Cellular immunology最新文献_第9页

Nebulized mesenchymal stem cell-derived exosomes attenuate airway inflammation in a rat model of chronic obstructive pulmonary disease 雾化间充质干细胞衍生的外泌体减轻慢性阻塞性肺疾病大鼠模型的气道炎症

IF 3.7 4区医学 Q2 CELL BIOLOGY

Cellular immunology

Pub Date : 2025-03-01 DOI: 10.1016/j.cellimm.2025.104933

Min Wang , Yuxin Hao , Wei He , Hui Jia , Zhaoshuang Zhong , Shuyue Xia

Chronic Obstructive Pulmonary Disease (COPD) is one of the leading causes of death worldwide, and current treatments fail to significantly halt its progression. Exosomes derived from mesenchymal stem cells (MSCs-Exos) have demonstrated promising potential in treating COPD due to their anti-inflammatory and regenerative biological properties. In this study, we investigated the potential anti-inflammatory effects of bone marrow mesenchymal stem cell-derived exosomes (BMSCs-Exos) in a COPD rat model and the possible mechanisms by which they inhibit airway remodeling, as well as identifying the optimal dosage and administration route. Our results show that nebulized BMSC-Exos significantly improve lung function in COPD rats while reducing pulmonary inflammatory infiltration, bronchial mucus secretion, and collagen deposition. Moreover, BMSC-Exos treatment notably decreased the expression of pro-inflammatory cytokines such as TNF-α, IL-6 and IL-1β, and the pro-fibrotic factor TGF-β1 in serum, bronchoalveolar lavage fluid (BALF), and lung tissue. The most pronounced therapeutic effect was observed at a low dose of exosomes. Furthermore, quantitative real-time PCR and immunohistochemical analyses revealed that nebulized BMSC-Exos significantly inhibited airway remodeling and epithelial–mesenchymal transition (EMT) by suppressing the Wnt/β-catenin signaling pathway. In conclusion, these findings indicate that nebulized BMSC-Exos offer a noninvasive therapeutic strategy for COPD by mitigating lung inflammation and airway remodeling through the suppression of abnormal Wnt/β-catenin pathway activation induced by cigarette smoke (CS) and lipopolysaccharide (LPS) in rats.

慢性阻塞性肺疾病（COPD）是世界范围内导致死亡的主要原因之一，目前的治疗方法未能显著阻止其进展。来自间充质干细胞（msc - exos）的外泌体由于其抗炎和再生的生物学特性，在治疗COPD方面显示出很大的潜力。在这项研究中，我们研究了骨髓间充质干细胞衍生外泌体（BMSCs-Exos）在COPD大鼠模型中的潜在抗炎作用，以及它们抑制气道重塑的可能机制，并确定了最佳剂量和给药途径。我们的研究结果表明，雾化BMSC-Exos可显著改善COPD大鼠的肺功能，同时减少肺部炎症浸润、支气管粘液分泌和胶原沉积。此外，BMSC-Exos处理显著降低血清、支气管肺泡灌洗液（BALF）和肺组织中促炎因子TNF-α、IL-6、IL-1β和促纤维化因子TGF-β1的表达。在低剂量的外泌体中观察到最显著的治疗效果。此外，定量实时PCR和免疫组织化学分析显示，雾化BMSC-Exos通过抑制Wnt/β-catenin信号通路，显著抑制气道重塑和上皮-间质转化（EMT）。综上所述，这些研究结果表明，雾化BMSC-Exos通过抑制香烟烟雾（CS）和脂多糖（LPS）诱导的大鼠Wnt/β-catenin通路异常激活，减轻肺部炎症和气道重塑，为慢性阻塞性肺疾病提供了一种无创治疗策略。

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引用次数: 0

Breast cancer stem cells convert anti-tumor CD4+ T cells to pro-tumor T regulatory cells: Potential role of exosomal FOXP3 乳腺癌干细胞将抗肿瘤CD4+ T细胞转化为促肿瘤T调节细胞：外泌体FOXP3的潜在作用

IF 3.7 4区医学 Q2 CELL BIOLOGY

Cellular immunology

Pub Date : 2025-02-20 DOI: 10.1016/j.cellimm.2025.104931

Udit Basak , Sourio Chakraborty , Sumon Mukherjee , Subhadip Pati , Poulami Khan , Subhajit Ghosh , Arghya Adhikary , Kuladip Jana , Gaurisankar Sa , Tanya Das

