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摘要： TIM3, a T-cell inhibitory receptor, is expressed on exhausted T cells in the TME. Progressive loss of IL2-secretion is an early sign of diminished effector function in TILs, which raises the possibility of IL2 loss driving exhaustion of TILs. We show that endogenous IL-2 is required for the antitumor effect of anti-TIM3. Selective delivery of IL-2 to TIM3high TILs via an engineered anti-TIM3-Pro-IL2 fusion enhances anti-TIM3 efficacy, while reducing IL2 toxicity. IL2 activity is inhibited at the acidic pH of the TME, thus an IL2 mutein (IL2V2) with sustained activity at low pH is integrated into the construct. Mechanistically, TIM3-ProIL2V2 not only reactivates TIM3+ TILs but also facilitates the activation and expansion of TIM3- TILs, which in turn provide a sustained source of effector T cells. TIM3-ProIL2V2 is efficient in multiple tumor models, including tumors in humanized mice. TIM3-ProIL2V2 has the potential to overcome anti-PD-1/L1 resistance in cold cancers.

关键词： 细胞因子   流式细胞术   D-二聚体   病例报告  

摘要： 2024年2—11月该中心对3例细胞因子初检异常升高患者采用二步法进行了复测，3例患者初检时细胞因子均异常升高，复测后细胞因子水平显著降低。分析患者各项检测结果，发现3例患者体内D-二聚体水平均升高，疑似D-二聚体导致细胞因子检测假阳性，提示二者存在关联。

关键词： Herbicides   RAW 264.7   TNF   immunomodulatory activity   mitochondria   oxidative stress   clomazone   sulfentrazone   glyphosate  

摘要： Weeds are a concern in agriculture and the use of herbicides constitutes an effective, efficient, and economical way to control their growth. Recent discoveries of herbicides are promising for the management of resistant weeds. However, there is a gap in the knowledge of the toxic effects of some herbicides previously reported on immune cells. The present study aimed to examine cellular immunotoxicity of three herbicides (clomazone, glyphosate, and sulfentrazone) after 96 hr incubation utilizing RAW 264.7 BALB/c mouse monocyte/macrophage-like cell line to elucidate the role of some toxicological pathways. Data demonstrated the herbicides clomazone, glyphosate, and sulfentrazone initiated a cytotoxic effect as evidenced by EC50 values of 429.2;53.7;866.6 mg/L, respectively. Clomazone and sulfentrazone, at all concentrations, induced excess production of reactive oxygen (ROS) and reactive nitrogen (RNS) free radicals. An immunosuppression was observed in RAW 264.7 cells after incubation with 50 or 100 mg/L glyphosate and 500 or 1000 mg/L sulfentrazone. In addition, all herbicides produced mitochondrial depolarization and decreased tumor necrosis factor-alpha (TNF-alpha) levels. This constitutes the first report of the effects of clomazone and sulfentrazone on RAW 264.7 cells, including reduced TNF-alpha levels, indicating the adverse influence of herbicides on the immune system.

