Fish & shellfish immunology最新文献_第7页

Rab6 mediates lysosome biogenesis to regulate phagocytosis and bacterial clearance in the Chinese mitten crab Eriocheir sinensis Rab6介导中华绒螯蟹溶酶体生物生成调控吞噬和细菌清除。

IF 3.9 2区农林科学 Q1 FISHERIES

Fish & shellfish immunology

Pub Date : 2026-01-16 DOI: 10.1016/j.fsi.2026.111130

Xinyu He , Yuxi Li , Xinyue Wan , Qingding Hao , Yang Li , Guoqing Shen , Kaimin Zhou , Weiwei Li , Qun Wang , Youting Zhu

Phagocytosis by circulating hemocytes is crucial for antibacterial defense in crustaceans, but how intracellular trafficking GTPases coordinate this process remains poorly understood. Here, we identified and characterized a Rab6 ortholog (EsRab6) from the Chinese mitten crab, Eriocheir sinensis, and defined its role in hemocyte phagocytosis and resistance to Vibrio parahaemolyticus. EsRab6 encodes a typical Rab6 GTPase with conserved nucleotide-binding and Rab family motifs and clusters within the invertebrate Rab6 clade. Transcripts were widely expressed, with higher levels in the hepatopancreas and eyestalk, yet EsRab6 was rapidly and transiently upregulated in hemocytes following V. parahaemolyticus challenge. RNA interference-mediated knockdown of EsRab6 resulted in a significant reduction in FITC-labeled bacterial uptake by hemocytes, as shown by microscopy and flow cytometry. Immunofluorescence and LysoTracker staining showed that EsRab6 relocates from a diffuse cytosolic pattern to punctate vesicles that colocalize with lysosomes during infection. Simultaneously, EsRab6 knockdown significantly decreased lysosome-associated signals, indicating a role for EsRab6 in lysosome formation or stability. In vivo, dsRNA-mediated silencing of EsRab6 results in higher hemolymph bacterial loads and significantly lower survival rates following V. parahaemolyticus infection. Overall, these findings identify EsRab6 as a conserved trafficking regulator that links phagosome–lysosome biogenesis to adequate bacterial clearance in crab hemocytes and suggest that Rab6-dependent pathways could be targeted to enhance disease resistance in crustacean aquaculture.

在甲壳类动物中，循环血细胞的吞噬作用对抗菌防御至关重要，但细胞内运输GTPases如何协调这一过程仍然知之甚少。本研究从中华绒螯蟹（Eriocheir sinensis）中鉴定并鉴定了Rab6同源基因EsRab6，并确定了其在血细胞吞噬和对副溶血性弧菌（Vibrio parhaemolyticus）抗性中的作用。EsRab6编码一个典型的Rab6 GTPase，具有保守的核苷酸结合和Rab家族基序，并在无脊椎动物Rab6分支中聚集。转录本广泛表达，在肝胰脏和眼柄中表达水平较高，但在副溶血性弧菌攻击后，EsRab6在血细胞中迅速而短暂地上调。显微镜和流式细胞术显示，RNA干扰介导的EsRab6敲低导致血细胞中fitc标记的细菌摄取显著减少。免疫荧光和LysoTracker染色显示EsRab6在感染期间从弥漫性细胞质模式迁移到与溶酶体共定位的点状囊泡中。同时，EsRab6敲低显著降低溶酶体相关信号，表明EsRab6在溶酶体形成或稳定性中发挥作用。在体内，dsrna介导的EsRab6沉默导致副溶血性弧菌感染后血淋巴细菌负荷升高，生存率显著降低。总的来说，这些发现确定了EsRab6是一种保守的运输调节因子，将吞噬体-溶酶体的生物发生与螃蟹血细胞中足够的细菌清除联系起来，并表明rab6依赖途径可以靶向增强甲壳类水产养殖的抗病能力。

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引用次数: 0

IGF2BP stabilizes RNF34 mRNA to orchestrate apoptosis and host susceptibility to Vibrio splendidus in Apostichopus japonicus IGF2BP稳定RNF34 mRNA调控日本刺参脾弧菌的凋亡和宿主易感性

