Fish & shellfish immunology最新文献_第5页

Evolution and expression of TLR5a and TLR5b in lamprey (Lampetra japonica) TLR5a和TLR5b在七鳃鳗中的进化与表达。

IF 4.1 2区农林科学 Q1 FISHERIES

Fish & shellfish immunology

Pub Date : 2025-02-01 DOI: 10.1016/j.fsi.2024.110061

Lutian Wang , Fan Hui , Yu Liu , Yijie Zhang , Weiyin Xu , Qingkui Bai , Ting Zhu

The lamprey serves as a key model organism for studying the origin and evolution of species, embryonic development, and the immune system. The immune system primarily relies on pattern recognition receptors, including Toll-like receptors (TLRs), with Toll-like receptor 5 (TLR5) having a particularly complex evolutionary history. Currently, although TLR5 is being identified in an expanding array of taxonomic groups, a comprehensive study on its evolutionary aspects is yet to be conducted. In this study, we identified Lj-TLR5a and Lj-TLR5b in Lampetra japonica and examined their distribution across various tissues in this species. Furthermore, we conducted preliminary investigations into their immune functions and discovered that, as primitive genes, they are highly sensitive to various pathogens. Upon recognizing flagellar proteins, both Lj-TLR5a and Lj-TLR5b work together; however, these TLRs may function independently in response to other stimuli. Subsequently, we performed comprehensive structural and evolutionary analyses of the TLR5 family, incorporating TLR5 data from various species at different evolutionary stages. Our findings revealed that TLR5a and TLR5b in lamprey are in a relatively primitive evolutionary state. Meanwhile, TLR5L differentiated during the early stages of evolution and exhibits a pseudogenic trend throughout this evolutionary process; notably, this TLR is currently preserved only in certain amphibian and reptile species. In cartilaginous fish, only one type of TLR5 is usually retained, whereas bony fish typically possess both TLR5 and TLR5S. TLR5S has a relatively simple structure, likely arising from repetitive whole-genome events in bony fishes. Among bony fishes, flesh-finned fish were found to retain only one TLR5, which eventually evolved into the TLR5 found in quadrupeds. In summary, this study provides significant insights into the origin and evolution of the TLR5 family by analyzing the evolutionary status and immune functions of Lj-TLR5a and Lj-TLR5b in Japanese lampreys.

七鳃鳗是研究物种起源和进化、胚胎发育和免疫系统的关键模式生物。免疫系统主要依赖于模式识别受体，包括toll样受体（TLRs），其中toll样受体5 （TLR5）具有特别复杂的进化史。目前，虽然在越来越多的分类群中发现了TLR5，但对其进化方面的全面研究尚未开展。本研究在日本七鳃鳗中鉴定出Lj-TLR5a和Lj-TLR5b，并研究了它们在该物种各组织中的分布。此外，我们对其免疫功能进行了初步调查，发现作为原始基因，它们对各种病原体高度敏感。在识别鞭毛蛋白时，Lj-TLR5a和Lj-TLR5b协同工作；然而，这些tlr可能在对其他刺激的反应中独立起作用。随后，我们对TLR5家族进行了全面的结构和进化分析，纳入了不同物种在不同进化阶段的TLR5数据。结果表明，TLR5a和TLR5b在七鳃鳗中处于相对原始的进化状态。与此同时，TLR5L在进化早期就已分化，并在整个进化过程中呈现假基因倾向；值得注意的是，这种TLR目前只保存在某些两栖动物和爬行动物物种中。在软骨鱼中，通常只保留一种TLR5，而硬骨鱼通常同时拥有TLR5和TLR5S。TLR5S结构相对简单，可能源于硬骨鱼类重复的全基因组事件。在硬骨鱼类中，肉鳍鱼被发现只保留了一个TLR5，最终进化成四足动物的TLR5。综上所述，本研究通过分析Lj-TLR5a和Lj-TLR5b在日本七鳃鳗中的进化状态和免疫功能，对TLR5家族的起源和进化有重要意义。

