Developmental and comparative immunology最新文献_第4页

Characterization of the CD28 signaling pathway related genes (CD28, Grb2 and SLP-76) of golden pompano (Trachinotus ovatus) 金鲳鱼CD28信号通路相关基因（CD28、Grb2和SLP-76）的研究

IF 2.4 3区农林科学 Q1 FISHERIES

Developmental and comparative immunology

Pub Date : 2025-12-01 DOI: 10.1016/j.dci.2025.105529

Zhitao Qi , Jinquan He , Chenfei Gu , Yushuai Xie , Mingzhu Pan , Qihuan Zhang , Youchuan Wei

CD28 signaling pathway plays critical roles in the T cell activation. The golden pompano (Trachinotus ovatus, Tro) is a commercially important marine fish that widely cultured in southern China. Frequently pathogenic disease outbreaks greatly hinder the industry of golden pompano. Understanding the T cell activation signaling pathway of golden pompano contributes to prevent pathogenic diseases using immunological strategies. In the present study, three molecules of CD28 signaling pathway (TroCD28, growth factor receptor-bound protein 2 (TroGrb2) and SH2 domain-containing leukocyte phosphoprotein of 76 kDa (TroSLP-76)) were identified from golden pompano. The cDNA of the three genes respectively encoded 248, 217 and 375 amino acids. TroCD28, TroGrb2 and TroSLP-76 contained several conserved functional domains, similar to their mammalian counterparts. qPCR analysis revealed that these three genes were ubiquitously expressed in all selected tissues and peripheral blood lymphocytes (PBLs), with high expressions in immune-related tissues (spleen, head kidney, gill). Further, expressions of these three genes in head kidney were significantly induced at different time points post lipopolysaccharide (LPS) or polyriboinosinic polyribocytidylic acid (polyI:C) stimulation and Vibrio alginolyticus infection. The coimmunoprecipitation (Co-IP) assay verified the TroCD28-TroGrb2 interaction. These results provide basis for understanding the process of T cell activation in golden pompano.

CD28信号通路在T细胞活化中起关键作用。金鲳鱼（Trachinotus ovatus, Tro）是中国南方广泛养殖的一种重要的商业海鱼。病原性疾病的频繁爆发严重阻碍了金鲳鱼产业的发展。了解鲳鱼的T细胞激活信号通路有助于利用免疫策略预防致病性疾病。本研究从金鲳鱼中鉴定出3个CD28信号通路分子（TroCD28、生长因子受体结合蛋白2 （TroGrb2）和含SH2结构域的白细胞磷酸化蛋白76 kDa (TroSLP-76)）。三个基因的cDNA分别编码248个、217个和375个氨基酸。TroCD28、TroGrb2和TroSLP-76含有几个保守的功能域，与哺乳动物相似。qPCR分析显示，这三个基因在所有选择的组织和外周血淋巴细胞（PBLs）中普遍表达，在免疫相关组织（脾、头肾、鳃）中表达量高。此外，这三个基因在脂多糖（LPS）或多核糖素多核糖素酸（polyI:C）刺激和溶藻弧菌感染后的不同时间点显著诱导头肾的表达。共免疫沉淀（Co-IP）实验证实了TroCD28-TroGrb2相互作用。这些结果为了解金鲳鱼体内T细胞的活化过程提供了依据。

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引用次数: 0

Transcription factor expression for identifying B cell subsets in cattle by flow cytometry 流式细胞术鉴定牛B细胞亚群的转录因子表达

IF 2.4 3区农林科学 Q1 FISHERIES

Developmental and comparative immunology

Pub Date : 2025-12-01 DOI: 10.1016/j.dci.2025.105531

Hayley Brown , Selma Schmidt , Michelle Thom , Adam Kuča , Claudine Porta , John A. Hammond , Marie Bonnet-Di Placido , Wilhelm Gerner