IF 3.9 2区农林科学 Q1 FISHERIES

Fish & shellfish immunology

Pub Date : 2026-01-14 DOI: 10.1016/j.fsi.2026.111129

Jiqing Liu , Mei Ying , Xuemei Duan , Chenghua Li

Insulin-like growth factor 2 mRNA-binding proteins (IGF2BPs) are conserved N6-methyladenosine (m6A) readers known to orchestrate diverse host immune responses. However, its function in Apostichopus japonicus remains unclear during Vibrio splendidus infection. In this study, we identified a single IGF2BP homolog (AjIGF2BP) in Apostichopus japonicus, which was markedly upregulated following Vibrio splendidus infection. Functional analysis revealed that AjIGF2BP knockdown significantly reduced bacterial load in coelomocytes and mitigated skin ulcer syndrome. Transcriptomic profiling revealed that AjIGF2BP knockdown significantly perturbed apoptotic pathways, among which the E3 ubiquitin ligase ring finger protein 34 (AjRNF34) emerged as a strongly downregulated downstream effector. Mechanistically, AjIGF2BP directly bound to and stabilized AjRNF34 mRNA in an m6A-dependent manner. Rescue experiments confirmed that AjRNF34 was essential for the anti-apoptotic function of AjIGF2BP. Furthermore, AjRNF34 physically interacted with the executioner caspase Ajcaspase-3 via its RING domain and promoted both K48-linked ubiquitination, facilitating its proteasomal degradation and thereby suppressing apoptosis. Collectively, our findings unveil a novel AjIGF2BP/AjRNF34/Ajcaspase-3 regulatory axis through which m6A reader inhibits apoptosis to facilitate bacterial infection, offering new insights into the epigenetic regulatory mechanisms of immunity in echinoderms.

胰岛素样生长因子2 mrna结合蛋白（igf2bp）是保守的n6 -甲基腺苷（m6A）读取器，已知可协调多种宿主免疫反应。然而，在脾弧菌感染期间，其在日本刺参体内的功能尚不清楚。在本研究中，我们在日本带形参（Apostichopus japonicus）中发现了一个单独的IGF2BP同源物（AjIGF2BP），该同源物在脾弧菌感染后显著上调。功能分析显示，AjIGF2BP敲低可显著降低体腔细胞细菌负荷，减轻皮肤溃疡综合征。转录组学分析显示，AjIGF2BP敲低显著干扰凋亡通路，其中E3泛素连接酶环指蛋白34 （AjRNF34）是一个强烈下调的下游效应因子。在机制上，AjIGF2BP以m6a依赖的方式直接结合并稳定AjRNF34 mRNA。救援实验证实AjRNF34对AjIGF2BP的抗凋亡功能至关重要。此外，AjRNF34通过其RING结构域与刽子手caspase Ajcaspase-3物理相互作用，促进k48连接的泛素化，促进其蛋白酶体降解，从而抑制细胞凋亡。总之，我们的研究结果揭示了一个新的AjIGF2BP/AjRNF34/Ajcaspase-3调控轴，m6A读取器通过该调控轴抑制细胞凋亡，促进细菌感染，为棘皮动物免疫的表观遗传调控机制提供了新的见解。

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引用次数: 0

Potent dual-function of marine peptide N6NH2 and its D-enantiomer to combat MDR A. veronii infection in tilapia (GIFT, Oreochromis niloticus) 海洋肽N6NH2及其d -对映体对抗罗非鱼耐多药韦氏杆菌感染的有效双重功能

IF 3.9 2区农林科学 Q1 FISHERIES

Fish & shellfish immunology

Pub Date : 2026-01-14 DOI: 10.1016/j.fsi.2026.111132

Ting Li , Na Yang , Da Teng , Ruoyu Mao , Ya Hao , Huihui Han , Yankang Wu , Xiumin Wang , Jianhua Wang

