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关键词： Autoimmune disease   HLA genotypes   HLA-peptide binding   Han Chinese   Pathogen adaptation  

摘要： Human leukocyte antigen (HLA) genes play a crucial role in the adaptation of human populations to the dynamic pathogenic environment. Despite their significance, investigating the pathogen-driven evolution of HLAs and the implications for autoimmune diseases presents considerable challenges. Here, we genotyped over twenty HLA genes at 3-field resolution in 8278 individuals from diverse ethnic backgrounds, including 4013 unrelated Han Chinese. We focused on the adaptation of HLAs in the Han Chinese by analyzing their binding affinity for various pathogens, and explored the potential correlations between pathogen adaptation and autoimmune diseases. Our findings reveal that specific HLA alleles like HLA-DRB1*07:01 and HLA-DQB1*06:01 confer strong pathogen adaptability at the sequence level, notably for Corynebacterium diphtheriae and Bordetella pertussis. Additionally, alleles like HLA-C*03:02 demonstrate adaptive selection against pathogens like Mycobacterium tuberculosis and coronavirus at the gene expression level. Simultaneously, the aforementioned HLA alleles are closely related to some autoimmune diseases such as multiple sclerosis (MS). These exploratory discoveries shed light on the intricate coevolutionary relationships between pathogen adaptation and autoimmune diseases in the human population. These efforts led to an HLA database at http://***/HLAtyping, aiding searches for HLA allele frequencies across populations.

关键词： biliary tract cancer   cholangiocarcinoma   primary sclerosing cholangitis   STAT3   baricitinib  

摘要： Background/Objectives: Primary sclerosing cholangitis (PSC) is a rare, incurable liver disease characterized by chronic biliary inflammation and fibrosis. PSC is a significant risk factor for biliary tract cancer (BTC). This study aims to evaluate STAT3 expression in BTC and its prognostic significance as well as explore the potential of organoids derived from PSC and liver tumor patients as an in vitro model for testing novel therapeutic strategies in both PSC and BTC. Methods: Fresh tissue samples obtained from 10 PSC patients through targeted endoscopic retrograde cholangiography (ERC) and biopsy samples from liver tumor patients were used to establish organoid cultures. Organoids were treated with different agents and the therapeutic effect was measured by CellTiterGlo. Treatment with the JAK inhibitor baricitinib was followed by the measurement of cytokine concentrations in the supernatant. Archived formalin-fixed paraffin-embedded (FFPE) samples from 55 surgically resected BTC tumors were analyzed for STAT3 expression using immunohistochemistry. Results: We successfully established organoid cultures from all ERC samples. STAT3 protein expression was detected in 56% of tumor samples and 69% of the immune microenvironment. STAT3 positivity in the immune cell compartment was associated with longer disease-free survival, although the multivariate analysis could not confirm its value as an independent prognostic factor. Chemotherapy testing on liver tumor organoids showed various degrees of decreases in viability after treatment with gemcitabine, cisplatin, and cabozantinib. Baricitinib treatment significantly reduced IL-6 and MCP-1 secretion in cholangiocarcinoma Conclusions: The patient-derived organoid model of PSC and liver tumors is a valuable tool for testing novel and established therapeutic strategies, including JAK inhibitors and chemotherapy regimens. STAT3 expression in the immune microenvironment of BTC may serve as a prognostic marker. Further studies