With their capacity for antibody production, B cells underpin many protective immune responses, but they can also contribute to immunopathology and autoimmunity. Studying their differentiation following antigen encounter is therefore a key aspect of fundamental immunological research. However, in many livestock species, including cattle, the study of B cell subsets is limited by the restricted availability of immunological tools. Here, we demonstrate that cross-reacting anti-human and anti-murine antibodies used in combination with established anti-bovine reagents can be used to identify three major cattle B cell subsets by flow cytometry: a) total mature B cells, b) plasma cells and plasmablasts, and c) germinal centre (GC) B cells. For this, we employed antibodies targeting CD79α, Pax5, Blimp1, IRF4, and Bcl6. CD79α/Pax5 co-expressing total B cells largely overlapped with the previously described CD14⁻CD40⁺ phenotype. Putative plasma cells, defined by Blimp1/IRF4 co-expression, were detected in blood and lymph nodes and exhibited reduced Pax5 expression, consistent with a plasma cell phenotype. In contrast, putative GC B cells, identified by Bcl6 expression, were only observed in lymph nodes, as expected for this population. Using foot-and-mouth disease virus-like particles (VLPs) as probes to detect antigen-specific B cells in blood and lymph nodes from VLP-immunised cattle, we show that our newly established phenotypes facilitate the dissection of antigen-driven B cell responses. Notably, VLP-binding B cell subsets were predominantly detected in lymph node cell preparations draining the site of vaccination. Thus, this work significantly expands the toolbox available for studying B cell biology in cattle.

B细胞具有产生抗体的能力，支持许多保护性免疫反应，但它们也可以促进免疫病理和自身免疫。因此，研究它们在抗原接触后的分化是基础免疫学研究的一个关键方面。然而，在包括牛在内的许多家畜物种中，B细胞亚群的研究受到免疫工具有限可用性的限制。在这里，我们证明了交叉反应的抗人和抗鼠抗体与已建立的抗牛试剂联合使用，可以通过流式细胞术识别三种主要的牛B细胞亚群：a)总成熟B细胞，B)浆细胞和浆母细胞，以及c)生发中心（GC） B细胞。为此，我们采用了靶向CD79α、Pax5、Blimp1、IRF4和Bcl6的抗体。共表达CD79α/Pax5的总B细胞与先前描述的CD14−CD40+表型在很大程度上重叠。在血液和淋巴结中检测到由Blimp1/IRF4共表达定义的推定浆细胞，Pax5表达减少，与浆细胞表型一致。相比之下，通过Bcl6表达鉴定的GC B细胞仅在淋巴结中观察到，这与该人群的预期一致。使用口蹄疫病毒样颗粒（vlp）作为探针检测vlp免疫牛的血液和淋巴结中的抗原特异性B细胞，我们发现我们新建立的表型有助于抗原驱动B细胞反应的解剖。值得注意的是，vlp结合的B细胞亚群主要在疫苗接种部位的淋巴结细胞制剂中检测到。因此，这项工作大大扩展了研究牛B细胞生物学的工具箱。

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引用次数: 0

Exosomal miRNA-mediated enhancement of hemocyte immunity in Vibrio-stressed blood clam (Anadara granosa) 外泌体mirna介导的应激弧菌血蛤血细胞免疫增强。

IF 2.4 3区农林科学 Q1 FISHERIES

Developmental and comparative immunology

Pub Date : 2025-11-30 DOI: 10.1016/j.dci.2025.105533

Shanjie Zha , Siyi Huang , Luxia Zhang , Xinguo Zhao , Hongxing Liu , Wei Shi , Yongbo Bao

Exosomes are crucial for intercellular immune communication, but their function in marine bivalve immunity remains poorly understood. This study investigated the immunomodulatory role of hemocyte-derived exosomes in the blood clam Anadara granosa during Vibrio parahaemolyticus challenge. Exosomes isolated from hemolymph displayed typical cup-shaped morphology (149–153 nm diameter) and expressed canonical markers (CD63, TSG101). Functional analysis demonstrated that exosomes from Vibrio-challenged clams significantly enhanced hemocyte phagocytosis and reduced bacterial loads. To uncover the underlying mechanism, small RNA sequencing of the exosomal cargo identified 107 differentially expressed miRNAs (56 upregulated, 51 downregulated) upon bacterial challenge. Bioinformatic analysis revealed that these miRNAs primarily target genes involved in endocytosis, autophagy, and ubiquitin-mediated proteolysis. Notably, quantitative PCR validation confirmed the differential expression of six key miRNAs involved in phagocytosis regulation. These findings provide the first evidence that A. granosa hemocytes utilize exosomal miRNAs to regulate antibacterial response by enhanced phagocytosis. The identified exosomal miRNAs represent promising biomarkers for immune assessment and potential targets for developing RNA-based therapeutics to enhance disease resistance aquaculture.