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引用次数: 0

Roles of S100A1 and S100A10 from hybrid grouper (Epinephelus lanceolatus♂ × Epinephelus fuscoguttatus♀) in immune response to Vibrio infection 杂交石斑鱼（Epinephelus lanceolatus♂× Epinephelus fuscoguttatus♀）S100A1和S100A10在弧菌感染免疫应答中的作用

IF 4.1 2区农林科学 Q1 FISHERIES

Fish & shellfish immunology

Pub Date : 2025-02-01 DOI: 10.1016/j.fsi.2024.110070

Sheng Yuan , Ying Huang , Quan Wang , Yan Shi , Xiao-Rui Song , Zhe Zhao

The S100 proteins are highly conserved EF-hand calcium-binding proteins found only in vertebrates. In the current study, two S100 genes (S100A1 and S100A10) were successfully identified and characterized from hybrid grouper Epinephelus lanceolatus♂ × Epinephelus fuscoguttatus♀. The deduced S100A10 protein contained two EF-hand domains, and S100A1 only possessed the N-terminal EF-hand. Phylogenetic analysis revealed that S100A1 and S100A10 from hybrid grouper were evolutionarily closely related to their counterparts in other selected vertebrates. Quantitative real-time PCR results revealed that the transcripts of S100A1 and S100A10 mRNA were ubiquitously distributed in all the examined tissues. After Vibrio alginolyticus infection, the expression of S100A1 and S100A10 in the spleen increased significantly. Moreover, overexpression of S100A1 and S100A10 could not only regulate the expression of interleukin 8 (IL-8), IL-10, and IL-16 in the head kidney, liver, and spleen, change the activities of acid phosphatase, catalase, lysozyme, and superoxide dismutase in serum, but also reduce the promoter activities of interferon 3 and nuclear factor kappa-B in vitro. Taken together, this study indicated that S100A1 and S100A10 participate in the immune response of hybrid grouper against bacterial infection.

S100蛋白是高度保守的EF-hand钙结合蛋白，仅存在于脊椎动物中。本研究从杂交石斑鱼Epinephelus lanceolatus♂× Epinephelus fuscoguttatus♀中成功鉴定并鉴定了S100基因S100A1和S100A10。推导出的S100A10蛋白含有两个EF-hand结构域，而S100A1仅具有n端EF-hand结构域。系统发育分析表明，杂交石斑鱼的S100A1和S100A10与其他脊椎动物的S100A1和S100A10在进化上具有密切的亲缘关系。实时荧光定量PCR结果显示，S100A1和S100A10 mRNA转录本在所有检测组织中普遍存在。感染溶藻弧菌后，脾脏中S100A1和S100A10的表达显著升高。此外，过表达S100A1和S100A10不仅可以调节头肾、肝、脾组织中白细胞介素8 （IL-8）、IL-10和IL-16的表达，改变血清酸性磷酸酶、过氧化氢酶、溶菌酶和超氧化物歧化酶的活性，还可以降低体外干扰素3和核因子κ b的启动子活性。综上所述，本研究提示S100A1和S100A10参与了杂交石斑鱼对细菌感染的免疫应答。

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引用次数: 0

White spot syndrome virus IE1 protein blocks the integrin-FAK signaling to enhance viral infection in shrimp 白斑综合征病毒IE1蛋白阻断整合素- fak信号通路增强对虾病毒感染。

IF 4.1 2区农林科学 Q1 FISHERIES

Fish & shellfish immunology

Pub Date : 2025-02-01 DOI: 10.1016/j.fsi.2024.110073

Kaiyu Lu , Xiyu Chen , Huimin Zhang , Jinghua Zhu , Yongzhen Zhao , Xiuli Chen , Yueling Zhang , Defu Yao

