tumor infiltration of CD8+ T cells displaying increased stemness. In patients with advanced solid tumors, PDS01ADC treatment increased peripheral CD8+ and CD4+ TSCM frequencies and effector memory T cells expressing TCF1, with greater increases associated with disease control. Most peripheral TSCM cells were negative for PD-1 and TIGIT throughout treatment, suggesting their quiescence and self-renewal capacity. Expanded TCF1-negative effector memory CD8+ T cells expressed PD-1 with increased intensity of granzyme B, indicating an activated, cytotoxic ***: PDS01ADC treatment in patients with advanced solid tumors boosts peripheral T cells with stem-like characteristics, correlating with disease stabilization. Further studies combining PDS01ADC with other immunotherapies to synergize with this peripheral burst of T-cell stemness are warranted.

关键词： HPV16 E6突变抗原肽   DC细胞   小鼠宫颈癌模型   治疗效果  

摘要： 目的:建立宫颈癌小鼠模型,探讨负载HPV16 E6抗原肽的树突状细胞(DC)的肿瘤抑制效果及免疫应答反应。方法:选取6~8周龄的健康C57BL/6雌性小鼠,通过皮下注射TC-1细胞的方法建立宫颈癌模型,待肿瘤长至15mm时,将小鼠平均分成3组:control组、DC组、DC+Peptide组。比较各组肿瘤面积,绘制肿瘤生长曲线。从小鼠的脾脏分离出淋巴细胞,流式细胞术检测各组Treg(CD4+CD25+Foxp3+)、IFN-γ表达情况。结果:DC+Peptide组肿瘤面积及重量均显著低于control组及DC组。小鼠脾组织中DC+Peptide组的CD4+IFN-γ、CD8+IFN-γ表达量显著高于control组、DC治疗组,差异有统计学意义(P<0.05);DC+Peptide组Treg数量显著低于control组及DC治疗组,差异有统计学意义(P<0.05)。结论:负载HPV16 E6突变抗原肽的DC细胞,能有效激活小鼠T淋巴细胞,引发特异性抗肿瘤免疫反应,从而抑制肿瘤生长。

关键词： Adalimumab   Cytokine   Etanercept   Infliximab   Paradoxical  

摘要： The inhibition of tumor necrosis factor-α (TNF-α) signaling with inhibitors (TNFi) like infliximab, adalimumab, or etanercept has become a staple of dermatological and rheumatological therapy. Intriguingly, a sizeable minority of patients under TNFi therapy develop alopecia areata (AA), the most common inflammatory hair loss disease, at a higher prevalence than the general population. This appears paradoxical, since the pro-inflammatory Th1 cytokine TNF-α, inhibits hair follicle (HF) growth, but encourages one to reconsider the as yet under-investigated roles of TNF-α in human HF physiology. Because AA only affects HFs whose immune privilege (IP) has collapsed, the occurrence of AA under TNFi t

关键词： Cell lines   CRISPR-Cas9   Haplotype-resolved assembly   MHC   Targeted sequencing  

摘要： The major histocompatibility complex(MHC)region plays a crucial role in immune function and is implicated in various diseases and cancer ***,its high polymorphism poses challenges for accurate genetic profiling using conventional reference ***,we present high-quality,haplotype-resolved assemblies of the MHC region in five widely used tumor cell lines:A549,HeLa,HepG2,K562,and *** oncological studies extensively employ these cell lines,ranging from basic molecular research to drug discovery and personalized medicine *** integrating CRISPR-based targeted enrichment with 10×Genomics linked-read and PacBio HiFi long-read sequencing,we constructed MHC haplotypes for each cell line,providing a valuable resource for the research *** these assembled haplotypes as references,we characterize the aneuploidy of the MHC region in these cell lines,offering insights into the genetic landscape of this critical immunological *** work addresses the urgent need for accurate MHC profiling in these widely used cell line models,enabling more precise interpretation of existing and future genomic and epigenomic *** resource is expected to significantly enhance our understanding of tumor biology,immune responses,and the development of targeted therapies.