外泌体对细胞间免疫通讯至关重要，但它们在海洋双壳类动物免疫中的功能尚不清楚。本研究探讨了血细胞来源的外泌体在副溶血性弧菌侵袭下对血蛤的免疫调节作用。从血淋巴中分离的外泌体呈现典型的杯状形态（直径149 ~ 153 nm），并表达典型标记（CD63、TSG101）。功能分析表明，来自弧菌攻击蛤的外泌体显着增强血细胞吞噬并减少细菌负荷。为了揭示潜在的机制，外泌体货物的小RNA测序鉴定了107个差异表达的mirna（56个上调，51个下调）。生物信息学分析显示，这些mirna主要针对参与内吞作用、自噬和泛素介导的蛋白质水解的基因。值得注意的是，定量PCR验证证实了参与吞噬调节的六个关键mirna的差异表达。这些发现提供了第一个证据，表明大粒棘球蚴血细胞利用外泌体mirna通过增强吞噬来协调强大的抗菌反应。鉴定的外泌体mirna代表了有希望的免疫评估生物标志物和开发基于rna的治疗方法以增强水产养殖抗病能力的潜在靶点。

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引用次数: 0

Genome-wide analysis unveils the differentiated “external” and “internal” lectin system in the tropical sea cucumber Holothuria leucospilota 全基因组分析揭示了热带海参Holothuria leucospilota分化的“外部”和“内部”凝集素系统

IF 2.4 3区农林科学 Q1 FISHERIES

Developmental and comparative immunology

Pub Date : 2025-11-20 DOI: 10.1016/j.dci.2025.105527

Xinyue Ren , Zhou Qin , Renhui Liu , Ting Chen , Tiehao Lin , Xinyu Sun , Jiasheng Huang , Wenjie Pan , Caixia Long , Chunhua Ren , Haipeng Qin , Peiming Zheng , Aifen Yan , Lihong Yuan

To date, the understanding of lectin-related immunity in sea cucumbers remains limited. Through a comprehensive genome-wide analysis, 133 lectin genes were identified from Holothuria leucospilota, a sea cucumber species with high ecological and economic significance in the Pacific and Indian Oceans. These lectins were classified into five subtypes, with C-type lectin being the predominant form, and the presence of SUEL-like lectin and Malectin reported here for the first time in sea cucumbers. Transcriptomic analysis across twelve tissues and sixteen developmental stages revealed ubiquitous expression of H. leucospilota lectins. A key finding was the compartmentalization of lectin systems: SUEL-like lectins expressed in the body wall constitute an “external” defense system, while C-type lectins produced by coelomocytes form an “internal” immune system. This spatial and functional specialization reflects a sophisticated immune strategy in invertebrates, enabling targeted pathogen recognition at different bodily interfaces. Furthermore, distinct immune responses to Vibrio, LPS, and poly(I:C) were observed among different lectins. We also identified an ovary-specific C-type lectin, suggesting a potential role in early embryonic development. This study not only fills a knowledge gap regarding lectin genes in sea cucumbers but also establishes a broader framework for understanding complex immune mechanisms in echinoderms.

迄今为止，对海参凝集素相关免疫的认识仍然有限。通过全面的全基因组分析，从太平洋和印度洋具有高度生态和经济意义的海参物种leucospilota中鉴定出133个凝集素基因。这些凝集素可分为5个亚型，以c型凝集素为主要亚型，本文首次报道了suel样凝集素和Malectin在海参中的存在。对12个组织和16个发育阶段的转录组学分析显示，蓝褐藻凝集素的表达是普遍存在的。一个关键的发现是凝集素系统的区隔化：在体壁表达的suel样凝集素构成了一个“外部”防御系统，而由体腔细胞产生的c型凝集素形成了一个“内部”免疫系统。这种空间和功能专门化反映了无脊椎动物复杂的免疫策略，能够在不同的身体界面上识别目标病原体。此外，不同的凝集素对弧菌、脂多糖和聚（I:C）的免疫反应也不同。我们还发现了一种卵巢特异性c型凝集素，这表明它在早期胚胎发育中具有潜在作用。本研究不仅填补了海参凝集素基因的知识空白，而且为理解棘皮动物复杂的免疫机制建立了一个更广泛的框架。

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引用次数: 0

Virus-like particles of Macrobrachium rosenbergii nodavirus displaying M2e of influenza A viruses protect White Leghorn chickens against heterologous H9N2 and H5N2 challenges 显示甲型流感病毒M2e的罗氏沼虾诺达病毒样颗粒保护白来窝鸡免受异源H9N2和H5N2的攻击。