Tilapia (GIFT, Oreochromis niloticus) has been an economically important freshwater species in China because of its high-quality meat, high yield and high fertility, etc. At present, the high-density breeding industry has suffered great damage from Aeromonas veronii (A. veronii) infections, and the extensive use of antibiotics in aquaculture has resulted in the prevalence of antibiotic-resistant bacteria, resulting in huge economic losses. In this study, we investigated the antibacterial effect of a marine peptide-N6NH₂ and its D-enantiomer (DN6NH₂) on acute A. veronii infections of O. niloticus models, with traditional veterinary antibiotics as controls and found that the DN6NH2 had stronger antibacterial activity in vivo than the parent peptide N6NH₂ and florfenicol. The mortality results showed that DN6NH₂ had more potent efficacy at a dose of 10 mg/kg (81.82 % survival) in O. niloticus peritoneal infection model than 10 mg/kg N6NH₂ (51.52 %) and 10 mg/kg florfenicol (30.30 %). Histopathologic changes observed by HE staining showed significant alleviation in the DN6NH₂ group, inflammation, bleeding and necrosis of the liver, intestine, spleen and gills have been reduced. In addition, in the spleen and head kidney, DN6NH₂ was effective in alleviating the strong immune response to A. veronii infection in O. niloticus by real-time quantitative RT-PCR. In addition, DN6NH₂ significantly reduced nuclear factor-kappaB p65 (NF-κB p65) levels in the spleen after A. veronii infection, indicating its good immunomodulatory capacity. These results suggest the protective action of DN6NH₂ against A. veronii and the potential of this peptide as a promising candidate for aquaculture applications.

罗非鱼（GIFT, Oreochromis niloticus）具有肉质优良、产量高、肥力高等特点，是中国重要的淡水养殖品种。目前，维罗纳气单胞菌（a.w oronii）感染对高密度养殖业造成了很大的损害，而在水产养殖中大量使用抗生素导致耐药菌的流行，造成了巨大的经济损失。本研究以传统兽用抗生素为对照，研究了海洋肽-N6NH2及其d -对映体（DN6NH2）对尼罗ticus急性维罗氏单胞菌感染模型的抑菌作用，发现DN6NH2在体内的抑菌活性强于亲本肽-N6NH2和氟苯尼考。结果表明，DN6NH2在10 mg/kg剂量下对niloticus腹膜感染模型的存活率为81.82%，高于10 mg/kg N6NH2（51.52%）和10 mg/kg氟苯尼考（30.30%）。HE染色观察组织病理学变化，DN6NH2组明显减轻，肝、肠、脾、鳃的炎症、出血、坏死减少。此外，通过实时定量RT-PCR，在脾脏和头肾中，DN6NH2能有效缓解尼罗库蚊对维罗尼阿梭菌感染的强烈免疫反应。此外，DN6NH2显著降低维罗氏单胞杆菌感染后脾脏核因子-κB p65 （NF-κB p65）水平，表明其具有良好的免疫调节能力。这些结果表明DN6NH2对veronii的保护作用以及该肽在水产养殖中的潜在应用前景。

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引用次数: 0

Oyster cellular architecture and innate immune programs resolved by single-cell RNA sequencing 牡蛎细胞结构和先天免疫程序通过单细胞RNA测序解决

IF 3.9 2区农林科学 Q1 FISHERIES

Fish & shellfish immunology

Pub Date : 2026-01-13 DOI: 10.1016/j.fsi.2026.111128

Yi Chen , Wen-Xiong Wang

Barrier mucosal tissues in marine invertebrates coordinate filtration, gas exchange, and frontline innate immunity, yet their cellular organization and division of functions remain poorly resolved. This study generated a tissue-resolved single-cell atlas in a marine invertebrate Crassostrea hongkongensis, defining twenty cell types. The framework revealed a diversified phagocyte lineage composed by five phagocytic cells with three terminal strategies: rapid extracellular containment, deep intracellular clearance, and a resolution program that couples degradation with matrix restoration. We also identified three humoral immune populations with complementary recognition and effector roles that collaborated with phagocytes and epithelial cells. Barrier epithelium supports acted as organizers rather than redundant producers of soluble effectors, and shaped immune activity through defined communication and adhesion programs that coordinated proliferation, survival, and migration. A neural-related compartment comprised of neural progenitor-like, neuron-like, and neuroendocrine cells, and exhibited predominantly sender roles toward epithelial and immune targets consistent with transmitter and peptide-mediated control of ciliary flow, secretory activity, as well as the priming of phagocytic and humoral effectors. Ligand receptor inference highlighted adhesion modules centering on neural cell adhesion molecule (NCAM) and the receptor type protein tyrosine phosphatase (PTPRM) as the key pathways of communication between neural, epithelial, and immune populations. Our finding present a compact taxonomy to map immune programs with precise cellular contexts, and further our understanding of resilience and health in variable coastal environments.

