Fish & shellfish immunology最新文献_第8页

Evaluation of recombinant SIP, enolase, and GAPDH as subunit vaccine candidates against Streptococcus iniae in four-finger threadfin (Eleutheronema tetradactylum): Immunogenicity, protective efficacy, and multivalent potential 重组SIP、烯醇酶和GAPDH作为四指线虫猪链球菌亚单位候选疫苗的评价：免疫原性、保护效果和多价潜力

IF 3.9 2区农林科学 Q1 FISHERIES

Fish & shellfish immunology

Pub Date : 2026-01-21 DOI: 10.1016/j.fsi.2026.111146

Andre Giovanni , Yin-Ze Shi , Pei-Chi Wang , Ming-An Tsai , Shih-Chu Chen

Streptococcus iniae is a highly virulent pathogen causing severe streptococcosis outbreaks and high mortality in cultured fish, including four-finger threadfin fish in Taiwan. This study evaluated the immunogenicity and protective efficacy of recombinant enolase, surface immunogenic protein (SIP), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH), administered individually and in combination, as subunit vaccines against S. iniae infection. Recombinant proteins were expressed in Escherichia coli, purified, and emulsified in an oil-based adjuvant before intraperitoneal vaccination of threadfin fish. Immune responses were assessed through serum IgM levels, lysozyme activity, immune gene expression, serum bactericidal activity, and relative percent survival (RPS) after challenge with virulent S. iniae. The results demonstrated successful expression of target proteins and significant elevation of antigen-specific IgM antibody level, particularly in the combination vaccine group. Enhanced lysozyme activity and upregulation of complement component C2, T cell marker CD8, and cytokines (TNF-α, IL-12, IL-10) indicated robust innate and adaptive immune activation. Serum bactericidal assays revealed the highest bacterial killing in fish vaccinated with the combination formulation. Post-challenge survival was greatest in fish receiving formalin-killed cell vaccine (RPS 86.2 %), followed by moderate protection conferred by recombinant SIP (rSIP) (72.4 %) and the combination vaccine (52.7 %). Recombinant enolase and GAPDH provided lower protection levels. These findings highlight the potential of rSIP and multivalent subunit vaccines as promising strategies for controlling S. iniae infections in aquaculture.

牛链球菌是一种高毒力病原体，可在养殖鱼类中引起严重的链球菌病爆发和高死亡率，包括台湾的四指带鱼。本研究评估了重组烯醇化酶、表面免疫原性蛋白（SIP）和甘油醛-3-磷酸脱氢酶（GAPDH）单独或联合作为猪链球菌感染亚单位疫苗的免疫原性和保护效果。重组蛋白在大肠杆菌中表达，纯化，并在油基佐剂中乳化，然后腹腔注射长针鱼。免疫应答通过血清IgM水平、溶菌酶活性、免疫基因表达、血清杀菌活性和强毒链球菌攻击后的相对存活率（RPS）来评估。结果显示靶蛋白的成功表达和抗原特异性IgM抗体水平的显著升高，特别是在联合疫苗组。溶菌酶活性的增强和补体成分C2、T细胞标志物CD8和细胞因子（TNF-α、IL-12、IL-10）的上调表明了强大的先天和适应性免疫激活。血清杀菌试验显示，接种联合制剂的鱼的细菌杀灭率最高。接受福尔马林杀伤细胞疫苗的鱼的攻毒后存活率最高（RPS为86.2%），其次是重组SIP (rSIP)（72.4%）和联合疫苗（52.7%）。重组烯醇化酶和GAPDH提供较低的保护水平。这些发现突出了rSIP和多价亚单位疫苗作为控制水产养殖中猪链球菌感染的有希望的策略的潜力。

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引用次数: 0

Hematology-centered physiological remodeling of black seabream (Acanthopagrus schlegelii) under gradual water temperature fluctuations 以血液学为中心的黑鲷在水温逐渐波动下的生理重塑

IF 3.9 2区农林科学 Q1 FISHERIES

Fish & shellfish immunology

Pub Date : 2026-01-21 DOI: 10.1016/j.fsi.2026.111150

Seok-Ju Lee , Su-Mi Shin , Jae-Hyun Sim , Da-Jeong Yoo , Eunju Kim , Myung-Hwa Jung

