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关键词： 系统性红斑狼疮   白细胞介素8   肽基精氨酸脱亚胺酶4   炎症   中性粒细胞   细胞因子  

摘要： 目的 探讨肽基精氨酸脱亚胺酶4（PADI4）对白细胞介素8（IL-8）分泌的调控，及其对系统性红斑狼疮（SLE）发病的影响。方法 利用qRT-PCR对比健康对照（HC）和SLE患者中性粒细胞IL-8表达水平。ELISA检测HC和SLE患者血清中IL-8分泌水平，分析狼疮患者血清IL-8水平与SLE相关血清学指标的关联性。ELISA检测HC或SLE血清对中性粒细胞分泌IL-8的影响。在原代中性粒细胞中使用PADI4特异性抑制剂GSK484，或在PADI4敲减的中性粒细胞样HL-60细胞（dHL-60）中检测N-甲酰-蛋氨酸-亮氨酸-苯丙氨酸（fMLP）及免疫复合物（ICs）刺激下IL-8的分泌水平。结果 与HC相比，IL-8高表达于SLE患者中性粒细胞。狼疮患者血清IL-8水平明显增高，且与血清IgM水平呈正相关。SLE患者血清能够诱导中性粒细胞分泌更多IL-8。PADI4抑制剂能够上调中性粒细胞中IL-8的产生，dHL-60细胞中，PADI4敲减导致IL-8分泌显著上升。结论 促炎细胞因子IL-8在SLE患者中性粒细胞和血清中高表达，并受PADI4调控，与狼疮血清学指标相关。IL-8通过炎症反应在SLE发生发展中发挥作用，PADI4/IL-8为SLE的监测及治疗提供新思路。

关键词： 鸡   MHC基因家族   生物信息学   系统进化   免疫应答  

摘要： 本研究旨在对鸡主要组织相容复合物（Major Histocompatibility Complex，MHC）基因家族进行生物信息学分析，明确鸡感染马立克氏病毒（Marek's Disease Virus， MDV）和注射羊红细胞（Sheep Red Blood Cell， SRBC）后MHC基因家族成员的表达模式。通过生物信息学工具构建跨物种系统进化树，分析鸡MHC蛋白的理化性质、亚细胞定位、染色体分布、保守结构域、共线性、顺式作用元件，并构建蛋白质互作网络，进行蛋白质三维结构建模。进一步分析鸡感染MDV和SRBC免疫应答后鸡脾脏中MHC基因的表达变化。研究共鉴定出49个鸡MHC基因家族成员，鸡MHC基因均密集分布于第16号染色体的特定区域，与日本鹌鹑亲缘关系最近。共线性分析发现其中存在2次串联重复和片段重复事件。蛋白特性分析发现MHC基因编码的蛋白具有1-2次跨膜结构域，均含有免疫球蛋白超家族保守结构域，且大多数主要定位于内质网和质膜。顺式元件分析发现MHC基因启动子区富含C/EBPα、NF-1等免疫相关调控位点，蛋白质互作网络分析发现BF2为枢纽节点。三维结构建模显示经典MHCⅠ闭合槽结构适配内源肽段呈递，Ⅱ类开放槽结构呈递外源肽段，而非经典基因（如MHCY）与BG基因可能通过非经典途径调控免疫应答。正选择分析发现鸟类特有的BG基因受到自然和人工的选择压力。转录组数据表明鸡感染MDV后大部分MHC基因的表达量上调。qRT-PCR分析表明鸡在SRBC免疫应答后，MHC Ⅱ基因（如BF1、BF2、BLB2和DMB2）表达显著上调。表明这些基因可能在免疫应答中起到重要作用。

关键词： cardiovascular diseases   clonal hematopoiesis   coronary disease   heart failure   ischemic stroke   venous thromboembolism   CHD   coronary heart disease   CHIP   clonal hematopoiesis of indeterminate potential   CVD   cardiovascular disease   HFpEF   heart failure with preserved ejection fraction   HFrEF   heart failure with reduced ejection fraction   IL   interleukin   smMIPS   single-molecule molecular inversion probe sequencing   VAF   variant allele frequency   VTE   venous thromboembolism  