Cancer progression and its treatment-response are regulated by the tumor microenvironment (TME). Tumor-initiating cancer stem cells (CSCs) remain in constant communication with the TME, and modulate it through several mechanisms. Here, from in-silico findings and analyzing breast cancer patient tissue-derived data, CSCs and Tregs were found to be positively correlated. Furthermore, our in-silico analyses highlighted a positive relationship between CSC genes and Treg signature marker, FOXP3, even in cancer cell lines that do not contain any T cell or Treg cells, thus raising the possibility of CSCs expressing FOXP3. Validating our hypothesis, higher expression of FOXP3, both at mRNA and protein levels, was observed in breast CSCs than non-stem cancer cells. Since a small population of CSCs initiate tumor in immune cell-dominated TME, we aimed at exploring whether breast CSCs directly transfer FOXP3 to CD4⁺T cells to generate immunosuppressive Treg cells. First, our search revealed CSC-derived exosomes (CDEs) generated CD4⁺CD25⁺FOXP3⁺ Tregs at an early time-point of 24 h, which were immunosuppressive in nature. Next, detecting presence of FOXP3 protein in CDEs showed a strong possibility of FOXP3 transfer through CDEs. This was supported by detecting elevated FOXP3 levels from 12 h in translation inhibitor-treated T cells upon CDE-exposure. Finally, exosomes derived from FOXP3 attenuated-CSCs furnished lower FOXP3 in T cells than control CDEs. This mechanism was validated in in-vivo murine model. Together these results indicate a hitherto unexplored role of CSC-derived FOXP3 in reprogramming T cells into immunosuppressive Treg cells.

肿瘤的进展及其治疗反应受肿瘤微环境（tumor microenvironment， TME）的调控。肿瘤启动性癌症干细胞（CSCs）与TME保持着持续的交流，并通过几种机制对其进行调节。在这里，从计算机发现和分析乳腺癌患者组织来源的数据，发现CSCs和Tregs呈正相关。此外，我们的计算机分析强调了CSC基因与Treg标记物FOXP3之间的正相关，即使在不含任何T细胞或Treg细胞的癌细胞系中也是如此，从而提高了CSC表达FOXP3的可能性。验证我们的假设，在乳腺CSCs中观察到FOXP3在mRNA和蛋白水平上的表达高于非干细胞。由于一小群CSCs在免疫细胞主导的TME中引发肿瘤，我们旨在探索乳腺CSCs是否直接将FOXP3转移到CD4+T细胞中以产生免疫抑制的Treg细胞。首先，我们的研究发现csc衍生的外泌体（CDEs）在24小时的早期时间点产生CD4+CD25+FOXP3+ treg，其本质上具有免疫抑制作用。接下来，检测CDEs中FOXP3蛋白的存在，表明FOXP3通过CDEs转移的可能性很大。在翻译抑制剂处理的T细胞暴露于cde后12小时检测到FOXP3水平升高，这一点得到了支持。最后，来自FOXP3减弱的cscs的外泌体在T细胞中提供的FOXP3低于对照CDEs。该机制在小鼠体内模型中得到了验证。综上所述，这些结果表明csc来源的FOXP3在将T细胞重编程为免疫抑制性Treg细胞中的作用迄今尚未被探索。

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引用次数: 0

In vitro ovarian tumor-conditioned CD163+ human macrophages retain phagocytic response to CD47 blockade 体外卵巢肿瘤条件CD163+人巨噬细胞对CD47阻断保持吞噬反应

IF 3.7 4区医学 Q2 CELL BIOLOGY

Cellular immunology

Pub Date : 2025-02-17 DOI: 10.1016/j.cellimm.2025.104932

Kristian W. Antonsen , Anne G. Jensen , Boe S. Sorensen , Anders Etzerodt , Søren K. Moestrup , Holger J. Møller

Introduction

CD163-expressing macrophages are abundant in ovarian cancer where they accelerate tumor growth and metastasis. CD47 blockade is a novel immunotherapy aiming to activate macrophage phagocytosis of tumor cells, but it is currently unknown if the tumor-associated macrophages expressing CD163 respond poorly to CD47 blockade.

Methods

Human monocyte-derived macrophages were exposed to tumor-conditioned medium from A2780 ovarian cancer cells during differentiation. Effects on gene expression, membrane protein levels, release of soluble proteins and macrophage phagocytosis of A2780 cells in response to CD47 blockade were measured and compared to control macrophages.