Water temperature profoundly shapes the physiology and immunity of poikilothermic fish, yet gradual thermal transitions remain poorly characterized. This study examined hematological, biochemical, and gene-expression responses derived entirely from whole blood of black seabream (Acanthopagrus schlegelii) exposed to progressive warming (30 °C) and cooling (8 °C). Short-term thermal response proceeded through reversible phases of activation, remodeling, and stabilization as reflected in hematological, biochemical, and molecular indices. Moderate cooling (20–18 °C) induced the most dynamic reorganization, including transient increases in monocytes and granulocytes, elevated alanine aminotransferase activity, and lipid depletion indicating early innate immune deployment and metabolic redistribution preceding deep cold suppression. Several hematological parameters exhibited nonlinear, compensatory trends: mean corpuscular hemoglobin concentration (MCHC) increased under cold exposure, whereas red cell distribution width (RDW) expanded at both thermal extremes, revealing alternative strategies for maintaining oxygen transport and erythrocyte stability. Gene expression showed selective and pathway-specific regulation, with interleukin1beta (IL1β) up-regulated at both water temperature extremes, heat shock protein 90 (HSP90) elevated under heat stress, and glutathione peroxidase 1 (GPX1) down-regulated under cold exposure, reflecting flexible and individualized physiological adjustments. Taken together, these findings establish a blood-centered model of short-term thermal response, in which whole-blood physiology functions as a regulatory hub integrating oxygen transport, immune modulation, and metabolic balance. These findings highlight blood parameters as sensitive, non-invasive biomarkers of short-term thermal response, particularly revealing a previously underappreciated moderate-cooling phase that drives immune and metabolic reorganization.

水温深刻地塑造了变热鱼类的生理和免疫，但逐渐的热转变仍然很不清楚。本研究检测了暴露于逐渐升温（30°C）和冷却（8°C）的黑海鲷（棘海鲷）全血的血液学、生化和基因表达反应。短期热反应经历了激活、重塑和稳定的可逆阶段，这反映在血液学、生化和分子指标上。适度冷却（20-18°C）诱导了最动态的重组，包括单核细胞和粒细胞的短暂增加，丙氨酸转氨酶活性升高，脂质消耗表明在深度冷抑制之前早期先天免疫部署和代谢重新分配。几个血液学参数表现出非线性的补偿趋势：平均红细胞血红蛋白浓度（MCHC）在冷暴露下增加，而红细胞分布宽度（RDW）在两个极端温度下都扩大，揭示了维持氧运输和红细胞稳定性的替代策略。基因表达表现出选择性和途径特异性调控，白介素1β (il - 1β)在两种极端水温条件下均上调，热休克蛋白90 （HSP90）在热应激条件下升高，谷胱甘肽过氧化物酶1 （GPX1）在冷暴露条件下下调，反映出灵活和个性化的生理调节。综上所述，这些发现建立了一个以血液为中心的短期热反应模型，其中全血生理学作为一个整合氧运输、免疫调节和代谢平衡的调节枢纽。这些发现强调了血液参数是短期热反应的敏感、非侵入性生物标志物，特别是揭示了以前未被重视的驱动免疫和代谢重组的中度冷却阶段。

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引用次数: 0

Toxic effects of microplastic (polyethylene) exposure: Stress, immune responses and neurotoxicity in crucian carp, Carassius carassius 微塑料（聚乙烯）暴露的毒性效应：应激、免疫反应和鲫鱼的神经毒性。

IF 3.9 2区农林科学 Q1 FISHERIES

Fish & shellfish immunology

Pub Date : 2026-01-20 DOI: 10.1016/j.fsi.2026.111139

Young-Bin Yu , A-Hyun Jo , Cheol Young Choi , Ju-Chan Kang , Jun-Hwan Kim

Crucian carp (Carassius carassius) were exposed to polyethylene microplastics (PE-MPs) at concentrations of 0, 4, 8, 16, 32, and 64 mg/L for two weeks. The experimental fish had a mean weight of 23.2 ± 3.1 g and a mean length of 12.1 ± 0.9 cm. Exposure to ≥32 mg/L PE-MPs caused significant increases in cortisol and HSP70 levels, indicating activation of endocrine and cellular stress responses. In contrast, immune parameters such as lysozyme activity and IgM concentrations were significantly reduced, demonstrating suppression of both innate and adaptive immune functions. Moreover, acetylcholinesterase (AChE) activity was significantly inhibited, reflecting impaired cholinergic neurotransmission and neurotoxicity. Integrated biomarker response (IBR) analysis revealed concentration-dependent alterations, while heatmap and principal component analyses (PCA) confirmed that PE-MPs predominantly induced stress-related physiological responses. Collectively, these findings demonstrate that exposure to ≥32 mg/L PE-MPs poses significant physiological and neuro-immunotoxic risks to C. carassius.