关键词： 肾衰老   S100钙结合蛋白A8/9   NLRP3/Caspase-1/IL-1β通路   慢性炎症  

摘要： 目的 探讨S100A8/9及介导的NLRP3/Caspase-1/IL-1β通路在由D-gal诱导的大鼠肾衰老模型和H2O2诱导的HK-2细胞衰老模型中的作用。方法 12只SD大鼠随机分为正常组和D-gal组，经颈背部皮下注射D-gal（150mg/kg）建立大鼠肾衰老模型，8周后麻醉状态下收集肾标本，荧光定量PCR检测衰老相关基因p21，p16和p53mRNA表达水平；SA-β-Gal染色观察肾组织衰老情况，HE和Masson染色观察肾组织病理学变化；检测血清中BUN和CREA含量，检测肾组织中SOD、CAT、GSH-PX和MDA活性，DHE染色观察肾组织ROS水平变化；免疫荧光染色检测S100A8/9蛋白表达水平，Westernblot检测纤维化标志物CollagenⅢ、α-SMA和TGF-β1以及NLRP3/Caspase-1/IL-1β通路上相关蛋白的表达水平。体外利用H2O2构建HK-2细胞肾衰老模型，检测衰老蛋白p21、p16表达水平和炎症因子IL-18、TNF-αmRNA表达水平，SA-β-Gal染色观察细胞衰老情况；并使用S100A8/9抑制剂帕奎莫德（paquinimod，PAQ）干预，检测S100A8/9及NLRP3/Caspase-1/IL-1β通路上相关蛋白的表达水平。结果 与正常组大鼠比较，D-gal组大鼠肾组织衰老基因p21，p16和p53mRNA表达水平显著增加（P ＜0.01），衰老相关β-半乳糖苷酶（senescence-associatedβ-galactosidase，SA-β-Gal）染色显示衰老细胞数量显著增多（P ＜0.01）；血清中BUN和CREA含量上升（P ＜0.05）；组织内CAT、GSH-PX和SOD活性显著下降（P ＜0.01），MDA活性显著增加（P ＜0.01）；纤维化标志物CollagenⅢ、αSMA和TGF-β1的蛋白表达增加（P ＜0.05）；组织内ROS含量增多（P ＜0.05）；D-gal组大鼠肾小球出现一定程度的萎缩及缺失，肾小囊囊腔和肾小管管腔增大，细胞核深染、固缩，出现大量胶原纤维沉积；S100A8和S100A9蛋白表达增加（P ＜0.01）；NLRP3/Caspase-1/IL-1β信号通路上NLRP3、Caspase-1和IL-1β蛋白表达增加（P ＜0.05）。H2O2诱导HK-2细胞衰老后，使用S100A8/9抑制剂PAQ减轻了HK-2细胞衰老情况，衰老蛋白p21、p16表达水平和炎症因子IL-18、TNF-αmRNA表达水平均降低（P ＜0.05，P ＜0.01），SA-β-Gal染色显示衰老细胞数量减少（P ＜0.01），并抑制S100A8和S100A9蛋白表达（P ＜0.01）以及NLRP3、Caspase-1和IL-1β蛋白的过表达（P ＜0.05或P ＜0.01）。结论 S100A8/9通过激活NLRP3/Caspase-1/IL-1β通路参与慢性炎症反应，从而促进D-gal诱导的肾衰老过程。

关键词： 慢性胃炎   胆汁反流   脾胃湿热证   黄连温胆汤   临床研究  

摘要： 目的：观察黄连温胆汤加减治疗脾胃湿热型慢性胃炎伴胆汁反流的临床疗效及其对肿瘤坏死因子-α（TNF-α）、白细胞介素-8（IL-8）、前列腺素E2（PGE2）和表皮生长因子（EGF）的影响。方法：选择符合纳入标准的慢性胃炎伴胆汁反流脾胃湿热证患者82例，依据随机数字表随机分为观察组和对照组，每组41例，治疗过程中，观察组脱离及剔除3例，对照组脱离及剔除2例，最终观察组38例，对照组39例。对照组给予铝碳酸镁咀嚼片，观察组给予黄连温胆汤加减口服，两组均治疗4周。比较两组患者治疗前后中医症状评分及证候总评分、TNF-α、IL-8、PGE2、EGF含量的变化情况，及临床疗效和复发率。结果：观察组治疗总有效率为94.74%（36/38），高于对照组82.05%（32/39）(P <0.05)。治疗后，两组中医各症状评分及证候总评分均显著低于治疗前（P <0.05,P <0.01），且观察组改善情况优于对照组（P <0.05,P <0.01）。与治疗前比较，治疗后两组患者TNF-α、IL-8含量均显著降低（P <0.01）,PGE2、EGF含量均显著升高（P <0.01）；治疗后组间比较，观察组TNF-α、IL-8均低于对照组（P <0.01）,PGE2、EGF均显著高于对照组（P <0.01）。治疗结束6个月后随访，观察组复发率为8.33%（3/36），对照组复发率为12.5%（4/32），观察组低于对照组，但差异无统计学意义。两组患者在治疗期间均未发生不良反应及毒副作用。结论：黄连温胆汤加减治疗慢性胃炎伴胆汁反流脾胃湿热证临床疗效显著，其作用机制可能与调节血清TNF-α、IL-8、PGE2、EGF表达水平有关。