IF 2.4 3区农林科学 Q1 FISHERIES

Developmental and comparative immunology

Pub Date : 2025-11-18 DOI: 10.1016/j.dci.2025.105526

Bernard Yi Zhe Thian , Mohd Nasir Nurul Fatimah , Abdul Rahman Omar , Huzlinda Hussin , Hui Kian Ong , Chuan Loo Wong , Abdul Razak Mariatulqabtiah , Kok Lian Ho , Wen Siang Tan

H9N2 and H5N2 avian influenza A viruses (IAVs) are two of the most widespread and economically damaging subtypes affecting the poultry industry. Controlling these viruses in poultry remains challenging, highlighting the need for a vaccine that can confer broad and effective protection across multiple avian IAV subtypes. Previously, we fused two copies of the matrix 2 ectodomain (M2e) of human and avian IAV (2xhuM2e-2xavM2e) at the C-terminus of Macrobrachium rosenbergii nodavirus capsid protein (NvC). This chimeric protein (NvC-2xhuM2e-2xavM2e) self-assembled into virus-like particles (VLPs), which elicited broad immune responses and activated humoral and cellular immune responses in chickens. In this study, we evaluated the protective efficacy of these chimeric VLPs in White Leghorn chickens against H9N2 and H5N2 infections. Chickens immunized with the chimeric VLPs produced a significant level of M2e antibodies that reacted with the M2e of H9N2 and H5N2 IAVs. Following viral infections, the chimeric VLPs mitigated weight loss in H9N2-infected chickens and alleviated delayed weight gain in H5N2-infected chickens. Furthermore, the chimeric VLPs reduced the clinical signs in immunized chickens challenged with either H9N2 or H5N2 avian IAV. Viral loads in the lung tissues of the immunized chickens were significantly lower compared to the negative control group. The chimeric VLPs also enhanced H9N2 and H5N2 viral clearance, showing a significant reduction in viral shedding in the oropharynges and cloacae of the immunized chickens. Furthermore, the VLPs activated both inflammatory and anti-inflammatory cytokines, which were associated with enhanced viral clearance while reducing lung and tracheal immunopathology. Collectively, these findings demonstrate the potential of the chimeric VLPs as a cross-protective vaccine against both H9N2 and H5N2 infections in poultry.

H9N2和H5N2甲型禽流感病毒（iav）是影响家禽业的两种最广泛和最具经济破坏性的亚型。在家禽中控制这些病毒仍然具有挑战性，这突出表明需要一种能够对多种禽流感亚型提供广泛和有效保护的疫苗。在此之前，我们在罗氏螯虾诺达病毒衣壳蛋白（NvC）的c端融合了人类和禽类IAV的两个拷贝（2xhuM2e-2xavM2e）的基质2外域（M2e）。这种嵌合蛋白（NvC-2xhuM2e-2xavM2e）自组装成病毒样颗粒（vlp），在鸡体内引起广泛的免疫反应，并激活体液和细胞免疫反应。在本研究中，我们评估了这些嵌合vlp对白来角鸡对H9N2和H5N2感染的保护作用。嵌合vlp免疫的鸡产生了显著水平的M2e抗体，与H9N2和H5N2 iav的M2e反应。病毒感染后，嵌合vlp减轻了h9n2感染鸡的体重减轻，并减轻了h5n2感染鸡的延迟体重增加。此外，嵌合vlp降低了感染H9N2或H5N2禽流感病毒的免疫鸡的临床症状。免疫组鸡肺组织病毒载量显著低于阴性对照组。嵌合vlp还增强了H9N2和H5N2病毒的清除率，显示免疫鸡口咽和泄殖腔的病毒脱落显著减少。此外，VLPs激活炎症和抗炎细胞因子，这与增强病毒清除有关，同时减少肺和气管的免疫病理。总的来说，这些发现证明了嵌合VLPs作为家禽H9N2和H5N2感染的交叉保护疫苗的潜力。

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引用次数: 0

Acute intestinal injury in hybrid grouper (Epinephelus fuscoguttatus ♀ × Epinephelus lanceolatus ♂) under infection with Vibrio harveyi 杂交石斑鱼（斑石斑鱼♀× lanceolatus Epinephelus♂）感染哈威弧菌后急性肠道损伤的研究