以0、4、8、16、32和64 mg/L浓度的聚乙烯微塑料（PE-MPs）对鲫鱼（Carassius Carassius）进行为期2周的暴露试验。实验鱼的平均体重为23.2±3.1 g，平均体长为12.1±0.9 cm。暴露于≥32 mg/L PE-MPs导致皮质醇和HSP70水平显著升高，表明内分泌和细胞应激反应被激活。相比之下，溶菌酶活性和IgM浓度等免疫参数显著降低，表明先天和适应性免疫功能均受到抑制。此外，乙酰胆碱酯酶（AChE）活性明显受到抑制，反映出胆碱能神经传递和神经毒性受损。综合生物标志物反应（IBR）分析显示了浓度依赖性的变化，而热图和主成分分析（PCA）证实PE-MPs主要诱导应激相关的生理反应。综上所述，这些发现表明暴露于≥32 mg/L PE-MPs对鲫鱼具有显著的生理和神经免疫毒性风险。

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引用次数: 0

Midnolin has an ancient origin and negatively regulates type I IFN response in teleost fish Midnolin具有古老的起源并负向调节硬骨鱼I型IFN反应。

IF 3.9 2区农林科学 Q1 FISHERIES

Fish & shellfish immunology

Pub Date : 2026-01-20 DOI: 10.1016/j.fsi.2026.111134

Yuan Feng , Wenxing Li , Jiamei Liu , Huazhi Chen , Pan Qin , Jingqun Ao , Xinhua Chen

Midnolin has recently been shown to mediate ubiquitin-independent proteasomal degradation of nuclear substrates, yet its evolutionary origin and function in non-mammalian immunity remain largely unexplored. Here, we identify a Midnolin homolog in large yellow croaker (Larimichthys crocea). A key finding is that the origin of Midnolin can be traced back to placozoans, the simplest known animals whose emergence predates that of sponges. In L. crocea, Lcmidnolin expression was significantly upregulated in immune-related tissues upon stimulation with poly(I:C), a viral mimic, suggesting its responsiveness to viral-like signals. Confocal microscopy revealed nuclear localization of LcMidnolin. Functionally, overexpression of LcMidnolin suppressed poly(I:C)-induced activation of type I interferon (IFN) promoters and diminished antiviral gene expression. Consistent with this inhibitory effect, LcMidnolin overexpression impaired the cellular antiviral response, as reflected by increased viral gene expression and exacerbated cytopathic effects in cells infected with spring viremia of carp virus (SVCV). Our findings provide the first evolutionary and functional insights into Midnolin in a non-mammalian vertebrate, revealing its previously unrecognized role as a regulator of the type I IFN signaling pathway.

Midnolin最近被证明介导核底物的泛素非依赖性蛋白酶体降解，但其进化起源和在非哺乳动物免疫中的功能仍未被广泛探索。本文中，我们在大黄鱼（Larimichthys crocea）中鉴定了Midnolin同源物。一个关键的发现是，Midnolin的起源可以追溯到盘虫，这是已知最简单的动物，其出现早于海绵。在L. crocea中，Lcmidnolin在受到病毒模拟物poly（I:C）刺激后，在免疫相关组织中的表达显著上调，表明其对病毒样信号的反应性。共聚焦显微镜显示LcMidnolin的核定位。在功能上，LcMidnolin的过表达抑制了poly（I:C）诱导的I型干扰素（IFN）启动子的激活和抗病毒基因表达的减少。与这种抑制作用一致的是，LcMidnolin过表达损害了细胞抗病毒反应，这反映在感染鲤鱼病毒春季病毒血症（SVCV）的细胞中病毒基因表达增加和细胞病变效应加剧。我们的研究结果首次揭示了Midnolin在非哺乳动物脊椎动物中的进化和功能，揭示了它作为I型IFN信号通路调节因子的作用。