摘要： Dysregulations in protein kinases significantly contribute to the initiation, progression, and drug resistance in non-small cell lung cancer (NSCLC). Identification of novel oncogenic drivers within the human kinome is crucial for targeted therapy. In this study, we conducted a comprehensive analysis of the TCGA database and literature, pinpointing 16 candidate genes in lung cancer exhibiting frequent dysregulation and limited research. Our functional analysis revealed Serine/threonine kinase 31 (STK31) as a key player in driving tumor growth, in immune-competent mice, with minimal impact in nude mice. Further investigations unveiled upregulation of STK31 led to CD8+ T cell exhaustion. Mechanistically, STK31 induced CD8+ T cell exhaustion through the signal transducer and activator of transcription 3 (STAT3) - interleukin 6 (IL-6) signaling pathway. Direct interaction between STK31 and STAT3 activated the transcription of downstream oncogenic targets, such as IL-6, facilitating immune escape. Moreover, STK31 exhibited elevated expression levels in lung cancer tissues compared to adjacent tissues and displayed a significant correlation with poor prognosis in lung cancer patients. This study defines a critical role of STK31 in promoting immune escape through STAT3 activation, positioning it as a promising therapeutic target for lung cancer.

关键词： Glycosylation   Autoimmune disorder   Homology modeling   Molecular docking   Molecular dynamics simulation  

摘要： Interleukin-1 receptor antagonist (IL-1ra) is a vital anti-inflammatory molecule, acting as a decoy protein that competitively binds to Interleukin-1 receptors (IL-1RI and IL-1RII). The deregulation of IL-1 activity has been linked to autoimmune disorders, impaired wound healing, and the occurrence of cytokine storms in the context of infections. At present, *** expressed IL-1ra isoform was used as a therapeutic drug for the treatment of IL-1 mediated inflammatory diseases;however, daily dosing raises concerns about allergens and infection. Notably, the drug lacks glycosylation, an important post-translational modification for protein stability and pharmacodynamics. The present study utilizes computational molecular dynamics simulations to investigate the influence of glycosylation on the stability and binding efficacy of IL-1ra. To study the impact, we introduced mammalian bi-antennary hybrid glycan with terminal sialic acid/ N-Acetylneuraminic acid (Neu5Ac) added to the respective Asparagine (Asn) residue of 3D modeled IL-1ra structure. The molecular dynamics simulations demonstrated that glycosylated IL-1ra increased the structural stability and exhibited a sustained level of binding stability with 24 hydrogen bonds and 8 salt bridges in comparison to non-glycosylated IL-1ra protein with 20 hydrogen bonds and 6 salt bridges as the result of post simulation. This finding underscores the possibility of glycosylation as it means to augment the effectiveness of therapeutic medications that specifically target immune regulation. These understandings help to translate the concept to the complete expression of proteins. Our study addressed the mechanism of how the N-glycan regulates cytokine, which will provide insight into the design of novel anti-inflammatory drugs and specifically glycan play a role in RA.