Results

Tumor cell conditioning induced macrophage expression of CD163 on both the mRNA and protein level. Furthermore, tumor conditioning simultaneously increased protein expression of the phenotype markers CD206 and CD80, and the phagocytosis checkpoint LILRB1. However, tumor conditioning did not reduce phagocytic capacity, as CD47 blockade induced macrophage phagocytosis of A2780 cells to similar degrees in both control and tumor cell-conditioned macrophages.

Discussion

In vitro tumor conditioning did not reduce the phagocytic response to CD47 blockade, suggesting that induction of a macrophage phenotype with increased expression of CD163 does not directly limit the capacity for phagocytosis of tumor cells. In conclusion, these findings suggest that CD163+ macrophages remain responsive to CD47 blockade, highlighting their potential as targets for immunotherapy in ovarian cancer.

表达cd163的巨噬细胞在卵巢癌中大量存在，它们加速肿瘤的生长和转移。CD47阻断是一种新的免疫疗法，旨在激活巨噬细胞吞噬肿瘤细胞，但目前尚不清楚表达CD163的肿瘤相关巨噬细胞是否对CD47阻断反应不良。方法将人单核细胞来源的巨噬细胞暴露于A2780卵巢癌细胞分化过程中的肿瘤条件培养基中。检测CD47阻断对A2780细胞基因表达、膜蛋白水平、可溶性蛋白释放和巨噬细胞吞噬的影响，并与对照巨噬细胞进行比较。结果肿瘤细胞调节诱导巨噬细胞在mRNA和蛋白水平上表达CD163。此外，肿瘤调节同时增加了表型标记CD206和CD80以及吞噬检查点LILRB1的蛋白表达。然而，肿瘤调节并没有降低吞噬能力，因为CD47阻断在对照和肿瘤细胞条件的巨噬细胞中诱导A2780细胞的吞噬程度相似。体外肿瘤调节并没有降低对CD47阻断的吞噬反应，这表明诱导巨噬细胞表型增加CD163的表达并不直接限制肿瘤细胞的吞噬能力。总之，这些发现表明CD163+巨噬细胞仍然对CD47阻断反应，突出了它们作为卵巢癌免疫治疗靶点的潜力。

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引用次数: 0

The polarization of macrophages participates in the repair after folic acid-induced acute kidney injury 巨噬细胞极化参与叶酸诱导急性肾损伤后的修复

IF 3.7 4区医学 Q2 CELL BIOLOGY

Cellular immunology

Pub Date : 2025-02-10 DOI: 10.1016/j.cellimm.2025.104929

Shujie Yang , Yan Shen

Acute kidney injury (AKI) remains a major public health challenge, posing serious threats to human health. Increasing evidence indicates that renal cells undergo significant metabolic alterations following AKI, with inflammatory responses persisting throughout both injury and repair phases. Our previous research has demonstrated that heightened aerobic glycolysis after AKI leads to increased secretion of metabolic byproducts such as lactate, which plays a critical role in tissue repair. However, the relationship between metabolic reprogramming and inflammatory responses, as well as the underlying mechanisms, remain poorly understood. This study aims to clarify the regulatory effects of the glycolytic byproduct lactate on macrophage activation and phenotypic differentiation following AKI. We observed increased expression of M1/M2 macrophages and elevated secretion of inflammatory cytokines after folic acid-induced AKI. Immunofluorescence staining showed co-localization of macrophages with α-SMA. Manipulating lactate levels post-injury led to a decrease in macrophage expression and a reduction in fibroblast activation and proliferation, ultimately impairing renal tissue repair. These findings suggest that targeting lactate as a key regulator of macrophage phenotype differentiation may provide a theoretical and clinical foundation for therapeutic strategies in AKI repair.

急性肾损伤（AKI）仍然是一项重大的公共卫生挑战，对人类健康构成严重威胁。越来越多的证据表明，肾细胞在AKI后发生了显著的代谢改变，炎症反应在损伤和修复阶段都持续存在。我们之前的研究表明，AKI后有氧糖酵解的增加导致代谢副产物如乳酸的分泌增加，乳酸在组织修复中起着关键作用。然而，代谢重编程和炎症反应之间的关系以及潜在的机制仍然知之甚少。本研究旨在阐明糖酵解副产物乳酸对AKI后巨噬细胞活化和表型分化的调节作用。我们观察到叶酸诱导AKI后M1/M2巨噬细胞表达增加，炎症细胞因子分泌升高。免疫荧光染色显示巨噬细胞与α-SMA共定位。损伤后乳酸水平的改变导致巨噬细胞表达减少，成纤维细胞活化和增殖减少，最终损害肾组织修复。这些发现表明，将乳酸作为巨噬细胞表型分化的关键调节因子，可能为AKI修复的治疗策略提供理论和临床基础。