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引用次数: 0

Development and identification of KASP-SNP markers correlated with Aeromonas hydrophila resistance traits in blunt snout bream (Megalobrama amblycephala) 钝口鲷（Megalobrama amblycephala）嗜水气单胞菌抗性相关KASP-SNP标记的开发与鉴定。

IF 3.9 2区农林科学 Q1 FISHERIES

Fish & shellfish immunology

Pub Date : 2026-01-20 DOI: 10.1016/j.fsi.2026.111144

Han Wang , Chun Liu , Anicet Philippe Mane Sany , Ziquan Yang , Jie Chen , Guodong Zheng , Shuming Zou

The blunt snout bream (Megalobrama amblycephala) is an economically important freshwater fish species. However, it is highly susceptible to Aeromonas hydrophila infection, especially in intensive pond aquaculture in China. Molecular marker-assisted selection provides an efficient approach for breeding disease-resistant varieties; however, the key genes or molecular markers linked to A. hydrophila resistance remain scarce in this species. A 436 differential SNP sites with disease-resistant were screened on basis of whole-genome resequencing. Then, a high-throughput genomic KASP genotyping technique was utilized to discover favorable genes and SNP sites associated with A. hydrophila resistance. A total of 46 KASP markers were successfully developed with an accuracy of 92 %. These markers were used to genotyping 120 blunt snout bream individuals. Through trait correlation analysis and general linear models (GLM), five SNPs significantly (P < 0.05) associated with resistance to A. hydrophila were identified and mapped to five candidate genes (btnl2, cfhr2, slc47a1, neu3, nlrp1). Survival rate of individuals carrying the dominant genotype demonstrated an average survival rate of 81.39 %, which represents a 69.35 % increase in comparison with that of 48 % in total population. This effect was validated in an external population of 100 fish. These findings identify key genetic markers associated with A. hydrophila resistance and provide a direction for elucidating the underlying molecular immune mechanisms, thus establishing a genetic foundation for future breeding strategies.

钝口鲷（Megalobrama amblycephala）是一种经济上重要的淡水鱼种。然而，在中国的集约化池塘养殖中，它极易感染嗜水气单胞菌。分子标记辅助选择为抗病品种的选育提供了有效途径；然而，与嗜水单胞杆菌抗性相关的关键基因或分子标记在该物种中仍然缺乏。在全基因组重测序的基础上，筛选了436个具有抗病的差异SNP位点。然后，利用高通量基因组KASP基因分型技术，发现与嗜水单胞杆菌耐药性相关的有利基因和SNP位点。共建立了46个KASP标记，准确率为92%。利用这些标记对120条钝口鲷进行了基因分型。通过性状相关分析和一般线性模型（GLM），鉴定出5个与嗜水单胞菌抗性显著（P < 0.05）相关的snp，并定位到5个候选基因（btnl2、cfhr2、slc47a1、neu3、nlrp1）。携带显性基因型个体的平均存活率为81.39%，比种群的平均存活率（48%）提高了69.35%。这种效应在100条鱼的外部种群中得到了验证。这些发现确定了与嗜水单胞杆菌耐药性相关的关键遗传标记，为阐明其潜在的分子免疫机制提供了方向，从而为未来的育种策略奠定了遗传基础。

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引用次数: 0

Sulforaphane attenuates HFD-induced enteritis in yellow river carp by restoring immune homeostasis, repairing intestinal barrier, and modulating gut microbiota 萝卜硫素通过恢复免疫稳态、修复肠道屏障和调节肠道菌群来减轻黄河鲤口蹄疫诱导的肠炎。

IF 3.9 2区农林科学 Q1 FISHERIES

Fish & shellfish immunology

Pub Date : 2026-01-19 DOI: 10.1016/j.fsi.2026.111125

Lu Wang , Bingke Wang , Jiaxiang Zhu , Qi Zhang , Yuxuan Cui , Huajuan Shi , Chunnuan Zhang