海洋无脊椎动物的屏障粘膜组织协调过滤、气体交换和一线先天免疫，但其细胞组织和功能划分仍不清楚。本研究生成了一种海洋无脊椎动物香港长牡蛎的组织分辨单细胞图谱，定义了20种细胞类型。该框架揭示了由五种吞噬细胞组成的多样化的吞噬细胞谱系，它们具有三种终端策略：快速的细胞外遏制，细胞内深度清除和降解与基质恢复相结合的解决方案。我们还确定了三种具有互补识别和效应作用的体液免疫群体，它们与吞噬细胞和上皮细胞协同作用。屏障上皮支持作为可溶性效应物的组织者而不是多余的生产者，并通过明确的沟通和粘附程序来协调增殖、存活和迁移，从而形成免疫活性。一种由神经祖细胞样细胞、神经元样细胞和神经内分泌细胞组成的神经相关细胞室，主要表现出对上皮和免疫目标的发送者作用，与睫状体流动、分泌活性以及吞噬和体液效应的递质和肽介导的控制一致。配体受体推断强调以神经细胞粘附分子（NCAM）和受体型蛋白酪氨酸磷酸酶（PTPRM）为中心的粘附模块是神经、上皮和免疫群体之间沟通的关键途径。我们的发现提供了一个紧凑的分类学，以精确的细胞背景绘制免疫程序，并进一步了解我们在可变沿海环境中的恢复力和健康。

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引用次数: 0

Establishment of a caudal fin cell line from largemouth bass (Micropterus salmoides) and its transcriptomic response to largemouth bass virus infection 大口黑鲈尾鳍细胞系的建立及其对大口黑鲈病毒感染的转录组反应

IF 3.9 2区农林科学 Q1 FISHERIES

Fish & shellfish immunology

Pub Date : 2026-01-12 DOI: 10.1016/j.fsi.2026.111122

Jingting Qi , Wenting Wang , Yimeng Liu , Qiao Huang , Mingli Liu , Chaoguang Wei , Yuang Jiang , Zhichao Wu , Zhitao Qi , Jiulin Chan , Peng Hu

The largemouth bass (Micropterus salmoides), commonly known as the California bass, is a species of significant economic importance in global freshwater aquaculture. Largemouth bass virus (LMBV) has emerged as a major pathogen that poses a substantial threat to the sustainability of largemouth bass aquaculture. In this study, we established a novel Micropterus salmoides caudal fin (MSCF) cell line that supports LMBV propagation and enables investigation of virus-induced fundamental patterns of molecular responses. Characterization of the MSCF line revealed an initial mixture of epithelial-like and fibroblast-like cells that became uniformly epithelial by passage 20. Transcriptomic analysis revealed that LMBV infection significantly altered the transcription levels of 262 genes (201 up-regulated genes and 61 down-regulated), which were functionally enriched in key antiviral pathways such as apoptosis, FoxO signaling, and RIG-I-like receptor signaling. The observed expression patterns suggest that LMBV employs a potential strategy to manipulate host cell survival and disrupt innate immune responses. Collectively, we have developed an LMBV-susceptible cell line (MSCF), which provides an ideal platform for studying LMBV virology and elucidating the host defense mechanisms against LMBV.

大口黑鲈（Micropterus salmoides），俗称加州黑鲈，是全球淡水水产养殖中具有重要经济意义的一种。大口黑鲈病毒（LMBV）已成为威胁大口黑鲈养殖可持续性的主要病原体。在这项研究中，我们建立了一种支持LMBV繁殖和研究病毒诱导的分子反应基本模式的新型鲑鱼小翼（Micropterus salmoides尾鳍）细胞系。MSCF细胞系的特征显示，在传代第20代时，上皮样细胞和成纤维细胞样细胞的初始混合物成为均匀的上皮细胞。转录组学分析显示，LMBV感染显著改变了262个基因的转录水平（201个上调基因，61个下调基因），这些基因在凋亡、FoxO信号和rig - i样受体信号等关键抗病毒途径中功能丰富。观察到的表达模式表明，LMBV采用了一种潜在的策略来操纵宿主细胞存活并破坏先天免疫反应。我们共同开发了LMBV易感细胞系（MSCF），为研究LMBV病毒学和阐明宿主对LMBV的防御机制提供了理想的平台。

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引用次数: 0

Combined histopathological, immunoenzymatic and transcriptomic analyses reveal the immune response mechanisms of silver pomfret infected by Vibrio parahaemolyticus 结合组织病理学、免疫酶学和转录组学分析，揭示了副溶血性弧菌感染鲳鱼的免疫反应机制