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引用次数: 0

Tim-3 promotes viral infection by suppressing the USP25-TRAF3-IRF7 signaling pathway Tim-3通过抑制USP25-TRAF3-IRF7信号通路促进病毒感染

IF 3.7 4区医学 Q2 CELL BIOLOGY

Cellular immunology

Pub Date : 2025-02-08 DOI: 10.1016/j.cellimm.2025.104930

Lin Du , Jinjie Chen , Chunxiao Du , Junrui Chen , Zhaoxiang Wang , Bing Bao , L.V. Zhonglin , Chen Xing , Meng Liang , Lanying Wang , Shun Xie , Yuxiang Li , Zhiding Wang , Ge Li , Jun Zhang , Gencheng Han

Tim-3, an immune checkpoint inhibitor, plays key roles in maintaining immune homeostasis and is involved in viral evasion. However, the precise role of Tim-3 in viral infection remains to be determined. USP25 is a deubiquitinating enzyme that initiates antiviral immunity by deubiquitinating TRAF3 and triggering the antiviral signaling pathway. Here we found that Tim-3-specific knockout in myeloid cells leads to enhanced antiviral immunity in mice with vesicular stomatitis virus (VSV) encephalitis by increasing the type I interferon response. Mechanistically, Tim-3 inhibits the expression of USP25 via STAT1 and interacts with USP25 but does not regulate its posttranslational modification; as a result, Tim-3 inhibits USP25-mediated deubiquitination of TRAF3, promotes K48-linked ubiquitination and degradation of TRAF3, inhibits the phosphorylation of IRF7, and ultimately downregulates the interferon response. These findings provide new insights into the function of Tim-3 in antiviral immunity and its related clinical significance.

Tim-3是一种免疫检查点抑制剂，在维持免疫稳态中起关键作用，并参与病毒逃避。然而，Tim-3在病毒感染中的确切作用仍有待确定。USP25是一种去泛素化酶，通过去泛素化TRAF3和触发抗病毒信号通路来启动抗病毒免疫。在这里，我们发现髓细胞中tim -3特异性敲除通过增加I型干扰素应答，导致水疱性口炎病毒（VSV）脑炎小鼠抗病毒免疫增强。在机制上，Tim-3通过STAT1抑制USP25的表达，并与USP25相互作用，但不调节其翻译后修饰；因此，Tim-3抑制usp25介导的TRAF3去泛素化，促进k48相关的TRAF3泛素化和降解，抑制IRF7的磷酸化，最终下调干扰素应答。这些发现为Tim-3在抗病毒免疫中的功能及其临床意义提供了新的认识。

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引用次数: 0

Sodium butyrate prevents lipopolysaccharide induced inflammation and restores the expression of tight junction protein in human epithelial Caco-2 cells 丁酸钠可预防脂多糖诱导的炎症，恢复人上皮Caco-2细胞紧密连接蛋白的表达。

IF 3.7 4区医学 Q2 CELL BIOLOGY

Cellular immunology

Pub Date : 2025-02-01 DOI: 10.1016/j.cellimm.2024.104912

Jyotsana Bakshi, K.P. Mishra

The gastrointestinal (GI) tract is susceptible to damage under high altitude hypoxic conditions, leading to gastrointestinal discomfort and intestinal barrier injury. Sodium butyrate, a short-chain fatty acid present as a metabolite in the gut, has emerged as a promising therapeutic agent due to its ability to act as an immunomodulatory agent and restore intestinal barrier integrity. This study aimed to explore the mechanism by which sodium butyrate exhibits anti inflammatory effect on intestinal epithelial cells. In vitro, Caco-2 epithelial cells and RAW 264.7 macrophages were used to investigate the protective role of sodium butyrate on Lipopolysaccharide (LPS) induced inflammation. Cell viability assays demonstrated that 1 mM (110.86 μg/mL) of sodium butyrate did not exhibit cytotoxicity on cells in vitro. Treatment with sodium butyrate suppressed reactive oxygen species levels and TNF-α production in LPS-stimulated macrophages, indicating its efficacy in mitigating inflammatory responses. Western blot analysis revealed that sodium butyrate attenuated the expression of iNOS in RAW 264.7 macrophage cells. Moreover, sodium butyrate also reversed the LPS induced over expression of HIF-1α, NLRP3, IL-1β as well as NF-kB in Caco-2 epithelial cells and also had a suppressive effect on IL-8 secretion after LPS stimulation. Immunocytochemistry demonstrated that sodium butyrate enhanced tight junction protein occludin expression in Caco-2 cells while also restoring the decreased permeability of the Caco-2 monolayer due to LPS. These results indicate that sodium butyrate may influence immune responses by suppressing inflammatory mediators and improving the integrity of the epithelial barrier. Understanding the intricate interactions between gut metabolites and host immune responses may help in the development of innovative therapeutic strategies to alleviate intestinal inflammation in high altitude environments.