High-fat diets (HFD) exacerbate intestinal health risks in aquaculture. This study evaluated effects of dietary sulforaphane (SFN) supplementation on intestinal morphology, biochemistry, microbiota, and metabolism in Yellow River carp (Cyprinus carpio haematopterus) fed HFD. Five isoprotein diets diets were formulated: control (CN, 6 % lipid), HFD (12.29 % lipid), and HFD with 10 (HS10), 15 (HS15), or 20 (HS20) mg/kg SFN. Results demonstrated that HFD induced marked intestinal inflammation featuring villi deformation, oxidative stress, epithelial exfoliation, mitochondrial swelling, significantly elevated pro-inflammatory cytokines (TNF-α/IL-1β/IL-6; p < 0.05), and downregulated tight junction protein genes (Occludin/ZO-1/Claudin-3; p < 0.05), concomitant with microbial dysbiosis (Proteobacteria dominance>70 %, reduced Cetobacterium) and disrupted metabolic pathways (amino acid imbalance). SFN supplementation significantly enhanced antioxidant enzyme activities (SOD/CAT/GSH-Px, p < 0.05), peaking in HS15, which also exhibited minimized pro-inflammatory cytokines and maximized tight junction expression (p < 0.05). Furthermore, 16S rRNA gene sequencing of the gut microbiota indicated that SFN restored microbial homeostasis, notably by increasing the abundance of Cetobacterium. Metabolomic analysis based on KEGG enrichment revealed that these beneficial effects were associated with the activation of FoxO signaling, enhanced lysosomal function, and upregulation of coenzyme A biosynthesis. Critically, HS15 demonstrated optimal efficacy across all parameters. Therefore, 15 mg/kg SFN is identified as the optimal dose to ameliorate intestinal health in Yellow River carp, providing a theoretical basis for functional aquafeed development.

高脂肪饲料（HFD）加剧了水产养殖业的肠道健康风险。本研究评价了饲料中添加萝卜硫素（SFN）对饲喂高通量饲料的黄河鲤鱼（Cyprinus carpio haematopterus）肠道形态、生化、微生物群和代谢的影响。配制5种同蛋白饲料：对照组（CN， 6%脂质）、高脂饲料（12.29%脂质）和添加10 （HS10）、15 （HS15）和20 (HS20) mg/kg SFN的高脂饲料。结果表明，HFD诱导了明显的肠道炎症，表现为绒毛变形、氧化应激、上皮脱落、线粒体肿胀，促炎细胞因子（TNF-α/IL-1β/IL-6; p70%， Cetobacterium减少）显著升高，代谢途径中断（氨基酸失衡）。添加SFN显著提高了抗氧化酶活性(SOD/CAT/GSH-Px, p

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引用次数: 0

Dietary silymarin improves gut–liver health by enhancing intestinal barrier integrity and modulating TLR4/NF-κB and NLRP3-mediated inflammatory responses in Aeromonas hydrophila–infected grass carp 饲粮中添加水飞蓟素通过增强肠道屏障完整性和调节嗜水气单胞菌感染草鱼的TLR4/NF-κB和nlrp3介导的炎症反应来改善肠-肝健康。

IF 3.9 2区农林科学 Q1 FISHERIES

Fish & shellfish immunology

Pub Date : 2026-01-19 DOI: 10.1016/j.fsi.2026.111135

Feng Jia , Li Wei , Wei-Dan Jiang , Pei Wu , Yang Liu , Yao-Bin Ma , Hong-Yun Zhang , Xiao-Wan Jin , Hong-Mei Ren , Xiao-Qiu Zhou , Lin Feng

Bacterial infections compromise fish physiological homeostasis by impairing barrier function and metabolic organs, posing a persistent challenge in intensive aquaculture. Silymarin (SLM) is a plant-derived flavonoid complex obtained from Silybum marianum, widely recognized for its multifunctional pharmacological and bioactivity. However, its regulatory role under stress induced by bacterial pathogens in aquatic animals remains poorly understood. This study explored the impact and potential mechanisms of dietary SLM on intestinal barrier function and liver protection in juvenile grass carp. Fish (24.2 ± 0.1 g) were randomly assigned to six groups and fed diets containing 0, 20, 40, 60, 80, or 100 mg/kg SLM for 70 days. Subsequently, the fish were challenged with Aeromonas hydrophila (A. hydrophila) for 6 days. Optimal SLM supplementation (1) alleviated intestinal damage by upregulating tight junction proteins and downregulating myosin light chain kinase (MLCK); (2) reduced liver pro-inflammatory cytokines, inhibited lipopolysaccharide (LPS) translocation, and suppressed activation of the Toll-like receptor 4 (TLR4)/myeloid differentiation factor 88 (MyD88) signaling pathway; (3) mitigated liver inflammation by inhibiting the phosphorylation of key nuclear factor kappa B (NF-κB) and mitogen-activated protein kinase (MAPK) pathway proteins and downregulating nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3) inflammasome components; (4) decreased liver oxidative stress while enhancing antioxidant capacity and improving liver function; and (5) modulated gene expression related to metabolic enzymes and transporters in the intestine and liver. Dietary SLM improved gut–liver health by supporting barrier function, reducing inflammation, and enhancing metabolic and antioxidant responses, indicating its promise for nutritional use in aquaculture.

