课程文献中心 更多>>

关键词： asymptomatic esophageal eosinophilia   brain-derived neurotrophic factor   EoE diagnostic panel   eosinophilic esophagitis   IL-13   iPSC-derived sensory neurons   neuropathic and inflammatory pain panel   neurotrophic receptor tyrosine kinase 2  

关键词： Leishmania   cytotoxicity   hypoxia   CD8 T cells   perforin  

摘要： Cutaneous leishmaniasis is a disease caused by protozoan parasites of the genus Leishmania , and although parasites influence disease severity, cytotoxic CD8 T cell responses mediate damage to the infected skin. We found that the cytotoxic protein perforin was expressed in CD8 T cells only upon recruitment to Leishmania -infected skin, suggesting that lesional inflammatory cues induced perforin. Here, using a mouse model of Leishmania major infection, we demonstrated that the expression of perforin was driven by a combination of hypoxia and IL-15, both of which are microenvironmental signals present within Leishmania -infected skin. We also demonstrated that the major sources of Il15 mRNA in cutaneous leishmaniasis lesions are neutrophils and macrophages and that macrophages exposed to hypoxia in vitro produce more Il15 . Since perforin is only present in lesions, we reformulated a small molecule perforin inhibitor for topical application and found that local inhibition of perforin is sufficient to ameliorate disease in established cutaneous leishmaniasis. Thus, topical perforin inhibition may be considered a therapeutic strategy for patients with cutaneous leishmaniasis and other inflammatory skin diseases where cytotoxic CD8 T cells contribute to disease pathogenesis.

关键词： IL-17A   COPD airway remodelling   Mouse bronchial fibroblasts   Autophagy  

摘要： The airway and lung tissue inflammation and structural changes caused by COPD lead to persistent and irreversible airflow limitation in patients. Several studies have associated IL-17A with COPD airway inflammation and collagen deposition, while autophagy is essential for maintaining normal cell function. Based on these findings, we propose a hypothesis that IL-17 affecting the autophagy of macrophages through the PI3K/AKT/mTOR pathway may contribute to the regulation of the airway remodeling process in COPD. The COPD airway remodelling model was confirmed by pulmonary function tests and HE and Masson staining. IL-17A, IL-6 and CCL20 were detected by ELISA, autophagosomes (APs) were observed by transmission electron microscopy, and western blotting was used to detect PI3K/AKT/mTOR-related proteins, autophagy-related proteins and collagen. Immunofluorescence revealed colocalization of LC3 in mouse bronchial fibroblasts (MBFs). MBFs were isolated and cultured via lentiviral transfection, IL-17RA overexpression or silencing, and quantitative analysis of PI3K/AKT/mTOR pathway-related proteins and autophagy-related proteins via western blotting. The results validated the establishment of the COPD model. Increased IL-17A in the COPD airway and increased IL-6 and CCL20 expression were observed in the COPD mice supplemented with the autophagy inhibitor 3MA. Using TEM, we observed a significant reduction in the number of APs in the airways in both the COPD and 3MA groups. Moreover, the PI3K/AKT/mTOR pathway phospho-p-PI3K/PI3K, p-AKT/AKT, and p-mTOR/mTOR ratios were also increased in the COPD group. Moreover, as P62 increased, Beclin-1 and LC3II/I expression decreased correspondingly, while Collagen I and Collagen III also increased. In our study of cultured mouse MBFs, the results showed that overexpression (OE) virus-mediated transfection of MBFs overexpressing IL-17RA increased the p-PI3K/PI3K, p-AKT/AKT, and p-mTOR/mTOR ratios, increased the P62 content and reduced the e