在高海拔缺氧条件下，胃肠道容易受到损伤，导致胃肠道不适和肠屏障损伤。丁酸钠是一种短链脂肪酸，作为肠道代谢物存在，由于其作为免疫调节剂和恢复肠道屏障完整性的能力，已成为一种有前景的治疗剂。本研究旨在探讨丁酸钠对肠上皮细胞抗炎作用的机制。体外实验采用Caco-2上皮细胞和RAW 264.7巨噬细胞研究丁酸钠对脂多糖（LPS）诱导炎症的保护作用。细胞活力实验表明，1 mM （110.86 μg/mL）丁酸钠对体外培养的细胞无杀伤作用。用丁酸钠治疗可抑制lps刺激的巨噬细胞中活性氧水平和TNF-α的产生，表明其减轻炎症反应的功效。Western blot分析显示，丁酸钠能降低巨噬细胞中iNOS的表达。此外，丁酸钠还能逆转LPS诱导的Caco-2上皮细胞中HIF-1α、NLRP3、IL-1β和NF-kB的过表达，并抑制LPS刺激后IL-8的分泌。免疫细胞化学表明，丁酸钠增强Caco-2细胞中紧密连接蛋白occludin的表达，同时也恢复了由于LPS导致的Caco-2单层通透性下降。这些结果表明，丁酸钠可能通过抑制炎症介质和改善上皮屏障的完整性来影响免疫反应。了解肠道代谢物与宿主免疫反应之间复杂的相互作用可能有助于开发创新的治疗策略，以减轻高海拔环境下的肠道炎症。

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引用次数: 0

Berberine shaping the tumor immune landscape via pyroptosis 小檗碱通过焦亡形成肿瘤免疫景观。

IF 3.7 4区医学 Q2 CELL BIOLOGY

Cellular immunology

Pub Date : 2025-02-01 DOI: 10.1016/j.cellimm.2024.104908

Jinjin Xie , Xin Du , Yuke Li , Chengyu Wu , Rui Li , Mengnan Zhao , Sanjun Shi

Pyroptosis is a programmed cell death (PCD) mainly mediated by the Gasdermin family of proteins, among which Gasdermin E (GSDME) is considered a tumor suppressor gene. GSDME can recruit immune cells to the tumor microenvironment (TME) and promote their effects. Activating and enhancing adaptive immunity through GSDME is a potential solution for anti-tumor therapy. Here we reported that berberine (BBR), a small molecule from traditional Chinese medicine, as a GSDME activator, induced caspase-3 (C-3)/GSDME pathway-mediated pyroptosis through the mitochondrial pathway, improved the immunosuppressive state of the tumor microenvironment, and thus promoted anti-tumor immunity. We determined the induction of pyroptosis of 4 T1 cells by BBR through various experiments, and investigated the immune activation effect of BBR by co-culture in vitro, which induced DCs maturation and macrophage polarization. Zebrafish embryo toxicity experiments were used to evaluate the in vivo safety of berberine. Furthermore, the in vivo antitumor and immune activation effects of BBR were investigated using 4 T1 orthotopic model mice, and the results showed that BBR could eliminate orthotopic tumor cells by activating local and systemic immunity. Moreover, we observed that BBR significantly inhibited breast cancer lung metastasis. In summary, our results showd the role of BBR as a GSDME activator stimulated both local and systemic antitumor immune responses by inducing pyroptosis, effectively preventing tumor development and metastasis.

