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关键词： Mucosal immunology  

摘要： The airway epithelium plays a crucial role in maintaining lung homeostasis, and its dysregulation is often linked to various lung diseases, including asthma. Ciliated cells, abundantly present in the mammalian airway epithelium, have a critical function in clearing inhaled particles and pathogens. We show here that ciliated cells constitutively express IL-17D, which functions as an immune brake in limiting allergic inflammation in murine models of asthma. Mechanistically, IL-17D functions to prevent influx of classical monocytes into the lung and their subsequent conversion to pathogenic alveolar macrophages, through binding to CD93. Deficiency in Il17d or Cd93 increased the expression of chemokine receptors on classical monocytes, including CCR6, thereby enhancing their recruitment to the lung and type 2 inflammation. Our study thus reveals an unexpected protective role of ciliated cells and IL-17D in lung immune responses and asthma, which can be further explored for treating related diseases.

摘要： PURPOSE:The phase II NIVOREN GETUG-AFU-26 study assessed the safety and activity of nivolumab in a real-world population with pretreated advanced clear cell renal cell carcinoma (aRCC). A comprehensive translational research program was conducted including blood and tissue-based analyses. Herein we assessed circulating factors at baseline, their association with intratumoral immune contexture and outcomes. EXPERIMENTAL DESIGN:Baseline blood samples were prospectively collected from 353 patients included in the NIVOREN trial. A cohort of 80 patients including 40 responders and 40 with primary progressive disease, was used for biomarker discovery, with fourteen soluble factors (VEGF,VCAM-1, IL-6, IL-7, IL-8, IL-10,APRIL, BAFF, 4-1BB, BCA, SDF-1, MDC, IFN-gamma and TNF-alpha) assessed for association with overall survival (OS) (discovery set). Candidate biomarkers were subsequently assessed in the remaining 273 patients for association with outcomes (validation set). Gene expression signatures were assessed on baseline tissue samples using RNA sequencing. RESULTS:Five candidate biomarkers, IL-6,IL-7,IL-8,VEGF, and 4-1BB, were significantly associated with OS in the discovery set. We confirmed in the validation set a significant adverse association between OS and higher baseline levels of IL-6, IL-8, VEGF, with respective hazard ratios of 3.17 (95%CI 2.29-4.40), 2.11 (95%CI 1.60-2.80), and 1.36 (95%CI 1.04-1.79). Higher IL-6 and IL-8 levels were associated with an intratumor IMmotion myeloid gene expression signature (p=0.004 and p=0.041 respectively). CONCLUSIONS:Elevated baseline circulating IL-6/IL-8/VEGF are associated with worse outcomes in nivolumab-treated aRCC. Circulating IL-6 and IL-8 showed strongest association with outcomes and correlated with a myeloid tissue contexture, providing guidance for patient selection in future trials.

关键词： 药物超敏反应综合征   阿司匹林   吲哚布芬   药物不良反应   药源性疾病  

摘要： 本文报告了1例冠心病患者应用阿司匹林和吲哚布芬抗血小板治疗过程中发生交叉药物超敏反应（drug-induced hypersensitivity syndrome，DIHS）的病例，旨在提高临床诊疗团队对DIHS的认识及鉴别能力。通过对2024年12月心内科收治的一名DIHS患者临床资料进行回顾性分析（包括不良反应史、临床表现、病情进展、治疗及转归），明确患者交叉DIHS的发生与环氧酶-1（COX-1）通路相关。结合对同类药物DIHS的文献检索，提示既往发生过阿司匹林相关DIHS的人群，应用吲哚布芬时需谨防交叉过敏的发生。