细菌感染通过损害屏障功能和代谢器官来破坏鱼类的生理稳态，对集约化水产养殖构成了持续的挑战。水飞蓟素（Silymarin， SLM）是从水飞蓟中提取的一种植物源性类黄酮复合物，因其具有多种药理和生物活性而被广泛认可。然而，它在水生动物细菌病原体诱导的应激下的调节作用仍然知之甚少。本研究旨在探讨饲料中添加SLM对草鱼幼鱼肠道屏障功能和肝脏保护的影响及其可能机制。试验鱼（24.2±0.1 g）随机分为6组，分别饲喂含0、20、40、60、80和100 mg/kg SLM的饲料，试验期70 d。随后，用嗜水气单胞菌攻毒6天。优化的SLM添加量(1)通过上调紧密连接蛋白和下调肌球蛋白轻链激酶（MLCK）来减轻肠道损伤；(2)降低肝脏促炎细胞因子，抑制脂多糖（LPS）易位，抑制toll样受体4 (TLR4)/髓样分化因子88 （MyD88）信号通路的激活；(3)通过抑制关键核因子κB （NF-κB）和丝裂原活化蛋白激酶（MAPK）途径蛋白的磷酸化和下调核苷酸结合寡聚结构域样受体蛋白3 （NLRP3）炎性小体成分，减轻肝脏炎症；(4)降低肝脏氧化应激，增强抗氧化能力，改善肝功能；(5)调节肠道和肝脏代谢酶和转运体相关基因的表达。饲粮SLM通过支持屏障功能、减少炎症、增强代谢和抗氧化反应来改善肠道-肝脏健康，表明其在水产养殖中的营养应用前景。

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引用次数: 0

CXCL20a promotes the immune protection of an inactivated vaccine against grass carp reovirus infection in grass carp (Ctenopharyngodon idella) CXCL20a促进草鱼呼肠孤病毒灭活疫苗对草鱼感染的免疫保护作用。

IF 3.9 2区农林科学 Q1 FISHERIES

Fish & shellfish immunology

Pub Date : 2026-01-19 DOI: 10.1016/j.fsi.2026.111140

Yizhong Lu , Shijie Wang , Yuezong Xu , Bo Tang , Guanyu Chen , Jianguo Su , Chunrong Yang

Grass carp (Ctenopharyngodon idella) is frequently threatened by hemorrhagic disease caused by grass carp reovirus (GCRV) infection in aquaculture. Vaccination represents a crucial strategy for preventing this infectious disease, where adjuvants play a pivotal role in enhancing vaccine efficacy. This study evaluated the chemokine CXCL20a as a molecular adjuvant to boost the immunogenicity of an inactivated GCRV vaccine. A CXCL20a fusion protein (rC20) was prepared and purified, and a tag-free form (Tf-rC20) was obtained after TrxA tag removal. Five experimental groups were established: PBS control, vaccine alone, rC20 alone, and vaccine combined with rC20 and Tf-rC20 in separate groups. The vaccine-alone group showed a 60 % survival rate, which increased to 74 % and 78 % in the Vac + rC20 and Vac + Tf-rC20 groups, respectively. Tf-rC20 showed slightly higher efficacy than rC20, although the difference was limited. Viral load quantification and splenic histopathology further confirmed that combined immunization suppressed viral replication and alleviated tissue damage. Serum immune parameters analyses revealed that combined immunization significantly enhanced the activities of lysozyme, total superoxide dismutase, and complement component C3, suggesting a potentiation of non-specific immunity. qRT-PCR results showed significant upregulation of immune-related genes, including IFN1, IL-6, Mx2, and IgM, in immune tissues, indicating that CXCL20a promoted both innate and adaptive immune responses. These results collectively demonstrate that the recombinant CXCL20a protein, functioning as a molecular adjuvant, can synergize with the inactivated vaccine to significantly enhance protective immunity against GCRV in grass carp, providing a promising candidate for the development of novel adjuvants in fish vaccines.