焦亡是一种主要由Gasdermin家族蛋白介导的程序性细胞死亡（PCD），其中Gasdermin E （GSDME）被认为是一种肿瘤抑制基因。GSDME可以将免疫细胞招募到肿瘤微环境（tumor microenvironment， TME）并促进其作用。通过GSDME激活和增强适应性免疫是抗肿瘤治疗的潜在解决方案。本文报道了中药小分子小檗碱（berberine， BBR）作为GSDME激活剂，通过线粒体途径诱导caspase-3 (C-3)/GSDME途径介导的焦亡，改善肿瘤微环境的免疫抑制状态，从而促进抗肿瘤免疫。我们通过各种实验确定BBR对4个T1细胞的诱导凋亡作用，并通过体外共培养研究BBR诱导DCs成熟和巨噬细胞极化的免疫激活作用。采用斑马鱼胚胎毒性实验评价小檗碱的体内安全性。利用4只T1原位模型小鼠研究了BBR的体内抗肿瘤和免疫激活作用，结果表明BBR可以通过激活局部和全身免疫来消除原位肿瘤细胞。此外，我们观察到BBR显著抑制乳腺癌肺转移。综上所述，我们的研究结果表明，BBR作为GSDME激活剂通过诱导焦亡刺激局部和全身的抗肿瘤免疫反应，有效地阻止肿瘤的发展和转移。

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引用次数: 0

Bacteriophage derived dsRNA induces polarized activation of alveolar macrophages from Balb/c and C57Bl/6 mice in vitro in sex- and age-dependent manner 噬菌体来源的dsRNA诱导Balb/c和C57Bl/6小鼠肺泡巨噬细胞在体外以性别和年龄依赖的方式极化活化。

IF 3.7 4区医学 Q2 CELL BIOLOGY

Cellular immunology

Pub Date : 2025-02-01 DOI: 10.1016/j.cellimm.2025.104916

R. Dovhyi , A. Dvukhriadkina , K. Ostrovska , M. Rudyk , Irina Verhovcova , Kristine Vaivode , D. Pjanova , L. Ostapchenko , L. Skivka

Bacteriophage-derived dsRNA (bp-dsRNA), also known as Larifan, is a poly-functional and wide-spectrum antiviral medication with potent interferonogenic activity. In the lungs of golden Syrian hamsters infected with SARS-CoV-2, Larifan substantially reduces viral load and decreases infection-induced pathological lesion severity. Alveolar macrophages (AM) are key sentinel cells in the lung, which play an important role in antiviral innate immune responses and, at the same time, can trigger infection-associated hyper-inflammatory response. This study revealed that treatment with bp-dsRNA (Larifan) in vitro modulates the functional profile of AM from intact Balb/c and C57Bl/6 mice. The pattern of the drug response depends on the animal strain, age and sex. AM from Balb/c mice generated a weaker response to the preparation as compared to cells from C57Bl/6 mice. Most emphatic responses to the treatment with bf-dsRNA (Larifan) were registered in AM from old males of both BALB/c and C57BL/6 strains with the strongest in the latter. AM from old C57BL/6 females were less likely to be influenced by the preparation. In most cases, exposure to bf-dsRNA (Larifan) increased AM phagocytic activity and was more often accompanied by the stimulation of intracellular reactive oxygen species generation, than by its decrease. In most animal groups, treatment with bf-dsRNA (Larifan) did not affect significantly CD206 expression and down-regulated CD80 expression in AM. Taken together, our findings suggest that bf-dsRNA (Larifan) not so much stimulates the bivalent phenotype of AM, as restrains their hyper-inflammatory responses through the control of antigen-presentation while preserving functional signatures typical of patrolling tissue-resident macrophages.

噬菌体衍生的dsRNA (bp-dsRNA)，也被称为Larifan，是一种多功能广谱抗病毒药物，具有强干扰素活性。在感染SARS-CoV-2的金色叙利亚仓鼠肺中，拉里凡显著降低病毒载量，降低感染引起的病理病变严重程度。肺泡巨噬细胞（Alveolar macrophages， AM）是肺部关键的前哨细胞，在抗病毒先天免疫应答中发挥重要作用，同时可引发感染相关的超炎症反应。本研究表明，bp-dsRNA （Larifan）在体外可调节完整Balb/c和C57Bl/6小鼠AM的功能谱。药物反应的模式取决于动物的品系、年龄和性别。与来自C57Bl/6小鼠的细胞相比，来自Balb/c小鼠的AM对该制剂的反应较弱。BALB/c和C57BL/6菌株的老年男性AM对bf-dsRNA （Larifan）治疗的反应最强烈，后者最强。老年C57BL/6雌性的AM受制剂的影响较小。在大多数情况下，暴露于bf-dsRNA （Larifan）会增加AM的吞噬活性，并且通常伴随着细胞内活性氧生成的刺激，而不是其减少。在大多数动物组中，用bf-dsRNA （Larifan）治疗AM不显著影响CD206表达，并下调CD80表达。综上所述，我们的研究结果表明，bf-dsRNA （Larifan）并不会刺激AM的二价表型，而是通过控制抗原呈递来抑制AM的超炎症反应，同时保留巡逻组织内巨噬细胞的典型功能特征。