关键词： 型草鱼呼肠孤病毒   VP4蛋白   单个B细胞技术   单克隆抗体制备  

摘要： 【背景】Ⅱ型草鱼呼肠孤病毒（GCRV-Ⅱ）严重危害中国草鱼（Ctenoparyngodon idellus）养殖业的健康发展。【目的】本研究基于单个B细胞抗体制备技术平台，结合哺乳动物表达系统，旨在制备靶向GCRV-Ⅱ VP4蛋白的基因工程抗体。【方法】通过构建GCRV-Ⅱ VP4基因表达载体，在真核表达系统中制备重组VP4蛋白；将纯化后的重组蛋白作为免疫原多次免疫新西兰白兔，分离出脾脏单个抗原特异性记忆B细胞；随后通过聚合酶链式反应（PCR）扩增抗体可变区、恒定区基因片段，构建重组表达载体并转染HEK293F细胞进行单克隆抗体表达。最终通过酶联免疫吸附试验（ELISA）、蛋白质免疫印迹（Western blot）和免疫荧光（IF）评估抗体的抗原结合特性。【结果】通过对分离培养的单个B细胞上清液筛选，利用ELISA筛选出对VP4蛋白具有较好结合能力的3种B细胞克隆（1D5、1F9、1H1），并结合IF实验验证了其能结合病毒感染组织中的VP4抗原。进一步通过抗体重组表达获得了3株抗VP4重组单克隆抗体（1D5、1F9、1H1），其ELISA效价均达1∶128 000。Western blot分析结果显示，1D5、1F9和1H1单克隆抗体能特异性识别GCRV-YX246感染的脑细胞中65.4 kDa的蛋白质条带，该分子量大小与病毒VP4蛋白理论分子量一致。IF实验结果也进一步证实1D5、1F9和1H1可特异性识别GCRV-YX246毒株感染的草鱼头肾组织切片中的病毒抗原。【结论】本研究成功制备并筛选抗GCRV-Ⅱ VP4重组单克隆抗体，为GCRV-Ⅱ病毒快速诊断试剂盒的研发和VP4蛋白功能研究奠定基础。

关键词： 牛病毒性腹泻病毒   Erns蛋白   单克隆抗体   ELISA  

摘要： 为了制备牛病毒性腹泻病毒（BVDV）单克隆抗体及建立抗原捕获ELISA检测方法，采用大肠杆菌原核表达的BVDV Erns截短蛋白免疫BALB/c小鼠，三免后利用细胞融合和杂交瘤筛选，选用效价高的单克隆抗体，通过条件优化建立BVDV抗原捕获ELISA检测方法，对其特异性、敏感性和重复性进行了验证，并与IDEXX商品化试剂盒及RT-PCR进行临床样品符合。结果显示，获得2株效价较高的IgG1型单克隆抗体杂交瘤细胞2B3、2B10，轻链均为κ链。选用效价更高的2B10单克隆抗体作为检测抗体，兔源BVDV多克隆抗体作为捕获抗体，建立的BVDV抗原捕获ELISA与临床常见病原（牛轮状病毒、牛冠状病毒、牛副流感病毒3型、牛呼吸道合胞体病毒、牛支原体）无交叉反应，最低可检测102.53TCID50的BVDV病毒液，批内和批间变异系数均低于5%，与IDEXX试剂盒和RT-PCR的总体符合率分别为91.76%～93.33%和94.12%～96.67%。结果表明，基于2B10单克隆抗体建立的BVDV抗原捕获ELISA检测方法具有良好的特异性、敏感性和重复性，为BVDV的检测和防控提供借鉴技术手段。