DNA viruses commonly utilize immediate-early proteins to manipulate cellular signaling pathways in order to facilitate their infection. Our previous research has suggested that IE1, an immediate-early protein encoded by the white spot syndrome virus (WSSV), may modulate the shrimp integrin-FAK signaling pathway. However, the specific molecular mechanism and role of IE1 in regulating this signaling pathway remain unclear. In this study, we demonstrated that IE1 competes for binding to the cytoplasmic tail of Penaeus vannamei integrin-α5, resulting in the inhibition of the integrin-α5-FAK interaction, thereby suppressing FAK activation and cell adhesion. Furthermore, we observed a significant increase in the expression of P. vannamei integrin-α5 and FAK following WSSV infection. Additionally, knockdown of integrin-α5 or FAK through RNA interference has been shown to reduce cell adhesion and enhance WSSV infection. In conclusion, our findings reveal that IE1 disrupts integrin-FAK signaling to inhibit cell adhesion, ultimately promoting WSSV infection in shrimp.

DNA病毒通常利用即时早期蛋白来操纵细胞信号通路，以促进其感染。我们之前的研究表明，白斑综合征病毒（WSSV）编码的即早蛋白IE1可能调控虾整合素- fak信号通路。然而，IE1在调控这一信号通路中的具体分子机制和作用尚不清楚。在本研究中，我们证明了IE1与南美对虾整合素-α5的细胞质尾部竞争结合，从而抑制整合素-α5-FAK的相互作用，从而抑制FAK的活化和细胞粘附。此外，我们观察到在WSSV感染后，凡纳梅整合素-α5和FAK的表达显著增加。此外，通过RNA干扰敲低整合素-α5或FAK可降低细胞粘附并增强WSSV感染。总之，我们的研究结果表明，IE1破坏整合素- fak信号以抑制细胞粘附，最终促进虾的WSSV感染。

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引用次数: 0

Unravelling the ontogeny and tissue-specific expression profiles of immune-related genes in the near-threatened endemic catfish, Clarias dussumieri 揭示近危地方性鲶鱼的个体发生和免疫相关基因的组织特异性表达谱。

IF 4.1 2区农林科学 Q1 FISHERIES

Fish & shellfish immunology

Pub Date : 2025-02-01 DOI: 10.1016/j.fsi.2024.110075

Sagar Gorakh Satkar , Arun Sudhagar , Arathi Dharmaratnam , Thangaraj Raja Swaminathan , Neeraj Sood , Cheru Parambil Abhilash , Ravi Charan , Uttam Kumar Sarkar

Clarias dussumieri, an air breathing catfish endemic to the Western Ghats in India, is categorized as ‘Near Threatened’ by the IUCN. This species is of high regional consumer demand and is one of the prioritized candidate species for aquaculture diversification and conservation. Despite its ecological and commercial significance, comprehensive studies on its immune system are lacking. This study elucidates the ontogenetic development and tissue-specific (brain, anterior kidney, gill, spleen, muscle, liver, hindgut and skin) expression profiles of key immune-related genes in C. dussumieri. Larvae were sampled at various developmental stages post-fertilization, and tissues from adult fish were analyzed for expression patterns of genes associated with inflammation (IL-1β, TNF-α, iNOS), antimicrobial defense (LYS, HAMP), stress response (HSP70), complement system (C3), cell-mediated immunity (MHC-IIβ, CD4-1), and adaptive immunity (IgM). The expression of IL-1β was highest at 60 days post-fertilization (60D), while TNF-α expression peaked at 25D before dropping notably by 60D. iNOS showed a peak at 7D, underscoring its importance in early immune defence. LYS exhibited a high expression at 10D, while HAMP peaked at 60D, highlighting their roles in antimicrobial defence. Stress marker HSP70 increased from 15D onwards and complement component C3 was consistently expressed at low levels throughout development. MHC-IIβ and CD4-1 showed significant increase since 10D and 7D respectively, suggesting the establishment of cell-mediated immunity. IgM expression increased notably from 15 days, indicating the development of adaptive immunity. In adult fish tissues, IL-1β, TNF-α, LYS and HSP70 showed highest expression in the hindgut, while C3 was predominantly expressed in the liver. MHC-IIβ and CD4-1 were highly expressed in the spleen and anterior kidney respectively. IgM was abundant in the anterior kidney and spleen. This study provides crucial baseline data on the immune competence of different developmental stages of C. dussumieri, informing strategies for effective vaccination and disease management in aquaculture, and enhancing our understanding of fish immunology for conservation efforts.