摘要： Background Clonal hematopoiesis of indeterminate potential (CHIP) is an emerging aging-related risk factor for cardiovascular disease (CVD). However, previous studies suggest that CHIP’s relevance to CVD may diminish with advancing age. Objectives This study aimed to test the association of CHIP and its key subtypes with incident CVD in an older population. Methods Participants in the Women’s Health Initiative Long Life Study completed study assessments in 2012-2013 and underwent high-coverage sequencing (median depth 4,580×). The co-primary exposures were composite CHIP and TET2 CHIP. DNMT3A , ASXL1 , JAK2 , and non- DNMT3A CHIP were examined as secondary exposures. The primary outcome was incident coronary heart disease. Secondary outcomes were incident heart failure with preserved ejection fraction (HFpEF) and reduced ejection fraction (HFrEF), ischemic stroke, venous thromboembolism, and cardiovascular death. Multivariable-adjusted Cox models tested associations between CHIP and incident CVD. Results Among 6,677 women (median age 80 years; median follow-up 10.1 years), 2,176 (32.6%) had any CHIP. TET2 CHIP was independently associated with incident coronary heart disease (aHR: 1.36 [95% CI: 1.05-1.77]; P = 0.02), whereas composite CHIP was not (aHR: 1.07 [95% CI: 0.89-1.28]; P = 0.49). Secondarily, TET2 CHIP was associated with HFpEF (aHR: 1.40 [95% CI: 1.03-1.90]; P = 0.03), ASXL1 CHIP with HFrEF (aHR: 3.16 [95% CI: 1.53-6.55]; P = 0.002), and JAK2 CHIP with ischemic stroke (aHR: 2.49 [95% CI: 1.17-5.30]; P = 0.02), venous thromboembolism (aHR: 2.71 [95% CI: 1.11-6.65]; P = 0.03), and cardiovascular death (aHR: 2.62 [95% CI: 1.68-4.11]; P < 0.001). No other significant associations were observed for composite or DNMT3A CHIP. Conclusions In an older female cohort, key CHIP subtypes ( TET2 , ASXL1 , and JAK2 ) were associated with incident CVD, with associations that appeared to differ by CVD outcome. These findings suggest that CHIP remains associated with cardi

关键词： Feeder cells   NK cells expansion   mbIL-15   mbIL-21  

摘要： BACKGROUND:Natural killer (NK) cell immunotherapy is a promising approach for cancer treatment. However, its extensive clinical application was limited to the large-scale clinical-grade expansion of NK cells. In this study, we expanded NK cells from healthy donor's peripheral blood mononuclear cells (PBMCs) using a newly designed K562 feeder cell line. METHODS:The feeder cells were generated by transducing K562 cells with lentiviral particles carrying 4-1BBL and mbIL-21/-15. NK cells were expanded from PBMCs with these genetically modified, frozen-thawed and irradiated K562 feeder cells in the presence of IL-2. The purity, quantity, and receptors expression of the expanding NK cells were dynamically monitored. Furthermore, their anti-tumor efficacy was evaluated both in vitro and in vivo following a two-week expansion period. RESULTS:The K562-4-1BBL-mbIL-21/-15 feeder cells induced highly-efficient NK cells expansion from PBMCs (17902-fold) within two weeks. There was a notable upregulation in the expression of activating receptors including NKG2D, NKp30, NKp44, and NKp46 during the expansion process. Moreover, the expanded NK cells displayed enhanced cytotoxicity against a variety of hematological (K562, MOLM-13, OCI-AML-3, THP-1) and solid (Hep-G2, OVCAR3) cancer cell lines in vitro. In the humanized U937 xenograft mouse model, the NK cells extended the median survival time of the AML-bearing mice from 19.40 to 28.25 days. CONCLUSIONS:We have successfully established a highly-efficient, cost-effective and rapid NK cell expansion platform from PBMCs utilizing K562-4-1BBL-mbIL-21/-15 feeder cells, which also significantly improved the cytotoxicity both in vitro and in vivo, presenting a significant advancement in the field of NK cell-based immunotherapy.