关键词： IL-2   Preterm   Th2   atopic dermatitis   thymus  

摘要： Understanding the reason for the decrease in atopic dermatitis risk in preterm patients may be crucial for the development of prophylaxis and therapeutic measures. The hypotheses include a change in T-helper1/T-helper2/T-helper17 balance, thymus development, and intestinal colonization. This study was designed to compare these parameters between preterm and term patients.  The investigated population comprised 28 preterm and 19 term-born patients. On the 7 day of life, T-helper1/T-helper2/T-helper17 cytokine concentrations were assessed, thymus ultrasonographic examination was performed, and the stool was examined for the presence of pathogenic bacteria. The last two procedures were repeated at 37 weeks of post-menstrual age in the preterm group. There were no significant differences in the concentrations of interferon-γ, tumour necrosis factor-a (TNFa), interleukin (IL)-2, IL-6, or IL-10 after mitogen stimulation between the preterm and term groups. A negative correlation was found between IL-2 and the week of gestation at birth (r= -0.466, p=0.038) and thymus/weight ratio and week of gestation at birth (r= -0.592, p=0.006). IL-6 was negatively correlated with birth weight in preterm group (r -0.694, p=0.008), whereas IL-10 positively with birth weight in term group (r 0.775, p=0.041). Correlations of other investigated cytokines were statistically insignificant. The levels of IL-2 and interferon-γ after phytohemagglutinin stimulation were greater in the subgroup with pathogenic bacteria in the stool at birth (381.38 (148.7-727.4) vs. 13.23 (7.98-197.8) pg/ml; p=0.049; 17.49 (6.53-30.54) vs. 3.37 (1.03-9.82) pg/ml; p=0.037), whereas no significant differences were found between the levels of IL-6, IL-10 or TNFa. The observed associations may indicate an altered pattern of immunological development in preterm and term children.

关键词： human pregnancy   gamma delta T cells   inhibitory KIRs   HLA-C   degranulation  

摘要： Maternal-fetal tolerance mechanisms are crucial during human pregnancy to prevent the immune rejection of the embryo. A well-known mechanism blocking NK-cell cytotoxicity is the interaction of their inhibitory killer-cell immunoglobulin-like receptors (iKIR) with HLA-C molecules on the target cells. In this study, we aimed to investigate the expression of iKIRs (KIR2DL1 and KIR2DL2/3) on the matched decidual and peripheral gamma delta T cells and the localization of HLA-C ligands throughout human pregnancy. The degranulation of gamma delta T cells of pregnant and non-pregnant women in the presence of trophoblast cells was evaluated as well. Our results showed a higher proportion of iKIR-positive gamma delta T cells at the maternal-fetal interface early in human pregnancy compared to the paired blood of pregnant women and full-term pregnancy decidua. In accordance, HLA-C was intensively expressed by the intermediate cytotrophoblasts and decidua-invading extravillous trophoblasts (EVTs) in early but not late pregnancy. Decidual gamma delta T cells during early pregnancy showed higher spontaneous degranulation compared to their blood pairs, but neither decidual nor peripheral gamma delta T cells increased their degranulation in the presence of Sw71 EVT-like cells. The latter were unable to suppress the higher cytotoxicity of gamma delta T cells, suggesting a complex regulatory landscape beyond NK-like activity inhibition.

关键词： 单克隆抗体   尼帕病毒   乳胶微球   检测方法  

摘要： 建立一种基于NiV-G蛋白特异性单克隆抗体的红色乳胶微球免疫层析检测方法。方法：通过纯化后的NiV-G蛋白免疫BALB/c小鼠，采用杂交瘤融合技术制备抗尼帕病毒单克隆抗体，将筛选出的最佳抗体对制备免疫层析试纸条，并评估其灵敏度、稳定性及特异性等。结果显示，经腹水纯化获得5株单抗（E6、C8、F12、D2、G3），筛选确定mAb E6-C8为最佳配对：基于mAb E6（检测线）与红色乳胶微球标记的mAb C8构建试纸条。该试纸条对NiV假病毒检测灵敏度达0.1 ng/mL，与ZIKV、JEV等4种病原体无交叉反应，稳定性良好。在70份猪血清模拟样本中，与ELISA检测结果符合率达94.3%。结果表明，本研究建立的试纸条兼具高灵敏度、强特异性和快速检测优势，为尼帕病毒现场筛查提供了可靠工具，后续需进一步验证其对不同基因型病毒及多样本类型的适用性。

