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关键词： Paraquat   IL-17A   Microglial phagocytosis   CD36   Amyloid-(3   alpha-Synuclein  

摘要： Paraquat (PQ), an environmental neurotoxin, has been demonstrated to induce pathological protein aggregation and thus neurotoxicity. Nevertheless, the exact mechanisms remain elusive. In this investigation, we explored the involvement of interleukin-17A (IL-17 A) in the aggregation of amyloid-(3 (A(3) and alpha-synuclein (alpha-syn) induced by PQ. Combining in vitro and in vivo, we explored whether PQ leads to A(3 and alpha-syn aggregation through IL-17 A-mediated reduction in microglia phagocytosis, thereby aggravating neurotoxicity. The results demonstrated that low-dose PQ continuous exposure significantly elevated IL-17 A levels in the peripheral blood serum and prefrontal cortical regions of mice. It also suppressed microglial phagocytosis of pathological proteins and promoted the aggregation of A(3 and alpha-syn in the prefrontal cortex. These changes ultimately resulted in depression, anxiety, and cognitive impairments. Mechanistically, IL-17 A inhibited the expression of the microglial phagocytic receptor CD36, impairing the microglial ability to clear A(3 and alpha-syn. Furthermore, administering an anti-IL-17 A effectively restored microglial phagocytosis in PQ-exposed mice, reduced A(3 and alpha-syn aggregation in prefrontal cortical areas, and alleviated behavioral deficits. In conclusion, this paper highlights IL-17 A as a pivotal mediator in PQ-induced neurotoxicity. It provides a potential target for developing novel therapeutic strategies against neurodegenerative pathologies induced by such environmental toxicants.

关键词： angiogenesis   early gastric cancer   endothelial cells   IL-33   tumor microenvironment   organoids   single-cell RNA sequencing  

摘要： Early gastric cancer (EGC) represents a critical stage in preventing and controlling the progression from gastritis to advanced gastric cancer (AGC). Therefore, identifying the single-cell characteristics of EGC, particularly the cellular composition of the tumor microenvironment (TME), as well as identifying potential predictive markers and therapeutic targets, could significantly enhance the monitoring of gastric cancer and improve clinical cure rates. We constructed a comprehensive single-cell RNA sequencing atlas for 184,426 high-quality gastric cancer cells from various stages, utilizing clinical biopsies and surgical samples. Our single-cell atlas highlights the cellular and molecular characteristics of EGC. Eight distinct cell lineage states were identified, and it was observed that the number of epithelial cell meta-clusters gradually decreased, while the number of T&NK, B, plasma, fibroblast, myeloid, and endothelial cells increased with disease progression. Certain epithelial subclusters (metaplastic stem-like cells (MSCs), pit mucous-like cells (PMC-like), proliferating cells), T-cell subclusters (Treg, CCR7+ naive, CH25H+ CD4+, TEM CD8+, and GFPT2+ CD8+ T cells), and endothelial subclusters (IL-33+ Venous-1 and AMAMTSL2+ Artery-2) were found to be increased in EGC. The Venous-1 subcluster was found to express high levels of IL-33. Mechanistically, it was revealed that IL-33 enhances the survival and angiogenesis of endothelial cells by upregulating the expression of adhesion proteins CD34 and PECAM1. Patient-derived EGC and AGC organoids were subsequently generated, and it was demonstrated that endothelial-derived IL-33 promoted the growth of both EGC and AGC organoids ex vitro and in vivo. Furthermore, IL-33 was found to increase the expression of KRT17 in EGC organoids. Notably, we also found that high expression of IL-33 was positively correlated with the depth of invasion and malignancy of EGC. This study provides novel insights into the single-cell

关键词： Aspergillus   IL-36   antifungal immunity   invasive pulmonary aspergillosis   macrophage   single nucleotide polymorphism   stem-cell transplantation  

摘要： BACKGROUND:Genetic predisposition plays a major role in the development of invasive pulmonary aspergillosis (IPA). The risk and course of IPA vary significantly among patients, yet continuously new genetic mechanisms that influence individual antifungal immune responses are being discovered. While genetic variability in IL-1 family cytokines is recognized as an important cause of disease susceptibility, it is unclear whether and how less-studied IL-1 family members, such as the IL-36 cytokine subfamily, are genetically regulated and influence the risk of infection. METHODS:We analyzed how genetic variants in the IL36 loci associate with the risk of IPA in 328 eligible recipients of allogeneic hematopoietic stem-cell transplants and their corresponding donors. The functional consequences of relevant genetic variants were investigated using clinical samples and in vitro infection models. RESULTS:We report that recipient, but not donor, SNPs rs895497 in IL36A and rs4849142 in IL36B increase the risk of IPA after transplantation. The strongest contribution of these SNPs to infection risk was observed in a combined analysis of transplant pairs. IL-36β was expressed in both human type II-like alveolar epithelial cells and macrophages following Aspergillus fumigatus infection. The risk genotype was associated with impaired production of IL-1β in bronchoalveolar lavage (BAL) samples from infected patients, as well as in fungal-stimulated macrophages. Moreover, macrophages harboring the risk genotype exhibited impaired fungicidal activity. CONCLUSIONS:Our findings suggest that genotype-specific mechanisms mediated by IL-36β act on the non-hematopoietic compartment, can impair antifungal immune responses in macrophages, and ultimately predispose to transplant recipient susceptibility to IPA.