草鱼呼肠孤病毒（GCRV）感染引起的出血性疾病是养殖业中草鱼（Ctenopharyngodon idella）的常见威胁。疫苗接种是预防这种传染病的关键策略，其中佐剂在增强疫苗效力方面发挥关键作用。本研究评估了趋化因子CXCL20a作为分子佐剂增强灭活GCRV疫苗的免疫原性。制备并纯化了CXCL20a融合蛋白（rC20），去除TrxA标签后得到无标签形式（Tf-rC20）。实验组分为PBS对照组、单独接种疫苗组、单独接种rC20组、疫苗与rC20、Tf-rC20联合接种5组。单独接种疫苗组的存活率为60%，而Vac+rC20组和Vac+Tf-rC20组的存活率分别提高到74%和78%。Tf-rC20的疗效略高于rC20，但差异有限。病毒载量定量和脾组织病理学进一步证实联合免疫抑制病毒复制，减轻组织损伤。血清免疫参数分析显示，联合免疫显著提高了溶菌酶、总超氧化物歧化酶和补体成分C3的活性，提示非特异性免疫增强。qRT-PCR结果显示免疫组织中IFN1、IL-6、Mx2、IgM等免疫相关基因显著上调，表明CXCL20a促进了先天和适应性免疫反应。综上所示，重组CXCL20a蛋白作为一种分子佐剂，可与草鱼灭活疫苗协同作用，显著增强草鱼对GCRV的保护性免疫，为开发新型鱼疫苗佐剂提供了前景。

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引用次数: 0

Japanese eel SOCS1 suppresses MyD88-mediated NF-κB activation: implications for negative regulation of innate immunity in teleosts 日本鳗鱼SOCS1抑制myd88介导的NF-κB激活：对硬骨鱼先天免疫负调控的影响

IF 3.9 2区农林科学 Q1 FISHERIES

Fish & shellfish immunology

Pub Date : 2026-01-19 DOI: 10.1016/j.fsi.2026.111138

Tianyu Wang , Peng Lin , Yilei Wang , Luis O.B. Afonso , Jianjun Feng

Suppressor of cytokine signaling 1 (SOCS1) is a pivotal negative regulator of cytokine-mediated signaling in mammals, yet its role in teleost fish immunity remains poorly understood. In this study, we cloned and characterized the full-length SOCS1 cDNA from Japanese eel (Anguilla japonica), which encodes a 203-amino acid protein containing a conserved SH2 domain and SOCS box. Phylogenetic analysis placed AjSOCS1 within the teleost clade, showing high conservation with other vertebrate SOCS1 homologs. AjSOCS1 was ubiquitously expressed across tissues, with the highest levels in intestine and immune-related organs. Its transcription was significantly induced both in vivo (lipopolysaccharide [LPS], polyinosinic–polycytidylic acid [poly(I:C)], Aeromonas hydrophila) and in vitro (pathogen-associated molecular patterns [PAMPs] and bacterial infection), highlighting responsiveness to bacterial and viral stimuli. Subcellular localization analysis demonstrated that AjSOCS1 is cytoplasmic, consistent with its role in intracellular signaling regulation. Functional assays using a dual-luciferase reporter system revealed that AjSOCS1 strongly suppressed NF-κB activation under basal conditions, as well as during MyD88-, LPS-, and poly(I:C)-induced stimulation, and overexpression of AjSOCS1 resulted in coordinated downregulation of NF-κB subunits and downstream inflammatory and antimicrobial genes. Together, these findings provide the first functional evidence that SOCS1 in Japanese eel acts as a negative regulator of NF-κB signaling, thereby extending the known repertoire of SOCS1 activity in fish. This study not only enriches comparative immunology of SOCS proteins but also has potential implications for controlling excessive inflammation and improving disease resistance in aquaculture.