摘要： In unrelated allogeneic hematopoietic cell transplantation (allo-HCT), older and/or HLAmismatched donors are known risk factors for survival outcomes. In healthy individuals, cytomegalovirus (CMV) seropositivity is associated with impaired adaptive immune systems. We assessed whether the adverse effects of donor risk factors are influenced by the donor CMV serostatus. We analyzed 5836 patients with CMV seropositivity who received unrelated allo-HCT. We divided the entire cohort into 2 cohorts according to the donor CMV serostatus: CMV positive (DP) and negative (DN). We also stratified each cohort into 4 groups based on donor age (aged >= 40 or <40 years) and HLA parity (8/8 or 7/8): Young88 and Old88, and Young78 and Old78, respectively. In the CMV-DP cohort, the Old88 (hazard ratio [HR], 1.20;P = .012), Young78 (HR, 1.35;P < .001), and Old78 (HR, 1.60;P < .001) groups were associated with inferior overall survival (OS) than the Young88 group. In contrast, in the CMV-DN cohort, neither donor age nor HLA disparity was associated with inferior OS. The adverse impact of donor age was different between the cohorts (CMV-DP: HR, 1.19;P = .001;CMV-DN: HR, 1.04;P = .53;P for interaction, .070), as was the impact of HLA (CMV-DP: HR, 1.34;P < .001;CMV-DN: HR, 1.08;P = .23;P for interaction, .012). The impacts of donor age and HLA mismatch on OS might differ according to the donor CMV serostatus. In unrelated allo-HCT from a CMV-seronegative donor, an HLA-mismatched older donor may be able to be selected without affecting OS.

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（polymerase chain reaction， PCR）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*** 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和间接免疫荧光实验（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1:64 000。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。

关键词： 银屑病   克银合剂   凉血除湿法   IL-23/Th17细胞轴   临床疗效  

摘要： 目的：观察克银合剂治疗血热证中重度寻常型银屑病的临床疗效及安全性，初步探讨其对IL-23/辅助性T细胞17（Th17）细胞轴的干预作用。方法：收集门诊的血热证中重度寻常型银屑病患者30例作为研究组，选取健康体检的正常人30例作为对照组，研究组予克银合剂口服，30 mL/次，每日3次，疗程12周。比较研究组患者治疗前后的银屑病皮损面积与严重程度指数（psoriasis area and severity index，PASI）评分、体表皮损面积（body surface area，BSA）评分、皮肤病生活质量指数（dermatology life quality index，DLQI）评分，并比较治疗前两组间与研究组治疗前后血清中IL-17、IL-22、IL-23、IL-6、TNF-α的表达水平。结果：银屑病症状评分方面，治疗12周后，研究组患者的PASI评分、BSA评分、DLQI评分较治疗前均显著下降(P均＜0.01)，其中PASI 50及PASI 75的应答率分别为63.3%和16.7%。IL-23/Th17细胞轴相关因子方面，研究组患者的血清IL-17、IL-22、IL-6、IL-23、TNF-α水平显著高于健康对照组(均＜0.01)。研究组治疗12周后，患者的血清IL-22、IL-6、IL-17、IL-23、TNF-α 水平较治疗前均显著降低(P均＜0.01)。结论：克银合剂治疗血热证中重度寻常型银屑病疗效显著，安全性高，可以显著降低IL-23/Th17轴相关因子（IL-23、IL-22、IL-17、IL-6、TNF-α）水平。

摘要： PTP delta, encoded by PTPRD, is implicated in various neurological, psychiatric, and neurodevelopmental disorders, but the underlying mechanisms remain unclear. PTP delta trans-synaptically interacts with multiple postsynaptic adhesion molecules, which involves its extracellular alternatively spliced mini-exons, meA and meB. While PTP delta-meA functions have been studied in vivo, PTP delta-meB has not been studied. Here, we report that, unlike homozygous PTP delta-meA-mutant mice, homozygous PTP delta-meB-mutant (Ptprd-meB-/-) mice show markedly reduced early postnatal survival. Heterozygous Ptprd-meB+/- male mice show behavioral abnormalities and decreased excitatory synaptic density and transmission in dentate gyrus granule cells (DG-GCs). Proteomic analyses identify decreased postsynaptic density levels of IL1RAP, a known trans-synaptic partner of meB-containing PTP delta. Accordingly, IL1RAP-mutant mice show decreased excitatory synaptic transmission in DG-GCs. Ptprd-meB+/- DG interneurons with minimal IL1RAP expression show increased excitatory synaptic density and transmission. Therefore, PTP delta-meB is important for survival, synaptic, and behavioral phenotypes and regulates excitatory synapses in cell-type-specific and IL1RAP-dependent manners.