关键词： Yellow fever virus   A549 cells   Mitochondrial network dynamics   Mitophagy   IFN-I   TLR2  

摘要： It has been emphasized that mitochondria play a fundamental role not only in cellular bioenergetics but also in the defense against infections. Here, we investigated mitochondrial network dynamics (MND) and IFN-I(3 signaling response in epithelial A549 cells after yellow fever virus (YFV) infection. We analyze the MND when only some cells are infected at 1 day post-infection (dpi) and after the spread of viral infection, at 3 dpi. Confocal microscopy and MiNA analysis showed that YFV infection leads to a decrease in the number of branches at 3 dpi and an increase in the length of branches at 1 and 3 dpi, suggesting that mitochondrial fission and fusion occur. Consistent with both processes, we found increased transcription of mitofusin 1 and Drp1 and increased colocalization of mitochondria with Drp1 at 3 dpi. In addition, mitochondrial membrane polarization decreased, mtROS production increased, p62 expression decreased, and LC3 expression increased, suggesting an increase in mitophagy flux. We found decreased expression of the IFN inducers RIG-I and MAVS sensors in YFV-infected A549 cells a t 3 dpi. Surprisingly, increased IFN-I(3 levels were observed at transcriptional and protein levels along with IRF7 induction at 1 and 3 dpi. Using the blocking antibody against TLR2, we showed that IFN-I(3 and IL-6 synthesis is maintained by TLR2 signaling. Mechanistically, infection led to activation of the NFxB pathway by degradation of IkBa, and increased phosphorylation of P65 and ERK MAPK signaling. Our results show that YFV infection induces altered MND in epithelial cells and triggers TLR2 signaling.

关键词： IL-17   IL-33   eosinophil   neutrophil  

关键词： 赤羽病病毒   Gn蛋白   昆虫杆状病毒表达系统   单克隆抗体  

摘要： 为制备针对赤羽病病毒（Akabane virus， AKAV）Gn蛋白的单克隆抗体，本研究将目的蛋白Gn的基因序列构建至pFastBac HTB载体，通过昆虫杆状病毒表达系统（BEVS）制备重组AKAV Gn蛋白。将纯化的重组蛋白免疫小鼠，取其脾细胞和骨髓瘤细胞（SP2/0）进行细胞融合及亚克隆，使用间接ELISA法筛选出能稳定分泌AKAV Gn蛋白的单克隆抗体（mAb）。经过Western Blot、IFA以及中和效价分析，证实获得的5株单抗均能分泌AKAV Gn蛋白，并能与AKAV感染的BHK-21细胞产生特异性反应，且中和效价均高于1︰27。单抗亚类鉴定结果表明5株单抗均为IgG亚类，轻链类型均为κ链。本试验制备的AKAV Gn单克隆抗体为深入探究AKAV Gn蛋白功能以及AKAV检测技术的完善奠定了基础。

关键词： β-乳球蛋白   超高压   白藜芦醇   结构   抗原性  

摘要： β-乳球蛋白（β-LG）是牛奶中的一种主要蛋白质，它可能会引起某些过敏体质人群的不适反应。本实验探究了不同压力下结合白藜芦醇（RES）对β-LG结构及抗原性变化的影响规律。通过光谱学技术分析其结构变化，而ELISA实验研究其抗原性变化。结果表明，不同压力条件下处理的β-LG的抗原性有不同程度的上升，但加入RES后其抗原性均发生下降。这是由于超高压处理对于β-LG的构象具有显著的影响，具体表现为α-螺旋、荧光强度、疏水性的上升，微观结构的破坏，从而暴露出了原本掩藏在内部的抗原表位，从而导致抗原性上升。而加入RES后，RES会与暴露出的抗原表位结合，使其α-螺旋、荧光强度下降，表面疏水性增加，从而破坏和修饰了致敏表位，使其抗原性降低。因此，超高压辅助多酚处理是一种有效的策略，能够通过改变β-LG的分子构象来降低其抗原性，为开发低抗原性牛奶产品提供了参考。

摘要： Osteoarthritis (OA) is a major global health problem with no disease-modifying therapies. Interleukin-1 (IL-1) is a critical cytokine associated with the pathophysiology of OA and can be inhibited by IL-1 receptor antagonist (IL-1Ra). Here, we tested the delivery of a gene therapeutic encoding the human IL-1Ra to the knee in a phase 1, open-label clinical trial that enrolled nine patients with radiographic knee OA. The IL-1Ra gene was delivered by a self-complementary (sc)-recombinant adeno-associated virus (rAAV) serotype 2.5 (sc-rAAV2.5IL-1Ra) by intra-articular injection into an index knee at one of three doses: low [1 x 1011 viral genomes (vg)], mid (1 x 1012 vg), or high (1 x 1013 vg). The primary outcome was safety. There were no serious adverse events (AEs) related to sc-rAAV2.5IL-1Ra. Two AEs occurred that were possibly related to the vector. Both were effusions with increased pain and resolved with conservative treatment. sc-rAAV2.5IL-1Ra did not cause changes in vital signs, physical findings, or clinical laboratory measures. Less than 1% of the injected dose of sc-rAAV2.5IL-1Ra vg was detected in circulation after 1 day and was cleared within a week. Titers of neutralizing antibodies to AAV2.5 rose in serum and synovial fluid. In all cases, IL-1Ra concentration increased in the synovial fluid, and IL-1Ra concentrations remained elevated after 1 year. Baseline pain and function scores improved during the study. Therefore, we found that intra-articular gene therapy with sc-rAAV2.5IL-1Ra was safe. The sustained increase in local IL-1Ra in human knee joints supports the further clinical examination of this therapy to provide therapeutic benefit in OA.