IF 3.9 2区农林科学 Q1 FISHERIES

Fish & shellfish immunology

Pub Date : 2026-01-12 DOI: 10.1016/j.fsi.2026.111124

Dianyang Zhou , Ping Han , Yadong Xue , Zhennan Sun , Longdi Wang , Xiumei Liu , Jianming Chen , Yajun Wang , Suming Zhou , Xue Sun , Xinxin Du , Xubo Wang

The silver pomfret (Pampus argenteus), a vital aquaculture species, is highly susceptible to Vibrio parahaemolyticus infections, yet the molecular mechanisms underlying its immune response remain poorly understood. This study investigated gill responses to V. parahaemolyticus infection via intraperitoneal injection, analyzing histopathological, transcriptomic, and enzymatic changes at 3, 6, 12, 24 and 48 h post-infection (hpi). Histological analysis revealed progressive gill damage, with significant architectural disruption from 6 hpi and severe lesions by 48 hpi. Transcriptome profiling of 18 cDNA libraries generated 40–67 million clean reads, yielding 4492–5231 differentially expressed genes (DEGs) across time points, with 1094–2174 unique DEGs per time point. Inflammatory cytokines (tnf-α, il-1β, il-8, il-12) showed robust upregulation, peaking at 6–24 hpi, indicative of an active immune response. Gene Ontology (GO) and KEGG analyses identified enriched pathways, including “oxidative phosphorylation” (3–48 hpi) and “tight junction” (24 hpi), with “oxidoreductase activity” and “aerobic respiration” as the dominant functional categories. Oxidative stress markers (MDA, SOD, CAT, POD) exhibited time-dependent increases, reflecting enhanced antioxidant defenses. Weighted Gene Co-expression Network Analysis (WGCNA) revealed the turquoise module as being strongly correlated with infection, identifying hub genes (gapdh, gapdhs, pgam2, g6pd, taldo1, pgam1a) critical to immune regulation. This study is the first to systematically investigate the temporal crosstalk between inflammatory responses, oxidative stress, and metabolic reprogramming in silver pomfret gills during V. parahaemolyticus infection, providing novel molecular targets for the control of vibriosis.

银鲳鱼（Pampus argteus）是一种重要的水产养殖物种，对副溶血性弧菌感染非常敏感，但其免疫反应的分子机制尚不清楚。本研究通过腹腔注射研究了鳃对副溶血性弧菌感染的反应，分析了感染后3、6、12、24和48小时（hpi）的组织病理学、转录组学和酶学变化。组织学分析显示进行性鳃损伤，6 hpi时出现明显的结构破坏，48 hpi时出现严重病变。18个cDNA文库的转录组分析产生了4000 - 6700万个干净reads，在不同的时间点上产生4492-5231个差异表达基因（DEGs），每个时间点有1094-2174个独特的DEGs。炎症因子（tnf-α, il-1β, il-8, il-12）表现出强劲的上调，在6-24 hpi时达到峰值，表明积极的免疫反应。基因本体（GO）和KEGG分析确定了富集的途径，包括“氧化磷酸化”（3-48 hpi）和“紧密连接”（24 hpi），“氧化还原酶活性”和“有氧呼吸”是主要的功能类别。氧化应激标志物（MDA、SOD、CAT、POD）呈时间依赖性增加，反映抗氧化防御能力增强。加权基因共表达网络分析（WGCNA）显示，绿松石模块与感染密切相关，确定了对免疫调节至关重要的枢纽基因（gapdh、gapdhs、pgam2、g6pd、taldo1、pgam1a）。本研究首次系统地研究了副溶血性弧菌感染期间鲳鱼鳃炎症反应、氧化应激和代谢重编程之间的时间串聊，为控制弧菌病提供了新的分子靶点。

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引用次数: 0

The E3 ligase RNF5 facilitates viral replication by suppressing innate immune responses in grouper E3连接酶RNF5通过抑制石斑鱼的先天免疫反应促进病毒复制

IF 3.9 2区农林科学 Q1 FISHERIES

Fish & shellfish immunology

Pub Date : 2026-01-12 DOI: 10.1016/j.fsi.2026.111123

Yanqi Zhao , Xiaohui Zheng , Siting Wu , Minyao Bi , Jiaming Hu , Shaojie Liu , Jingguang Wei , Qiwei Qin