IF 2.4 3区农林科学 Q1 FISHERIES

Developmental and comparative immunology

Pub Date : 2025-11-17 DOI: 10.1016/j.dci.2025.105525

Zhuojin He , Xiaomin Zheng , Lixin Ma, Zhilong Chen, Li Lin, Fei Shi

Vibrio harveyi is a major marine fish pathogen, yet the intestinal pathology and immune defense mechanisms during acute infection remain unclear. The effects of V. harveyi infection on intestinal structure, biochemical indicators, and transcriptomic profiles of hybrid grouper (Epinephelus fuscoguttatus ♀ × Epinephelus lanceolatus ♂) were investigated. The results showed that V. harveyi infection significantly increased goblet cell hyperplasia (P < 0.05), thinning of the midgut wall (P < 0.05), and intestinal apoptosis (P < 0.05). Biochemical assays revealed significantly up-regulated activities of catalase (CAT) (P < 0.05) and superoxide dismutase (SOD) (P < 0.05), accompanied by decreased glutathione (GSH) levels (P < 0.05) and increased malondialdehyde (MDA) content (P < 0.01). Transcriptomic analysis revealed differentially expressed genes were enriched in the Antigen processing and presentation pathway and Cytokine-cytokine receptor interaction pathway. Additionally, quantitative real-time PCR demonstrated increased mRNA expression of Interleukin-6 (IL-6), Nuclear Factor Kappa-B p65 (NF-κB p65), and the tight-junction protein Claudin-3α (CLDN-3α). Redundancy analysis identified significant correlations among oxidative stress indicators, differentially expressed genes, and intestinal morphometric parameters (goblet cell number, villus length and intestinal wall thickness). Collectively, these findings highlight that V. harveyi compromises hybrid grouper health through disruption of mucosal structure, oxidative homeostasis, and immune regulation.

哈氏弧菌是一种主要的海鱼致病菌，但其急性感染的肠道病理和免疫防御机制尚不清楚。研究了harveyi弧菌感染对杂交石斑鱼（斑石斑鱼♀× lanceolatus Epinephelus fuscoguttatus♀）肠道结构、生化指标和转录组学的影响。结果表明，感染V. harveyi显著增加了杯状细胞增生（P < 0.05）、中肠壁变薄（P < 0.05）和肠道细胞凋亡（P < 0.05）。生化检测结果显示，过氧化氢酶（CAT）和超氧化物歧化酶（SOD）活性显著升高（P < 0.05），谷胱甘肽（GSH）水平显著降低（P < 0.05），丙二醛（MDA）含量显著升高（P < 0.01）。转录组学分析显示，抗原加工递呈途径和细胞因子-细胞因子受体相互作用途径中富集了差异表达基因。此外，实时荧光定量PCR显示白细胞介素-6 （IL-6）、核因子κ b p65 （NF-κB p65）和紧密连接蛋白CLDN-3α的mRNA表达增加。冗余分析发现氧化应激指标、差异表达基因和肠道形态计量参数（杯状细胞数量、绒毛长度和肠壁厚度）之间存在显著相关性。总的来说，这些发现强调了V. harveyi通过破坏粘膜结构、氧化稳态和免疫调节来损害杂交石斑鱼的健康。

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引用次数: 0

Soybean glycinin and β-conglycinin disrupted mitochondria-endoplasmic reticulum interactions in porcine intestinal epithelial cells via oxidative stress cascades 大豆甘氨酸和β-甘氨酸通过氧化应激级联破坏了猪肠上皮细胞线粒体和内质网之间的相互作用。

IF 2.4 3区农林科学 Q1 FISHERIES

Developmental and comparative immunology

Pub Date : 2025-11-15 DOI: 10.1016/j.dci.2025.105523

Lei Wang , Chenlong Wang , Hongyan Ding , Shibin Feng , Xichun Wang , Jinjie Wu , Dong Liu