关键词： Neuroinflammation   IL-21   Lipid   Aging   Microglia  

摘要： Neuroinflammation is a key contributor to the onset and progression of neurodegenerative diseases, driven by factors such as viral infections, autoimmune disorders, and peripheral inflammation. However, the mechanisms linking peripheral inflammation or viral infections to neuroinflammation remain poorly understood, limiting the development of effective therapies. Proinflammatory cytokines are implicated in these processes but their effects on brain cells, including microglia, remain insufficiently characterized. Here, we demonstrate that IL-21, a proinflammatory cytokine elevated in autoimmune disorders, chronic viral infections, and Alzheimer's disease, activates microglia and promotes lipid accumulation within these cells. Young, healthy mice injected with IL-21 to mimic chronic exposure exhibited increased proinflammatory cytokine levels and microglial activation in the brain. Notably, microglia in these mice displayed enhanced lipid accumulation, accompanied by upregulation of lipid uptake receptors such as CD36 and TREM-2. These findings were corroborated using the human microglial cell line HMC-3, where IL-21 exposure similarly induced lipid accumulation and increased expression of CD36 and ApoE. Mechanistic investigations revealed that IL-21 upregulates HIF-1 alpha, a transcription factor critical for lipid metabolism and lipid droplet formation. Additionally, we observed elevated IL-21 levels in the circulation of elderly individuals compared to younger counterparts, with IL-21 increases associated with CMV seropositivity. Aged mouse brains mirrored the microglial lipid accumulation and activation patterns seen in IL-21-injected mice. In summary, we identify a novel IL-21-driven mechanism involving lipid accumulation in microglia that contributes to neuroinflammation.

关键词： 猴痘病毒   gp085蛋白   生物信息学分析   原核表达   多克隆抗体  

摘要： 通过生物信息学分析对MPXV gp085蛋白的理化性质、结构、抗原表位等进行预测分析，表达gp085重组蛋白并纯化，制备和分析家兔源多克隆抗体。以MPXV gp085基因为研究对象，应用在线网站对其进行生物信息学分析。采用原核表达系统表达gp085重组蛋白，纯化后通过SDS-PAGE对纯化效果进行验证。将纯化后的gp085蛋白免疫家兔制备多克隆抗体，通过间接ELISA测定抗体效价，通过Western-blot检测多克隆抗体特异性。生物信息学分析表明，MPXV gp085蛋白为稳定性亲水蛋白，无跨膜区和信号肽，有3个线性表位；在9种正痘病毒中，对应MPXV gp085的同源基因序列一致性高，其中与牛痘病毒的亲缘关系最近。MPXV gp085重组蛋白主要以包涵体形式表达，大小约为17 kDa。间接ELISA检测结果显示其多克隆抗体效价达1:51 200，Western-blot结果表明其多克隆抗体能够特异性识别gp085蛋白。MPXV gp085重组蛋白具有较好的抗原性，免疫家兔后能刺激产生高滴度的抗体，这为进一步研究MPXV gp085蛋白的生物学功能以及快速诊断技术的开发提供生物材料和理论依据。