Singapore grouper iridovirus (SGIV) and red-spotted grouper nervous necrosis virus (RGNNV) are major pathogens that cause high mortality and substantial economic losses in grouper aquaculture. The role of ring finger protein 5 (RNF5), an E3 ubiquitin ligase, in the antiviral innate immunity of teleosts remains poorly understood. In this study, a homolog of the RNF5 gene was identified and functionally characterized from Epinephelus coioides, which was designated EcRNF5. The open reading frame (ORF) of EcRNF5 is 645 nucleotides in length, encoding a 214-amino acid protein that contains a characteristic RING domain. Phylogenetic analysis revealed that EcRNF5 shares the closest evolutionary relationship with its ortholog in Epinephelus fuscoguttatus. EcRNF5 was ubiquitously expressed in all examined tissues. Furthermore, its transcription level was significantly upregulated in grouper spleen (GS) cells following challenge with SGIV or RGNNV. Overexpression of EcRNF5 enhanced SGIV and RGNNV replication in vitro. Concurrently, it suppressed the expression of interferon-related genes and pro-inflammatory cytokines. The promoter activities of interferon 3 (IFN3), interferon-stimulated response element (ISRE), and nuclear factor kappa B (NF-κB) were also inhibited. In addition, EcRNF5 attenuated IFN3 promoter activation induced by the key signaling molecules: EcSTING, EcTBK1, EcIRF3, and EcIRF7. Co-immunoprecipitation (Co-IP) assays confirmed physical interactions between EcRNF5 and these four innate immune signaling molecules. Collectively, these findings provide novel insights into the functional role of RNF5 in fish-virus interactions and shed light on the molecular mechanisms underlying SGIV and RGNNV pathogenicity in grouper.

新加坡石斑鱼虹膜病毒（SGIV）和红斑石斑鱼神经坏死病毒（RGNNV）是造成石斑鱼养殖高死亡率和重大经济损失的主要病原体。环指蛋白5 (RNF5)，一种E3泛素连接酶，在硬骨鱼抗病毒先天免疫中的作用仍然知之甚少。本研究从石斑鱼中鉴定了一个RNF5同源基因，并对其进行了功能鉴定，命名为EcRNF5。EcRNF5的开放阅读框（ORF）长度为645个核苷酸，编码214个氨基酸的蛋白，包含一个特征环结构域。系统发育分析表明，EcRNF5在褐纹石斑鱼中与其同源基因具有最密切的进化关系。EcRNF5在所有检测组织中普遍表达。此外，SGIV或RGNNV攻击后，其转录水平在石斑鱼脾脏（GS）细胞中显著上调。过表达EcRNF5可增强SGIV和RGNNV的体外复制。同时抑制干扰素相关基因和促炎细胞因子的表达。干扰素3 （IFN3）、干扰素刺激反应元件（ISRE）和核因子κB （NF-κB）的启动子活性也受到抑制。此外，EcRNF5减弱了关键信号分子EcSTING、EcTBK1、EcIRF3和EcIRF7诱导的IFN3启动子激活。共免疫沉淀（Co-IP）实验证实了EcRNF5与这四种先天免疫信号分子之间的物理相互作用。总的来说，这些发现为RNF5在鱼与病毒相互作用中的功能作用提供了新的见解，并揭示了石斑鱼SGIV和RGNNV致病性的分子机制。

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引用次数: 0

Surface-displayed MCP-3 vaccine in Bacillus subtilis elicits protective immune responses against LMBV infection in largemouth bass 表面显示的枯草芽孢杆菌MCP-3疫苗引起了大口黑鲈对LMBV感染的保护性免疫反应。

IF 3.9 2区农林科学 Q1 FISHERIES

Fish & shellfish immunology

Pub Date : 2026-01-12 DOI: 10.1016/j.fsi.2026.111126

Ruiqi Lin , Zhihao Jiang , Qianqian Zhang , Shun Li , Xueying Qin , Xinyou Wang , Gaofeng Cheng , Zhen Xu , Weiguang Kong