Soybean glycinin (11S) and β-conglycinin (7S) are major contributors to allergic diarrhea and intestinal barrier damage in young animals. This study investigated the molecular mechanisms underlying the 7S- and 11S-induced dysfunction of mitochondrial and endoplasmic reticulum (ER) interactions in porcine intestinal epithelial (IPEC-J2) cells via the oxidative stress pathway. The results showed that 7S- and 11S-induced oxidative stress, as evidenced by the following findings: reduced manganese superoxide dismutase (Mn-SOD) activity and elevated 8-Hydroxy-2′-deoxyguanosine (8-OHdG) levels, with excessive reactive oxygen species (ROS) accumulation and elevated Ca²⁺ levels; decreased mitochondrial membrane potential (MMP), damaged mitochondria-associated endoplasmic reticulum membranes (MAM) structure; up-regulated the protein expression of glucose-regulated protein 75 (GRP75) and mitochondrial Rho-GTPase 1 (Miro1), while inositol 1,4,5 -trisphosphate receptor (IP3R), voltage-dependent anion channel 1 (VDAC1), mitofusin2 (MFN2) and phosphofurin acidic cluster sorting protein 2 (PACS2) were down-regulated. N-acetylcysteine (NAC)pre-treatment alleviated ROS accumulation and mitigated Ca²⁺ overload and MAM dysfunction, thereby ameliorating IPEC-J2 cell injury. In conclusion, 7S- and 11S-induced ROS burst to disrupt mitochondria-ER interaction homeostasis, leading to MAM structural damage and calcium dysregulation in IPEC-J2 cells, NAC effectively mitigated this process by scavenging ROS. These findings elucidate the critical involvement of subcellular organelle interaction disorders in food allergy pathogenesis and provide novel insights for targeted intervention strategies.

大豆甘氨酸（11S）和β-甘氨酸（7S）是幼龄动物过敏性腹泻和肠屏障损伤的主要原因。本研究通过氧化应激途径研究了7S和11s诱导的猪肠上皮细胞（IPEC-J2）线粒体和内质网（ER）相互作用功能障碍的分子机制。结果表明，7S-和11s诱导的氧化应激，表现为锰超氧化物歧化酶（Mn-SOD）活性降低，8-羟基-2′-脱氧鸟苷（8-OHdG）水平升高，活性氧（ROS）积累过多，Ca2+水平升高；线粒体膜电位降低，线粒体相关内质网膜（MAM）结构受损；上调葡萄糖调节蛋白75 （GRP75）和线粒体Rho-GTPase 1 （Miro1）的蛋白表达，下调肌醇1,4,5 -三磷酸受体（IP3R）、电压依赖性阴离子通道1 （VDAC1）、丝裂酶2 （MFN2）和磷酸氟酸簇分选蛋白2 （PACS2）的蛋白表达。n -乙酰半胱氨酸（NAC）预处理可减轻ROS积累，减轻Ca2+超载和MAM功能障碍，从而改善IPEC-J2细胞损伤。综上所述，7S-和11s诱导的ROS爆发破坏线粒体-内质网相互作用的稳态，导致IPEC-J2细胞MAM结构损伤和钙失调，NAC通过清除ROS有效缓解了这一过程。这些发现阐明了亚细胞细胞器相互作用障碍在食物过敏发病机制中的关键作用，并为有针对性的干预策略提供了新的见解。

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引用次数: 0

NKp46 identifies a subset of goat leukocytes with phenotypic and functional characteristics of natural killer cells NKp46鉴定出山羊白细胞亚群具有自然杀伤细胞的表型和功能特征。

IF 2.4 3区农林科学 Q1 FISHERIES

Developmental and comparative immunology

Pub Date : 2025-11-15 DOI: 10.1016/j.dci.2025.105524

Mahmoud M. Elnaggar , Amany Hassan , Gaber S. Abdellrazeq , Asmaa H. Mahmoud , William C. Davis

Natural killer (NK) cells are the first discovered member of innate lymphoid cells (ILCs) that play a pivotal role in host defense. They were initially identified by their ability to nonspecifically lyse certain tumor and virally infected cells. What is known about NK cells in species other than humans and mice is limited. The progress that has been made is through development and use of monoclonal antibodies (mAbs) specific to NKp46 (NCR1, CD335), a major NK cell-activating receptor that has been used as a marker to identify leukocytes with NK cell characteristics in cattle, sheep, buffaloes, pigs, horses and dogs. As described herein, the finding that the mAb that recognizes sheep and cattle NKp46 recognizes the orthologue in goats has provided an opportunity to identify a subset of leukocytes with phenotypic and functional characteristics of NK cells in goats, a profile consistent with NK cell features reported in other animal species. The study expands opportunities to conduct comparative studies on NK cells across different species and underscores the potential use of goats as an animal model in investigating NK cell biology in health and disease.