细胞因子信号传导抑制因子1 （SOCS1）是哺乳动物细胞因子介导的信号传导的关键负调节因子，但其在硬骨鱼免疫中的作用尚不清楚。本研究克隆并鉴定了日本鳗鲡（Anguilla japonica）全长SOCS1 cDNA，该基因编码一个含有保守SH2结构域和SOCS盒的203个氨基酸的蛋白。系统发育分析表明，AjSOCS1属于硬骨鱼分支，与其他脊椎动物SOCS1同源物具有高度的保守性。AjSOCS1在各组织中普遍表达，在肠道和免疫相关器官中表达水平最高。它的转录在体内（脂多糖[LPS]、多肌苷-多胞酸[poly(I:C)]、嗜水气单胞菌）和体外（病原体相关分子模式[PAMPs]和细菌感染）均被显著诱导，突出了对细菌和病毒刺激的反应性。亚细胞定位分析表明，AjSOCS1是细胞质的，与其在细胞内信号调节中的作用一致。使用双荧光素酶报告系统进行的功能分析显示，AjSOCS1在基础条件下以及MyD88-、LPS-和poly(I:C)-诱导的刺激下强烈抑制NF-κ b的激活，并且AjSOCS1的过表达导致NF-κ b亚基和下游炎症和抗菌基因的协同下调。总之，这些发现提供了第一个功能性证据，证明日本鳗鱼中的SOCS1作为NF-κB信号的负调节因子，从而扩展了鱼类中SOCS1活性的已知曲目。本研究不仅丰富了SOCS蛋白的比较免疫学，而且对控制水产养殖过度炎症和提高抗病性具有潜在的指导意义。

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引用次数: 0

RBX1 and RBX2 promote GCRV replication in grass carp (Ctenopharyngodon idella) RBX1和RBX2促进草鱼GCRV的复制。

IF 3.9 2区农林科学 Q1 FISHERIES

Fish & shellfish immunology

Pub Date : 2026-01-19 DOI: 10.1016/j.fsi.2026.111142

Jianhua Feng , Yafang Wang , Wenji Huang , Dawei Xie , Yinghao Huang , Xinran Tao , Tiantian Zhang , Haixia Wu , Jun Zou , Junya Wang

Cullin-RING ligases (CRLs) constitute the most diverse E3 ligase family in the ubiquitin-proteasome system, yet their antiviral roles in bony fish remain poorly understood. Here, we identify the catalytic cores of CRLs, the RING-box proteins RBX1 and RBX2, as key modulators of antiviral signaling in cyprinid fish. Phylogenetic and structural analyses revealed that Rbx1 and Rbx2 are highly conserved and widely expressed, with preferential enrichment in primordial germ and immune-related cells. Upon infection with grass carp reovirus (GCRV) or spring viremia of carp virus (SVCV) in cyprinid fish, Rbxs were rapidly induced in vivo and in vitro. RBX1 and RBX2 synergistically promote viral replication by directly interacting with interferon regulatory factors (IRF) 3 and IRF7 and facilitating their ubiquitin-mediated regulation. Together, our findings uncover RBX1 and RBX2 as evolutionarily conserved negative regulators of fish innate immunity and provide mechanistic insight into the ubiquitin-mediated control of interferon homeostasis across vertebrates.

Cullin-RING连接酶（CRLs）构成了泛素-蛋白酶体系统中最多样化的E3连接酶家族，但它们在硬骨鱼中的抗病毒作用仍然知之甚少。在这里，我们确定了crl的催化核心，RING-box蛋白RBX1和RBX2，作为鲤科鱼类抗病毒信号的关键调节剂。系统发育和结构分析表明，Rbx1和Rbx2是高度保守和广泛表达的，在原始生殖和免疫相关细胞中优先富集。用草鱼呼肠孤病毒（GCRV）或鲤科鱼春季病毒血症（SVCV）感染鲤科鱼后，在体内和体外快速诱导Rbxs。RBX1和RBX2通过直接与干扰素调节因子（IRF） 3和IRF7相互作用，促进其泛素介导的调控，协同促进病毒复制。总之，我们的研究结果揭示了RBX1和RBX2是进化上保守的鱼类先天免疫负调节因子，并为泛素介导的干扰素稳态控制在脊椎动物中的机制提供了见解。

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引用次数: 0