Largemouth bass virus (LMBV) is a major pathogen threatening the aquaculture of largemouth bass, causing significant economic losses. Oral vaccination offers a practical and efficient strategy for disease prevention in farmed fish. In this study, a fusion CotC-MCP-3 gene was introduced into Bacillus subtilis 168 (Bs168) to enable surface display of the MCP-3 protein on bacterial spores through the anchoring protein CotC, thereby generating a candidate oral vaccine. At 28 days post-vaccination (dpv), qPCR analysis revealed pronounced upregulation of associated innate and adaptive immunity genes in the gut and head kidney of largemouth bass immunized with the MCP-3 vaccine. Vaccination markedly increased the abundance of IgM⁺ B cells in both the gut and head kidney, as well as total IgM levels in serum and gut mucus. Concomitantly, LMBV-specific IgM titers and neutralizing activities in both serum and gut mucus were significantly enhanced. Following the experimental LMBV challenge, fish in the MCP-3 vaccine group achieved a relative percent survival (RPS) of 68.7 %, accompanied by substantially lower viral loads in the head kidney, gut, and spleen, as well as notably attenuated histopathological lesions compared with the other groups. The Bs168-based MCP-3 oral vaccine provides strong protection against LMBV infection in largemouth bass, as demonstrated by these results, highlighting its potential as a promising strategy for managing viral diseases in aquaculture.

大口黑鲈病毒（Largemouth bass virus， LMBV）是威胁大口黑鲈养殖的主要病原体，造成了重大的经济损失。口服疫苗接种为养殖鱼提供了一种实用而有效的疾病预防策略。本研究将融合的CotC-MCP-3基因导入枯草芽孢杆菌168 （Bs168）中，使MCP-3蛋白通过锚定蛋白CotC在细菌孢子表面展示，从而产生候选口服疫苗。接种MCP-3疫苗后28天，qPCR分析显示，接种MCP-3疫苗的大口黑鲈肠道和头肾中相关先天免疫和适应性免疫基因明显上调。疫苗接种显著增加了肠道和头肾中IgM+ B细胞的丰度，以及血清和肠道粘液中IgM的总水平。同时，血清和肠道粘液中lmbv特异性IgM滴度和中和活性显著增强。在实验性LMBV攻击后，MCP-3疫苗组的鱼获得了68.7%的相对存活率（RPS），与其他组相比，头部肾脏、肠道和脾脏的病毒载量大大降低，组织病理学病变也明显减轻。这些结果表明，基于bs168的MCP-3口服疫苗对大口黑鲈的LMBV感染提供了强有力的保护，突出了其作为一种有前途的水产养殖病毒性疾病管理策略的潜力。

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引用次数: 0

Yorkie antagonizes innate antiviral immunity through regulating the IRF and Dorsal mediated antiviral pathways in Litopenaeus vannamei Yorkie通过调节IRF和背侧介导的抗病毒途径拮抗凡纳滨对虾的先天抗病毒免疫

IF 3.9 2区农林科学 Q1 FISHERIES

Fish & shellfish immunology

Pub Date : 2026-01-12 DOI: 10.1016/j.fsi.2026.111121

Guoqing Dai , Wei Xiao , Huiling Xing , Lili Shi

Yorkie

(Yki), a key effector of the Hippo signaling pathway, plays important roles in the regulation of cell proliferation and immune responses. In this study, two splice variants of the Yorkie gene (designated as LvYki-long and LvYki-short, respectively) were identified from Litopenaeus vannamei. The open reading frame (ORF) of LvYki-long is 1650 bp that encodes a putative protein of 549 amino acids. Compared with the amino acid sequence of LvYki-long, the amino acid sequence of LvYki-short is reduced by 78 amino acids (positions 416–493). Both LvYki-long and LvYki-short were universally expressed in all tested tissues. Following WSSV and DIV1 infection, their expression was significantly up-regulated in hemocytes and hepatopancreas but down-regulated in gills. The dsRNA-mediated knockdown of LvYki substantially enhanced the shrimp survival rate and reduced viral loads after WSSV and DIV1 infection. Besides, silencing of LvYki led to the down-regulation of Cactus, the up-regulation of Dorsal, as well as several AMPs (ALF1, ALF2, ALF4, and SWD3). Furthermore, silencing of LvYki also resulted in increased expression levels of Vago-JAK/STAT pathway components (Vago4, Vago5, and STAT), and the Co-IP results showed that LvYki could interact with LvIRF. The dual luciferase assay verified that LvYki significantly inhibited the activation of the Vago4 promoter by LvIRF. These results indicated that LvYki might antagonize the innate antiviral immunity through regulating the IRF and Dorsal-mediated antiviral pathways.