关键词： 鸡γ-干扰素   兔源单克隆抗体   流式细胞分选技术   生物膜干涉技术   配对抗体  

摘要： 为提升对禽类细胞免疫反应的检测水平,研究制备高亲和力鸡γ-干扰素(chIFN-γ)兔源单克隆配对抗体。研究利用原核表达的chIFN-γ重组蛋白免疫兔,结合流式单B细胞分选技术筛选获得单克隆抗体,采用生物膜干涉技术和竞争结合试验对抗体的亲和力及配对性能进行系统评估。结果显示:共获得3株单克隆抗体(38-9、56-3和68-7),解离常数(KD)值均达10^(-12)mol/L,显示出极高的亲和力;56-3与38-9和68-7之间无竞争关系,而38-9与68-7之间存在明显竞争,由此可构建56-3/38-9和56-3/68-7两对稳定的配对抗体。研究表明,试验获得的3株单克隆抗体具备高亲和力和良好的配对特性,适用于chIFN-γ的高灵敏免疫检测,为禽类细胞免疫应答监测及chIFN-γ免疫功能研究提供了关键工具。

关键词： 猪繁殖与呼吸综合征病毒   非结构蛋白1α   原核表达   多克隆抗体   生物学特性  

摘要： 【目的】制备非结构蛋白1α(non-structural protein 1α,NSP1α)多克隆抗体,为深入研究NSP1α蛋白功能提供物质基础,并为猪繁殖与呼吸综合征(porcine reproductive and respiratory syndrome,PRRS)防控靶点的选择提供新思路。【方法】以pCAGGS-NSP1α-HA质粒为模板,聚合酶链式反应扩增NSP1α基因,利用相关生物信息学在线软件分析PRRSV SD16毒株NSP1α基因和其他PRRSV毒株NSP1α基因序列同源性、蛋白的理化性质、氨基酸亲-疏水性、跨膜结构域、信号肽序列、磷酸化位点、二级和三级结构。通过EcoR I和Xho I酶切位点将NSP1α克隆至pET-28a-His原核表达载体。测序正确的pET-28a-NSP1α-His质粒转化至大肠杆菌*** BL21(DE3)表达感受态细胞,将诱导表达、纯化复性后的NSP1α蛋白免疫小鼠制备多克隆抗体,Western blot和间接免疫荧光试验(indirect immunofluorescence assay,IFA)鉴定制备的多克隆抗体的反应原性,通过Western blot和激光共聚焦分析NSP1α蛋白亚细胞分布。【结果】信息学分析结果表明,SD16毒株NSP1α蛋白有501个氨基酸,为不稳定的疏水性蛋白,其序列与HLJ毒株序列同源性最高,与I型PRRSV序列同源性最低;NSP1α蛋白无信号肽序列,无跨膜结构域,存在13个磷酸化位点;二级结构主要包含α-螺旋、延伸链和无规则卷曲。将获得的高纯度并且有活性的NSP1α蛋白进行小鼠免疫,检测其血清抗体效价可达1∶64000。Western blot和IFA结果显示,制备的多克隆抗体特异性识别PRRSV NSP1α蛋白。Western blot和激光共聚焦结果显示,NSP1α蛋白主要定位在细胞核内。【结论】成功表达、纯化了PRRSV NSP1α蛋白并制备了鼠源多克隆抗体,该多克隆抗体与PRRSV NSP1α蛋白具有良好的反应活性和特异性。

摘要： Innate lymphoid cells (ILCs) fulfill critical roles in maintenance of tissue-specific homeostasis but have also been implicated in disease pathology when dysregulated. Although they are broadly classified into three core subsets, it is increasingly apparent that ILCs exhibit plasticity in response to microenvironmental factors. Accurate and holistic evaluation of the ILC landscape is critical to understanding the contribution of ILCs to disease pathology. Using high-parameter flow cytometry, we comprehensively interrogated the phenotypic and functional diversity of ILCs in healthy volunteers and patients with severe asthma (SA), assessing the reciprocity between peripheral blood and airway compartments and dissecting the impact of anti-IL-5/5R alpha biologics on these responses. We identified substantial heterogeneity and putative plasticity in human ILC responses, highlighting inherent limitations of conventional enumeration strategies. Deep phenotypic and functional profiling demonstrated a distinct sexual dimorphism in ILC responses in patients with SA. Females displayed an elevated abundance of circulating ILC progenitors, ILC1s, and ILC3s, whereas males presented with diminished ILC2s compared with respective healthy controls. Circulating ILC progenitors inversely correlated with testosterone concentrations. Moreover, we identified a reciprocal influx of all core ILC subsets into the airways of patients with SA, with unbiased multisource clustering identifying a relationship between elevated airway ILC2s and reduced lung function. Last, we showed that anti-IL-5/5R alpha biologics largely ablated airway ILC type 2 cytokine production without affecting core ILC subset abundance in the peripheral blood or airways, identifying a potential mechanism whereby anti-IL-5/5R alpha biologics alleviate clinical disease in patients with SA.