关键词： Brain death   TNF-alpha   kidney donor  

摘要： Kidney transplant outcomes depend on various factors, including donor type. Brain death (BD) has been associated with significant kidney injury, with long-term sequelae. Deciphering the impact of BD circulatory death on kidney pathophysiology may help to prevent kidney transplant injury. We developed 3 rat models: 6h of BD (DBD, n=10); circulatory death (DCD, n=10) with 20min of warm ischemia; and 6-hour BD with renal denervation prior to BD (DNRV, n=6). RNA-seq was performed on kidney tissue, followed by differential gene expression and pathway enrichment analyses. Circulating cytokines were measured by Luminex/ELISA. Our findings were validated in 10 human pre-implantation biopsies matching our rat models (n=5 DBD; n=5 DCD). After BD, serum creatinine levels increased from 0.33 [0.31-0.38] to 0.61 [0.59-0.67]mg/dL (p<0.0001). Compared to DCD, DBD rats exhibited significantly higher levels of IL-6 (26.67 [19.10-32.35] 10.32 [9.76-13.28]ng/mL, p=0.006) and TNFᵬ (21.98 [16.48-32.60] 9.64pg/mL [7.96-11.13], p=0.0012), and lower levels of IL-10 (1.28 [1.11-1.65] 1.79 [1.46-2.18]ng/mL, p=0.049). Differential expression analysis revealed that DBD kidneys exhibited CD11b-associated inflammation, caspase-3-mediated apoptosis, and TNFᵬ-driven injury (p=7.34×10; z-score=2.475), while DCD kidneys activated EIF2/NRF2 stress-response pathways. Notably, denervation did not mitigate BD-induced injury. Human biopsies confirmed the positive enrichment of inflammatory pathways (p=0.01 to 1.18×10) in DBD compared to DCD, with 6 out of the top 15 pathways shared across species. Our findings highlight specific mechanisms affecting differently kidneys derived from DBD DCD donors, with a potential role of TNFᵬ in BD-associated kidney injury. Kidney denervation prior to BD does not prevent kidney injury.

关键词： Tripterygium wilfordii polyglycoside tablets   Hepatocellular carcinoma   therapeutic drug   IL-6   PI3K/MAPK/STAT3 signaling pathway  

摘要： Tripterygium wilfordii polyglycoside tablets (TWPT) have traditionally been used to treat certain inflammatory diseases. This study validated TWPT as a novel application in hepatocellular carcinoma treatment through multiple targets, thereby expanding its clinical medication scope. TWPT exhibited a low toxicity and a significantly antihepatoma effects in vitro and in vivo. Through network pharmacology analysis, we found TWPT attenuated the progression of hepatocellular carcinoma by multi-targeting, including IL-6, MMP9, TNF-alpha and VEGFA. Additionally, TWPT targeted IL-6 to regulate downstream pathways, including the PI3K/Akt, JAK2/STAT3, and MAPK signaling pathways. Thus, TWPT could be developed as a potential therapeutic drug for hepatocellular carcinoma.

关键词： Porcine pancreatic elastase (PPE)   Emphysema   Siglec-F+ neutrophils   IL-17A   G-CSF   gamma delta(+) T cells   Epithelial/stromal cells   G-CSF receptor signaling  

摘要： Neutrophils play a pivotal role in the progression of IL-17-mediated airway inflammation, but the mechanisms underlying their pathological differentiation remain poorly understood. In this study, we identified a distinct population of lung-specific pathogenic Siglec-F+ neutrophils in a porcine pancreatic elastase (PPE)-induced mouse model of emphysema. Compared with conventional neutrophils, these Siglec-F+ neutrophils exhibited increased phagocytic activity, increased extracellular trap formation, increased production of proinflammatory cytokines, and reduced IL-10 levels. During the early phase of acute inflammation following PPE instillation, IL-17A levels in the lungs increase, which is driven primarily by gamma delta(+) T cells. IL-17A stimulated lung epithelial/stromal cells to secrete granulocyte colony-stimulating factor (G-CSF), which promoted the differentiation of Siglec-F+ neutrophils via the JAK2/STAT3 pathway and the PI3K-independent mTOR and p38 MAPK signaling pathways. Neutralizing G-CSF or inhibiting JAK2/STAT3, mTOR or p38 MAPK signaling significantly suppressed Siglec-F+ neutrophil development, resulting in the alleviation of emphysematous symptoms. Our findings underscore the crucial role of Siglec-F+ neutrophils in the pathogenesis of PPE-induced emphysema and suggest that targeting the IL-17A/G-CSF axis or G-CSF receptor downstream signaling pathways may represent a promising therapeutic strategy for treating emphysema.