关键词： IL-28B   HTLV-1   HAM/TSP   HTLV-1 related diseases  

摘要： This research addresses IL-28B gene polymorphisms (rs12979860 and rs8099917) to determine their association with HTLV-1-related diseases;it aims to compare genotypic frequencies to identify predisposition or protection, considering population, disease, and controls. Given HTLV-1's impact on immunity, this study seeks biomarkers for early diagnosis and intervention. A systematic search met inclusion criteria, such as open access bibliographic and experimental studies published in English between 2010 and 2024, and genetic factors linked to susceptibility to pathologies. Regarding exclusion criteria, bibliographic or experimental studies in organisms other than humans, unofficial sources, non-indexed journals, and scientific articles in languages other than English were ruled out. Statistical data analyses were assessed using meta-analysis, including forest plot and Q test of heterogeneity based on the I2 statistics. The analyzed data indicate associations between genotypes, such as CT, GG, CC, and TT of the rs12979890 and rs8099917 polymorphisms and the predisposition to various diseases, such as HCV, arthropathy, HAM/TSP, cytomegalovirus and Crimean-Congo hemorrhagic fever associated with HTLV-1;however, the observed inconsistencies, such as high heterogeneity, and deficiency of related information limit the consolidation of the findings. Further research is needed to clarify IL-28B genotype interactions and disease susceptibility in HTLV-1 infections.

摘要： Intestinal epithelial-derived IL-18 is critical for homeostatic intestinal barrier function and is secreted through Gasdermin D (GSDMD) pores. Inflammasome activation is a prerequisite for both IL-18 maturation and GSDMD pore formation. However, GSDMD pores also cause pyroptotic cell death, which could be detrimental to the intestinal epithelial barrier. How epithelial cells balance the need to secrete IL-18 and to maintain barrier integrity remains poorly understood. In human intestinal epithelial cell lines and in primary human epithelial intestinal organoids, but not in immune cells, GSDMD plasma membrane pore formation by LPS electroporation and by gram-negative bacterial infection induced a non-conventional p37 caspase-4 fragment that was associated with reduced levels of mature IL-18. By contrast, limiting GSDMD plasma membrane pores pharmacologically and via point-mutagenesis prevented caspase-4 cleavage and increased IL-18 production, suggesting that p37 caspase-4 cleavage may regulate IL-18 maturation in the intestinal epithelium. In support, co-expression of caspase-4 cleavage mutants and IL-18 in HEK293T cells revealed that non-cleavable caspase-4 produced more mature IL-18 than cleaved caspase-4. Overall, these studies suggest that epithelial inflammasomes encode feedback pathways that control the balance between cytokine secretion and cell death. This may be an important mechanism to ensure homeostatic IL-18 production in the intestinal epithelium.

关键词： COVID-19   SARS-CoV-2   NL63   ACE2   IFN-beta   platform technology   fusion protein   type I interferon   flow cytometry   drug discovery   antivirals   emerging/re-emerging RNA viral pathogens   immunomodulators  

摘要： The COVID-19 pandemic highlighted the need for new therapeutic strategies to counter emerging pathogenic viruses. Herein, we introduce a novel fusion protein platform that enables antiviral targeting of distinct viral species based on host receptor specificity. Proof-of-concept studies focused on the human coronavirus NL63, which shares specificity for the ACE2 host receptor with the pandemic SARS-CoV and SARS-CoV-2 species. This antiviral fusion protein combines IFN-beta with the soluble extracellular domain of ACE2 (IFN beta-ACE2). Both domains retained predicted bioactivities in that the IFN-beta domain exhibited potent antiproliferative activity and the ACE2 domain exhibited full binding to the transmembrane SARS-CoV-2 Spike protein. In virus-washed (virus-targeted) and non-washed in vitro infection systems, we showed that the pool of IFN beta-ACE2 targeted to the virion surface had superior antiviral activity against NL63 compared to soluble ACE2, IFN-beta, or the unlinked combination of ACE2 and IFN-beta. The pool of IFN beta-ACE2 on the virion surface exhibited robust antiviral efficacy based on the preemptive targeting of antiviral IFN-beta activity to the proximal site of viral infection. In conclusion, virus-targeted IFN-beta places interferon optimally and antecedent to viral infection to constitute a new antiviral strategy.