自然杀伤细胞（NK）是先天淋巴样细胞（ILCs）中第一个被发现的成员，在宿主防御中起着关键作用。它们最初是通过非特异性地溶解某些肿瘤和病毒感染细胞的能力来鉴定的。除了人类和老鼠之外，对NK细胞的了解是有限的。目前取得的进展是通过开发和使用NKp46 （NCR1, CD335）的单克隆抗体（mab）， NKp46是一种主要的NK细胞激活受体，已被用作识别牛、羊、水牛、猪、马和狗中具有NK细胞特征的白细胞的标记。如本文所述，发现识别绵羊和牛NKp46的单抗识别山羊的同源物，这为鉴定山羊中具有NK细胞表型和功能特征的白细胞亚群提供了机会，这与其他动物物种中报道的NK细胞特征一致。该研究扩大了对不同物种的NK细胞进行比较研究的机会，并强调了山羊作为研究健康和疾病中NK细胞生物学的动物模型的潜力。

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引用次数: 0

Functional characterization of Micropterus salmoides galectin-3 (MsGal-3B): a multifunctional immune effector with agglutination and antibacterial activities 小蜈蚣半凝集素-3 (MsGal-3B)：一种具有凝集和抗菌活性的多功能免疫效应物。

IF 2.4 3区农林科学 Q1 FISHERIES

Developmental and comparative immunology

Pub Date : 2025-11-15 DOI: 10.1016/j.dci.2025.105515

Ying Ye , Ce Zhang , Fengyue Guo , Siyi She , Shun Yang , Mengmeng Huang , Hui Fei

Galectins are evolutionarily conserved lectins that play pivotal roles in innate immunity. In this study, we functionally characterized a novel chimera-type galectin-3 homolog from largemouth bass (Micropterus salmoides), designated MsGal-3B. The open reading frame of MsGal-3B encodes a 258-amino acid protein containing a canonical carbohydrate recognition domain (CRD) and an N-terminal non-lectin region, typical of galectin-3 family members. Phylogenetic analysis revealed that MsGal-3B clusters with galectin-3 orthologs from Perciformes, suggesting functional conservation within this order. Recombinant MsGal-3B (rMsGal-3B) exhibited strong affinity for peptidoglycan (PGN) and β-glucan (GLU), and weak interaction with lipopolysaccharide (LPS). Microbial binding assays revealed broad recognition across Gram-positive bacteria (Micrococcus luteus, Staphylococcus aureus), Gram-negative bacteria (Aeromonas veronii, Vibrio parahaemolyticus), and fungi (Saccharomyces cerevisiae, Pichia pastoris). Notably, rMsGal-3B selectively agglutinated Escherichia coli and M. luteus, but not A. veronii or S. cerevisiae, indicating ligand-dependent functional specificity. rMsGal-3B significantly suppressed the proliferation of E. coli, M. luteus, and S. cerevisiae, while showing no inhibitory effect on A. veronii, suggesting a narrow-spectrum antimicrobial profile. Additionally, rMsGal-3B exhibited hemagglutination activity, implying a potential role in host cell recognition or immune modulation. Collectively, these findings demonstrate that MsGal-3B functions as a multifunctional immune effector in M. salmoides, capable of selective microbial recognition, agglutination, and targeted antimicrobial activity. This study provides new insights into the role of teleost galectins in innate immunity and highlights their potential as molecular targets for disease control in aquaculture.

凝集素是进化上保守的凝集素，在先天免疫中起关键作用。在这项研究中，我们对来自大口黑鲈（Micropterus salmoides）的一种新的嵌合型半凝集素-3同源物进行了功能表征，命名为MsGal-3B。MsGal-3B的开放阅读框编码一个含有典型碳水化合物识别域（CRD）和n端非凝集素区域的258个氨基酸的蛋白，这是半乳糖凝集素-3家族成员的典型特征。系统发育分析显示，MsGal-3B簇与来自Perciformes的半乳糖凝集素-3同源物，表明该目的功能守恒。重组MsGal-3B （rMsGal-3B）对肽聚糖（PGN）和β-葡聚糖（GLU）具有较强的亲和力，与脂多糖（LPS）的相互作用较弱。微生物结合试验显示，革兰氏阳性菌（黄体微球菌、金黄色葡萄球菌）、革兰氏阴性菌（维罗氏气单胞菌、副溶血性弧菌）和真菌（酿酒酵母菌、毕赤酵母）具有广泛的识别性。值得注意的是，rMsGal-3B选择性地凝集了大肠杆菌和黄体芽孢杆菌，而不是维罗氏芽孢杆菌或酿酒葡萄球菌，表明了配体依赖性的功能特异性。rMsGal-3B显著抑制大肠杆菌、黄体分枝杆菌和酿酒酵母的增殖，而对韦氏芽孢杆菌无抑制作用，提示其具有窄谱抗菌谱。此外，rMsGal-3B表现出血凝活性，这意味着它在宿主细胞识别或免疫调节中具有潜在作用。综上所述，这些发现表明MsGal-3B在salmoides中作为一种多功能免疫效应物，具有选择性微生物识别、凝集和靶向抗菌活性。该研究为硬骨鱼凝集素在先天免疫中的作用提供了新的见解，并突出了它们作为水产养殖疾病控制分子靶点的潜力。