关键词： CD8+ t cells   IL-24   Immunotherapy   Melanoma   Vinburnine  

摘要： BACKGROUND:Melanoma, a highly aggressive and immunogenic skin cancer, often develops resistance to immunotherapy due to the immunosuppressive tumor microenvironment (TME). Although PD-1/PD-L1 inhibitors have significantly improved treatment outcomes, 30%-40% of patients exhibit no response or develop resistance. Mechanisms such as T-cell exhaustion within the TME limit therapeutic efficacy, necessitating the exploration of novel strategies to enhance immune responses. METHODS:This study evaluated the effects of Vinburnine (Vin) on melanoma cell proliferation, migration, invasion, apoptosis, and DNA damage through in vitro experiments. Transcriptomic analysis, Western blot, RT-PCR, dual-luciferase reporter assays, and ChIP experiments revealed the mechanism by which Vin regulates IL-24 via ATF3. The anti-tumor efficacy of Vin or IL-24 in combination with PD-1 monoclonal antibody, as well as their modulation of the tumor microenvironment, were validated through luciferase-mediated cytotoxicity assays and a murine melanoma model. Additionally, the correlation between IL-24 expression and patient prognosis or immunotherapy response was analyzed using public databases. RESULTS:This study delineates the phenotypic mechanisms by which vinburnine suppresses melanoma proliferation. Vin induces reactive oxygen species (ROS) generation, leading to DNA damage and the subsequent activation of the apoptotic cascade in melanoma cells. Additionally, vinburnine activates the P38/MAPK/ATF3 signaling axis, which drives the secretion of interleukin-24 (IL-24), enhancing the functionality of CD8 T cells and modulating the tumor immune microenvironment to favor antitumor immunity. Notably, the combination of vinburnine with anti-PD-1 antibody therapy produces synergistic effects, effectively addressing certain limitations of current immunotherapeutic approaches. CONCLUSIONS:These findings underscore the therapeutic potential of vinburnine, particularly when used in combination with immun

摘要： Resident tissue macrophages and monocytes (RTMs) integrate local and systemic signals to coordinate immune cell function at homeostasis and in response to inflammatory stimuli. Obesity-associated metabolic dysfunction drives the development of RTM populations that contribute to disease states in multiple tissues. However, the contribution of specific dietary components to innate immune cell activation and function, as opposed to the direct effects of obesity, is largely unknown. Here, we studied the mechanisms by which high-fat (HF) diets shape lung RTM phenotype and function at steady state and influence responses to inflammatory insults. We found that, during HF diet feeding, lung RTMs accumulate saturated long-chain fatty acids, specifically stearic acid (SA), and demonstrate features of NLRP3 inflammasome priming and activation. In vivo, increased dietary SA was sufficient to cause neutrophil-predominant lung inflammation in the steady state and exacerbate a model of innate airway inflammation, whereas increased dietary oleic acid, the monounsaturated counterpart of SA, was sufficient to reduce inflammasome activation in the steady state and attenuate airway inflammation. Depletion of interleukin-1β (IL-1β) or pharmacologic inhibition of the endonuclease inositol requiring enzyme 1α (IRE1α) protected against SA-induced exacerbated lung inflammation. Last, we identified a population of lung monocytes with hallmarks of HF diet-induced RTM activation that were present in samples from obese humans with asthma. Together, these results identify a class of dietary lipids that regulate lung RTM phenotype and function in the steady state and modulate the severity of inflammation in the lung.

摘要： The oral mucosa exhibits unique regenerative properties and distinct wound healing dynamics compared to facial skin, providing a valuable model for studying tissue-specific repair mechanisms. Using a rigorously controlled mouse model combined with single-cell transcriptomics, we identified a novel mechanism underlying this tissue-specific difference. Our study revealed a population of fibroblast progenitors in the oral buccal mucosa that rapidly activate and differentiate into mature fibroblasts, contributing to effective wound resolution. Mechanistically, IL-1β derived from tissue-resident macrophages activates NFκB signals in these progenitors, inducing a metabolic shift from glycolysis to oxidative phosphorylation in a proteasome-dependent manner. This metabolic reprogramming supports stem cell differentiation and contributes to the unique regenerative pattern of the oral buccal mucosa. Our findings highlight the specialized healing mechanisms of the oral tissue and suggest that modulating proteasome activation and the IL-1β/NFκB axis may offer new therapeutic avenues for enhancing wound repair in tissue requiring extensive connective remodeling, such as skin or gingiva.

关键词： IL-1β   inflammasome   isoprenoids   leprosy   mevalonate pathway   prenylation