关键词： neutrophil to lymphocyte ratio   platelet to lymphocyte ratio   Takayasu’s arteritis   cardiac compromise   intimate-media thickness   contrast-enhanced ultrasound  

摘要： AIM To assess the utility of NLR,PLR,IMT and contrast-enhanced ultrasound(CEUS)aspredictive markers for monitoring inflammatory responses and the disease activity in cardiac involvementin Takayasu’s *** A cohort retrospective study encompassing 86 patients(43 withcardiac compromise and 43 without)was conducted.A comparative analysis of NLR,PLR,IMT,andCEUS between TA patients with and without cardiac compromise was *** The NLR and PLR of the heart damage group were significantly higher than those of the non heart damagegroup(2.9±1.0 vs.2.1±0.8,P<0.01;166±79 vs.117±51,P<0.01).The IMT and CEUS of the heartdamage group were significantly higher than those of the TA non heart damage group(2.6±0.6 vs.1.5±0.4,P<0.01;2.6±0.5 vs.1.6±0.6,P<0.01).The NLR level of the heart damage group was positivelycorrelated with CRP(r=0.42,P<0.01),and PLR was positively correlated with CRP and CEUS(r=0.34,P<0.05;r=0.35,P<0.05).The results of multiple logistic regression analysis showed that NLR,IMT,andCEUS were independent risk factors for TA and cardiac *** area under the ROC curve for NLRto determine cardiac damage is 0.865,with a cut-off value of 2.265,a sensitivity of 69.8%,and aspecificity of 90.7%.The area under the ROC curve for determining cardiac damage using PLR is 0.812,with a cut-off value of 111.275,a sensitivity of 76.7%,and a specificity of 79.1%.CONCLUSION NLR and PLR,in conjunction with contrast-enhanced ultrasound,can be employed to assessinflammatory response and the disease activity in cardiac involvement in Takayasu’s arteritis.

关键词： 正畸   釉质脱矿   口腔菌群   分泌型免疫球蛋白A  

摘要： 目的 解析正畸患者发生釉质脱矿的关键因素,确定与釉质脱矿发生相关的生物标志物,并探讨青少年正畸患者釉质脱矿与口腔菌群的相关性。方法 选取2组患者,每组各22人,S组佩戴自锁托槽,M组佩戴传统金属托槽。在正畸基线期及3个月后收集龈上菌斑进行菌群分析,同时收集患者唾液并测定分泌型免疫球蛋白A(sIgA)的含量。每月复诊时,对釉质脱矿(EDI)、菌斑指数(PLI)、牙龈指数(GI)、牙周指数(PI)等临床指标进行统计和分析。结果 正畸3个月后,所有患者釉质脱矿发生率显著升高,平均脱矿率较治疗前增加20%以上,其中S组Fusobacterium、Campylobacter、Neisseria和Prevotella菌属的丰度显著增加(P<0.05),而M组Rothia菌属丰度显著上升(P<0.05);Arthrobacter、Bacillus、Acinetobacter、Prevotella、Campylobacter菌属丰度与釉质脱矿评分及菌斑指数评分呈正相关;唾液中sIgA的含量与釉质脱矿的发生呈负相关,S组正畸后唾液sIgA含量显著下降(P<0.01)。结论 佩戴自锁托槽的正畸患者更易发生釉质脱矿,Campylobacter和Prevotella菌属可能是加剧青少年正畸患者釉质脱矿的关键微生物。唾液中sIgA含量的下降可能预示着正畸患者釉质脱矿的风险增加。