关键词： 盖塔病毒   Cap蛋白   原核表达   多克隆抗体   免疫组织化学  

摘要： 为制备猪盖塔病毒（GETV）Cap蛋白多克隆抗体并探索其应用潜力，首先构建pCold-TF-Cap重组质粒，通过0.4 mmol/L IPTG诱导高效表达可溶性TF-Cap融合蛋白。经Ni-NTA亲和层析纯化后，利用SDS-PAGE鉴定重组蛋白纯度较高，BCA法测得蛋白浓度为22 mg/mL。以纯化的融合蛋白与佐剂乳化后免疫6～8周龄BALB/c小鼠，成功制备效价达1∶128 000的鼠源多克隆抗体。通过Western-blot分析与间接免疫荧光实验（IFA），证实该多克隆抗体能够特异性识别GETV感染的BHK-21细胞内源性Cap蛋白。此外，以制备的多克隆抗体为一抗、酶标羊抗鼠IgG为二抗，建立了检测GETV感染小鼠脾和肝组织中Cap蛋白的免疫组织化学（IHC）检测方法。结果显示，阳性信号清晰定位于细胞质中，呈现明显的棕色信号。本研究成功制备了特异性强的GETV Cap蛋白多克隆抗体，并建立了配套IHC检测方法，为GETV的基础研究和临床诊断提供了可靠的免疫学工具。

摘要： Constitutive JAK/STAT pathway activation is crucial in the pathogenesis of BCR::ABL1-negative myeloproliferative neoplasms (MPN), but has not yet been linked to interferon (IFN)-gamma signaling and tumor microenvironment. Human JAK2 V617F-mutated cell lines, 265 bone marrow biopsies (BMB) of two MPN cohorts, and 50 non-neoplastic BMB, revealed an intrinsic activation of IFN-gamma signaling, which was confirmed by public RNA expression data. In vitro analysis of JAK2-mutated cell lines showed an activation of IFN-gamma signaling pathway in the absence of IFN-gamma in the cell supernatants. In addition, a heterogeneous, but increased expression of IFN-gamma signaling components was found in BMB of JAK2-mutated samples with the highest expression in lymphocytes and monocytes, accompanied by increased tumor infiltrating lymphocytes (TIL). Unsupervised clustering identified a prognostic favorable cluster in both patient cohorts characterized by augmented IFN-gamma signaling and TILs. This cluster was enriched with JAK2-mutated, JAK-inhibition naive MPN, mainly essential thrombocythemia and polycythemia vera with mild bone marrow fibrosis. Moreover, in silico data confirmed the link between JAK2 mutations and increased IFN-gamma signaling. Multivariate Cox regression revealed TILs to be the strongest prognostic marker. In conclusion, JAK2-mutated MPN exhibit an intrinsic activation of IFN-gamma signaling associated with changes in the BM TME and patients' *** activation of the Janus kinases and signal transducer and activator of transcription (JAK/STAT) signaling pathway mainly mediated by mutations in the JAK2, CALR and MPL genes in pluripotent hematopoietic stem cells (HSC) is crucial for the pathogenesis of BCR::ABL1-negative myeloproliferative neoplasms (MPN). Despite the activation of JAK/STAT signaling and its influence on the proliferation of malignant cells is well studied in patient samples and JAK2- and CALR-mutated cell systems, there exists li

关键词： CS-PVA-IL/BaTiO   复合离子凝胶   挠曲电效应   柔性传感器   健康监测  

摘要： 传统压电器件难以有效感知关节活动引发的弯曲应变梯度，限制了其在可穿戴健康监测设备中的应用，因此亟需研发对弯曲形变响应灵敏的传感器以实现高效的人体动态信号感知。本文基于挠曲电效应，设计并制备了含有BaTiO3纳米颗粒的壳聚糖-聚乙烯醇-离子液体（CS–PVA–IL）复合离子凝胶，系统研究了BaTiO3含量对挠曲电响应特性的影响规律。当BaTiO3含量为0.25 wt%时，挠曲电响应电压高达1.8 ?V，挠曲电系数可达1.92 nC/m，展现出优异的挠曲电响应特性。所制备的传感器具有优异的线性响应特性与循环稳定性，在人体健康监测中，传感器在腕关节和食指弯曲过程中可产生清晰的响应电信号，响应电压峰值可有效反映弯曲幅度变化，验证了其在健康监测方面的实用性。本研究为柔性传感器的机电转换机制提供了新思路，为智能穿戴与生物电子领域的应用奠定了基础。