关键词： 猪伪狂犬病病毒   单克隆抗体   gE蛋白   表位鉴定  

摘要： 为了制备猪伪狂犬病病毒（PRV）g E糖蛋白的单克隆抗体（MAb），本研究将PRV JM株gE蛋白主要抗原表位区段（aa52～aa238）的基因序列构建重组表达质粒pET-32a-gE，经原核表达及纯化重组蛋白，并免疫BALB/c小鼠。取3次免疫小鼠脾脏制备脾细胞，与小鼠骨髓瘤细胞（SP2/0）细胞融合。通过ELISA筛选杂交瘤细胞株，western blot和间接免疫荧光试验（IFA）鉴定MAb的反应性。使用MAb亚类鉴定试剂盒检测重链和轻链类型。以gE截短合成多肽为抗原，采用间接ELISA方法鉴定MAb识别的抗原表位。并利用MAb鉴定PRV野生型病毒及基因缺失病毒PRV JMΔgE/TK株中gE蛋白的表达情况。结果显示，经间接ELISA方法筛选到1株稳定分泌gE蛋白的单克隆细胞株，命名为4C6B。western blot和IFA均表明本研究筛选的MAb能够特异性识别PRV感染的PK-15细胞中表达的gE蛋白；MAb亚类鉴定结果显示重链类型为IgM型，轻链类型为κ链。间接ELISA结果显示MAb能够识别一个线性B细胞表位131ACHPDLVL GRACVPEAPEMG150。利用制备的MAb腹水进行PRV野毒和gE基因缺失毒株的鉴定，结果显示该MAb能够有效鉴别出PRV JM野毒株与g E缺失株。本研究制备的PRV gE蛋白的4C6B MAb可以用于特异性鉴别PRV JM野毒株和gE基因缺失株，同时，也可以为PRV诊断方法的建立、PRV基因缺失疫苗的构建以及gE蛋白的生物学功能研究提供材料。

关键词： 多发性骨髓瘤   免疫治疗   单克隆抗体   嵌合抗原受体T细胞   抗体药物耦联物   双特异性抗体  

摘要： 多发性骨髓瘤（multiple myeloma，MM）是一种以恶性浆细胞增殖为特征的血液系统肿瘤。目前，蛋白酶体抑制剂（proteasome inhibitors，PIs）、免疫调节剂（immunomodulatory drugs，IMiDs）和自体造血干细胞移植（autologous hematopoietic stem cell transplantation，ASCT）是改善MM患者生存预后的主要手段，但随着治疗的深入，大多数患者仍面临难治和复发问题，治愈之路依然充满挑战。近年来，单克隆抗体（monoclonal antibodies，mAbs）、嵌合抗原受体T细胞（chimeric antigen receptor T-cells，CAR-T）、抗体药物耦联物（antibody-drug conjugates，ADCs）和双特异性抗体（bispecific antibodies，BsAbs）均在临床研究和/或临床应用中取得了良好的疗效，这些基于抗体和细胞的免疫治疗为难治复发性MM（refractory and relapsed multiple myeloma，RRMM）患者带来了新的希望。各类免疫治疗产品在激活免疫细胞、靶向肿瘤以及改善患者预后方面各有优势，但同时也面临安全性和耐药性的挑战。未来，随着分子生物学和抗体工程技术的不断进步，新型免疫治疗产品有望通过联合疗法等多种策略实现协同增效，从而为RRMM患者带来更长久的生存获益和生活质量的改善。本文旨在综述多发性骨髓瘤免疫治疗的研究成果与现存挑战，探讨单克隆抗体、CAR-T、抗体药物耦联物和双特异性抗体的作用机制、临床应用现状及未来发展趋势，为相关领域研究者和临床医生提供一个全面和系统的参考，推动MM精准免疫治疗的发展。

摘要： Several multifactorial pathologies in ophthalmology that affect the anterior segment of the eye are partly inflammatory. To better understand the role and impact of inflammation in dry eye and corneal healing, many research teams have used in vitro models to mimic different aspects of these diseases. Several in vitro models have been developed to elucidate the signaling cascades involved in pathogenesis. They also offer the experimental flexibility to adjust environmental parameters, facilitating the validation of innovative therapies and the identification of new pharmacological targets. This review focuses on two-dimensional in vitro models, but also highlights the progress made in 3D models obtained by tissue engineering, which mimic inflammation in these ocular pathologies. The origin of the cells (human or animal), their tissue source, the type of cells (epithelial, endothelial, vascular, conjunctival), as well as the various experimental conditions used to mimic an inflammatory aspect according to the stages of progression of these pathologies, are thoroughly reported in this review of the literature.