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引用次数: 0

Enhanced apoptosis and inflammation allied with autophagic and apoptotic Leishmania amastigotes in the seemingly undamaged ear skin of clinically affected dogs with canine visceral Leishmaniasis 在临床感染犬内脏利什曼病的犬耳皮肤中，与自噬和凋亡利什曼原虫无毛线虫相关的细胞凋亡和炎症增强。

IF 3.7 4区医学 Q2 CELL BIOLOGY

Cellular immunology

Pub Date : 2025-02-01 DOI: 10.1016/j.cellimm.2024.104909

Barbara Laurice Araújo Verçosa , Maria Imaculada Muniz-Junqueira , Ana Lys Bezerra Barradas Mineiro , Maria Norma Melo , Anilton Cesar Vasconcelos

Programmed cell death plays a relevant role in the pathogenesis of visceral Leishmaniasis. Apoptosis selects suitable parasites, regulating parasite density, whereas autophagy eliminates pathogens. This study aimed to assess the inflammation and apoptosis in inflammatory cells and presents a unique description of the presence of autophagic and apoptotic Leishmania amastigotes in naturally Leishmania-infected dogs. Fragments from seemingly undamaged ear skin of sixteen Leishmania-infected dogs and seven uninfected dogs were evaluated through histomorphometry, ultrastructural, immunohistochemical and transmission electron microscopy (TEM) analyses. Leishmania amastigotes were present on seemingly undamaged ear skin only in clinically affected dogs. Parasite load, morphometrical parameters of inflammation and apoptotic index of inflammatory cells were higher in clinically affected animals and were related to clinical manifestations. Apoptotic index and morphometric parameters of the inflammatory infiltrate in undamaged ear skin were positively correlated with parasite load. Apoptotic and non-apoptotic Leishmania amastigotes were observed within neutrophils and macrophages. Leishmania amastigotes were positive for Bax, a marker for apoptosis, by immunohistochemistry. Morphological characteristics of apoptosis and autophagy in Leishmania amastigotes were observed only in phagocytes of clinically affected dogs. Positive correlations were found between histomorphometry and clinical manifestations. Our results showed that apoptosis and autophagy in Leishmania amastigotes may be related to both the increase in parasite load and apoptotic index in inflammatory cells, and with the intensity of the inflammatory response in clinically affected dogs. Thus, our study suggests that apoptotic and autophagy Leishmania within phagocytes may have facilitate the survival of the parasite and it appears to play an important role in the process of Leishmania infection.

程序性细胞死亡在内脏利什曼病的发病机制中起着相关作用。细胞凋亡选择合适的寄生虫，调节寄生虫密度，而自噬消除病原体。本研究旨在评估炎症细胞的炎症和凋亡，并对自然感染利什曼原虫的狗体内存在自噬和凋亡的利什曼原虫无鞭毛体进行独特的描述。对16只感染利什曼犬和7只未感染利什曼犬的耳部皮肤进行组织形态学、超微结构、免疫组织化学和透射电镜分析。利什曼原虫无鞭毛体只存在于临床感染犬的看似未受损的耳皮肤上。临床感染动物的寄生虫负荷、炎症形态计量参数和炎症细胞凋亡指数均较高，且与临床表现有关。未损伤耳部皮肤炎症浸润的凋亡指数和形态学参数与寄生虫负荷呈正相关。中性粒细胞和巨噬细胞内均可见凋亡和非凋亡利什曼原虫。免疫组化法检测利什曼原虫凋亡标志物Bax阳性。利什曼原虫凋亡和自噬的形态学特征仅在临床感染犬的吞噬细胞中观察到。组织形态学与临床表现呈正相关。我们的研究结果表明，利什曼原虫的凋亡和自噬可能与炎症细胞中寄生虫负荷和凋亡指数的增加有关，并与临床感染犬的炎症反应强度有关。因此，我们的研究表明，吞噬细胞内的凋亡和自噬利什曼原虫可能促进了寄生虫的生存，并在利什曼原虫感染过程中发挥了重要作用。