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引用次数: 0

Structural characterization of T-cell kinases Lck1 and ZAP-70 in rainbow trout (Oncorhynchus mykiss) through homology modeling 虹鳟（Oncorhynchus mykiss） t细胞激酶Lck1和zak -70的同源性模型分析

IF 2.4 3区农林科学 Q1 FISHERIES

Developmental and comparative immunology

Pub Date : 2025-11-14 DOI: 10.1016/j.dci.2025.105520

Francisco Bravo , Camila Kossack , Claudia Galleguillos-Becerra , Antonella Galleguillos-Becerra , Ricardo Fernández , Kevin Maisey

Early T-cell receptor (TCR) signaling is well defined in mammals but remains underexplored in teleosts such as rainbow trout. The proximal tyrosine kinases Lck and ZAP-70 initiate signaling by phosphorylating and decoding CD3 ITAM motifs. In rainbow trout, two sequences have been annotated for each kinase; their functional relevance is unclear.

We combined multiple sequence alignments, homology modeling (SWISS-MODEL), and structure quality checks (VERIFY3D, Ramachandran) with peptide–protein docking of CD3ζ ITAMs and structural superposition against active ZAP-70 (PDB 4XZ1) to infer residue-level mechanisms for trout Lck1 and ZAP-70.

In Lck1, the residues corresponding to human Y394 and Y505 (trout Y388 and Y499) are predicted to stabilize active and inactive conformations, respectively, supporting a conserved regulatory switch. For ZAP-70, Arg38 and Arg191 are predicted to form the canonical phosphotyrosine-binding pockets required for CD3ζ ITAM engagement. A trout-specific Phe310 at the interdomain interface may favor the autoinhibited state, thereby attenuating kinase activation, suggesting evolutionary fine-tuning. Docking indicated that Lck1 engages the CD3ζ ITAM in a human-like manner, whereas productive ZAP-70 binding required the activation-associated rearrangement seen in 4XZ1, consistent with mammalian mechanisms.

These in silico analyses provide a structural framework for proximal TCR signaling in teleosts. Trout Lck1 harbors the elements needed to initiate ITAM phosphorylation, and ZAP-70 retains key features of ITAM recognition, with Phe310 potentially modulating activation. Experimental validation will be important to test these predictions.

早期t细胞受体（TCR）信号在哺乳动物中被很好地定义，但在虹鳟等硬骨鱼中仍未被充分探索。近端酪氨酸激酶Lck和ZAP-70通过磷酸化和解码CD3 ITAM基序来启动信号传导。在虹鳟鱼中，每个激酶有两个序列的注释；它们的功能相关性尚不清楚。我们将多个序列比对、同源性建模（SWISS-MODEL）和结构质量检查（VERIFY3D, Ramachandran）与CD3ζ ITAMs的肽-蛋白对接和与活性ZAP-70 （PDB 4XZ1）的结构叠加结合起来，推断鳟鱼Lck1和ZAP-70的残留水平机制。在Lck1中，预计与人类Y394和Y505（鳟鱼Y388和Y499）对应的残基分别稳定活性和非活性构象，支持保守的调控开关。对于ZAP-70， Arg38和Arg191被预测形成CD3ζ ITAM结合所需的典型磷酸酪氨酸结合袋。域间界面上的鳟鱼特异性Phe310可能有利于自抑制状态，从而减弱激酶激活，提示进化微调。对接表明Lck1以类似人类的方式结合CD3ζ ITAM，而生产ZAP-70结合需要在4XZ1中看到的激活相关重排，与哺乳动物机制一致。这些计算机分析为硬骨鱼近端TCR信号提供了一个结构框架。鳟鱼Lck1含有启动ITAM磷酸化所需的元件，而ZAP-70保留了ITAM识别的关键特征，Phe310可能调节激活。实验验证对于验证这些预测非常重要。

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引用次数: 0