Yorkie（Yki）是Hippo信号通路的关键效应蛋白，在细胞增殖和免疫应答的调控中发挥重要作用。本研究从凡纳滨对虾（Litopenaeus vannamei）中鉴定出两个Yorkie基因剪接变体（分别命名为LvYki-long和LvYki-short）。LvYki-long的开放阅读框（ORF）长度为1650 bp，编码549个氨基酸的推定蛋白。与LvYki-long的氨基酸序列相比，LvYki-short的氨基酸序列减少了78个氨基酸（第416 ~ 493位）。绿脉基长型和绿脉基短型均在所有组织中普遍表达。WSSV和DIV1感染后，其在血细胞和肝胰腺中的表达显著上调，而在鳃中的表达明显下调。dsrna介导的LvYki基因敲低显著提高了WSSV和DIV1感染后虾的存活率，降低了病毒载量。此外，LvYki的沉默导致Cactus下调，Dorsal上调，以及几种amp （ALF1、ALF2、ALF4、SWD3）上调。此外，LvYki的沉默还导致Vago-JAK/STAT通路组分（Vago4、Vago5和STAT）的表达水平升高，Co-IP结果表明LvYki可以与LvIRF相互作用。双荧光素酶实验证实LvYki显著抑制了LvIRF对Vago4启动子的激活。这些结果表明绿毒素可能通过调节IRF和dorsal介导的抗病毒途径拮抗先天抗病毒免疫。

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引用次数: 0

Recombinant phage-display monoclonal antibody against starry flounder (Platichthys stellatus) IgM enables quantitative ELISA for vaccine-induced humoral responses 重组星形比目鱼（Platichthys stellatus） IgM噬菌体展示单克隆抗体可用于疫苗诱导的体液反应的定量ELISA检测

IF 3.9 2区农林科学 Q1 FISHERIES

Fish & shellfish immunology

Pub Date : 2026-01-12 DOI: 10.1016/j.fsi.2026.111127

Min-Young Sohn , Jun-ichi Hikima , Gyoungsik Kang , Kyung-Ho Kim , Ha-Jeong Son , Chan-Il Park

Species-matched serological reagents are scarce for many aquaculture species, limiting quantitative assessment of vaccine responses. Here, we developed and validated monoclonal antibodies (MAbs) specific to starry flounder (Platichthys stellatus) immunoglobulin M (IgM) using a recombinant phage-display approach. IgM purified from immune serum was used as the antigen for bio-panning, yielding multiple IgM-reactive scFv candidates, all of which recognized the IgM heavy chain by Western blot analysis. A lead clone (1T1A11) was reformatted as an scFv-Fc minibody, expressed in mammalian cells, purified by Protein A affinity chromatography, and verified as a single band of the expected molecular size. The 1T1A11 minibody specifically immunoprecipitated IgM from both purified preparations and serum, confirming its functional specificity. Using this monoclonal antibody, we established an optimized indirect ELISA to quantitatively monitor humoral immune responses following Streptococcus parauberis formalin-killed cell vaccination. The assay revealed clear dose- and time-dependent increases in serum IgM levels, which were further quantified using a purified starry flounder IgM standard curve. Collectively, these results establish 1T1A11 as a specific and scalable reagent that enables standardized, quantitative serological analysis for vaccine evaluation and diagnostic applications in flatfish aquaculture.

许多水产养殖品种缺乏与品种匹配的血清学试剂，限制了疫苗反应的定量评估。在这里，我们利用重组噬菌体展示方法开发并验证了星牙鲆（Platichthys stellatus）免疫球蛋白M （IgM）的单克隆抗体。从免疫血清中纯化IgM作为抗原进行生物筛选，得到多个IgM反应性scFv候选体，经Western blot分析均识别IgM重链。一个先导克隆（1T1A11）被重组为scFv-Fc小体，在哺乳动物细胞中表达，通过蛋白A亲和层析纯化，并被验证为预期分子大小的单带。1T1A11小体从纯化制剂和血清中特异性免疫沉淀IgM，证实了其功能特异性。利用该单克隆抗体，建立了一种优化的间接ELISA定量监测副金黄色葡萄球菌福尔马林杀伤细胞接种后的体液免疫应答。该实验显示血清IgM水平明显呈剂量和时间依赖性增加，使用纯化的星形比目鱼IgM标准曲线进一步定量。总的来说，这些结果确立了1T1A11是一种特异性和可扩展的试剂，可用于比目鱼养殖中疫苗评估和诊断应用的标准化、定量血清学分析。

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引用次数: 0