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引用次数: 0

P2RX1-blocked neutrophils induce CD8+ T cell dysfunction and affect the immune escape of gastric cancer cells p2rx1阻断的中性粒细胞诱导CD8+ T细胞功能障碍，影响胃癌细胞的免疫逃逸。

IF 3.7 4区医学 Q2 CELL BIOLOGY

Cellular immunology

Pub Date : 2025-02-01 DOI: 10.1016/j.cellimm.2024.104901

Yan Zhang , Fenglin Zhang , Zhi Liu , Min Li , Ge Wu , Hui Li

Background

Gastric cancer (GC) is one of the deadly malignancies of the gastrointestinal tract. Research has confirmed the linkage of P2RX1 with immune cell activation and tumor progression. This project focused on the impact of P2RX1 level in neutrophils on the efficacy of immune checkpoint inhibitor (ICI) treatment in GC.

Methods

Blood samples from 23 GC patients eligible for camrelizumab treatment were collected. Flow cytometry was carried out to analyze the proportion of P2RX1 in neutrophils. IHC was utilized to detect the expression level of PD-L1. We also evaluated the chemotaxis ability of neutrophils using a Transwell system, assessed the viability and apoptosis rate of GC cells using CCK-8 and flow cytometry, measured the proportions of CD8⁺PD-1⁺ and CD8⁺GZMB⁺ cells, determined the expression levels of IL-6, TNFα, IFN-γ, IL-8, IL-12, IL-1β, and GZMB by utilizing enzyme-linked immunosorbent assay (ELISA), and examined the expression levels of P2RX1 and PD-L1 using western blot (WB). By establishing a xenograft mouse model, we studied the impact of P2RX1-blocked neutrophils on the efficacy of ICI treatment in the GC microenvironment.

Results

In GC, clinical analysis revealed increased infiltration of P2RX1-lowly expressed neutrophil subsets and increased expression of PD-L1. In vitro experiments demonstrated that abnormal expression of P2RX1 affected neutrophil function. Furthermore, the blockage or knockdown of P2RX1 in neutrophils modulated CD8⁺ T cell function, promoting GC progression. In in vivo experiments, the blockage of P2RX1 in neutrophils inhibited the effectiveness of ICI treatment in the GC microenvironment.

Conclusion

This project validated that the loss of P2RX1 in neutrophils induces CD8⁺ T cell dysfunction and affects the GC development, indicating that P2RX1 may be an accurate biomarker for predicting ICI response, thus providing a theoretical basis for the clinical application of ICI.

背景：胃癌（GC）是胃肠道中致命的恶性肿瘤之一。研究证实 P2RX1 与免疫细胞活化和肿瘤进展有关。本项目重点研究中性粒细胞中P2RX1水平对免疫检查点抑制剂（ICI）治疗胃癌疗效的影响：方法：收集了23名符合康瑞珠单抗治疗条件的GC患者的血液样本。流式细胞术分析了中性粒细胞中 P2RX1 的比例。利用 IHC 检测 PD-L1 的表达水平。我们还使用 Transwell 系统评估了中性粒细胞的趋化能力，使用 CCK-8 和流式细胞术评估了 GC 细胞的存活率和凋亡率，测量了 CD8+PD-1+ 和 CD8+GZMB+ 细胞的比例、利用酶联免疫吸附试验（ELISA）测定 IL-6、TNFα、IFN-γ、IL-8、IL-12、IL-1β 和 GZMB 的表达水平，并利用 Western 印迹（WB）检测 P2RX1 和 PD-L1 的表达水平。通过建立异种移植小鼠模型，我们研究了 P2RX1 受体阻断的中性粒细胞对 ICI 治疗 GC 微环境疗效的影响：在 GC 中，临床分析显示 P2RX1 低表达的中性粒细胞亚群浸润增加，PD-L1 表达增加。体外实验表明，P2RX1 的异常表达会影响中性粒细胞的功能。此外，阻断或敲除中性粒细胞中的 P2RX1 会调节 CD8+ T 细胞的功能，促进 GC 的进展。在体内实验中，阻断中性粒细胞中的 P2RX1 会抑制 ICI 在 GC 微环境中的治疗效果：该项目验证了中性粒细胞中P2RX1的缺失会诱导CD8+ T细胞功能障碍并影响GC的发展，表明P2RX1可能是预测ICI反应的准确生物标志物，从而为ICI的临床应用提供了理论依据。

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引用次数: 0