Clarias dussumieri是印度西高止山脉特有的一种呼吸空气的鲶鱼，被世界自然保护联盟列为“近危”物种。该物种具有较高的区域消费需求，是水产养殖多样化和保护的优先候选物种之一。尽管其具有生态和商业意义，但缺乏对其免疫系统的全面研究。本研究阐明了dusumieri的个体发育和组织特异性（脑、前肾、鳃、脾脏、肌肉、肝脏、后肠和皮肤）关键免疫相关基因的表达谱。在受精后的不同发育阶段采集幼虫，分析成鱼组织中与炎症（IL-1β、TNF-α、iNOS）、抗菌防御（LYS、HAMP）、应激反应（HSP70）、补体系统（C3）、细胞介导免疫（MHC-IIβ、CD4-1）和适应性免疫（IgM）相关基因的表达模式。IL-1β在受精后60天（60D）表达最高，TNF-α在受精后25D表达最高，60D后显著下降。iNOS在7D时达到峰值，强调了其在早期免疫防御中的重要性。LYS在10D时高表达，而HAMP在60D时达到峰值，突出了它们在抗菌防御中的作用。应激标记HSP70从15D开始增加，补体成分C3在整个发育过程中一直以低水平表达。从10D和7D开始，mhc - i β和CD4-1分别显著升高，提示细胞介导免疫的建立。从第15天开始，IgM表达显著增加，表明适应性免疫的发展。在成鱼组织中，IL-1β、TNF-α、LYS和HSP70在后肠中表达最高，而C3主要在肝脏中表达。mhc - i β和CD4-1分别在脾脏和前肾高表达。IgM在前肾和脾中含量丰富。该研究为不同发育阶段的杜氏梭菌免疫能力提供了重要的基线数据，为水产养殖中有效的疫苗接种和疾病管理策略提供了信息，并增强了我们对鱼类免疫学的了解，为保护工作提供了依据。

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引用次数: 0

WSSV induces Rubicon expression to regulate innate immune response in Penaeus vannamei WSSV 可诱导 Rubicon 的表达，从而调节万年青中的先天免疫反应。

IF 4.1 2区农林科学 Q1 FISHERIES

Fish & shellfish immunology

Pub Date : 2025-02-01 DOI: 10.1016/j.fsi.2024.110033

Phattarunda Jaree, Thaneeya Nantapojd, Chalermporn Ongvarrasopone

Rubicon, the RUN domain Beclin-1-interacting and cysteine-rich domain-containing protein plays an important role in facilitating viral replication. In this study, an involvement of P. vannamei Rubicon or PvRUBCN during white spot syndrome virus (WSSV) infection and its roles in regulation of apoptosis and innate immune response were investigated. The full-length coding sequence of PvRUBCN was 3708 bp encoding the protein of 1235 amino acids. PvRUBCN contained three conserved domains including the RUN, the PI3K-binding, and the Rubicon homology domains. PvRUBCN was grouped with the closely related RUBCN from the Penaeid species with more than 95 % identity. It was highly expressed in nerve followed by intestine, and gill. Its expression was induced upon WSSV challenge at 12 and 48 hpi. Suppression of PvRUBCN by dsRNA upon WSSV challenge resulted in more than 80 % reduction in PvRUBCN mRNA expression. Knockdown of PvRUBCN mRNA significantly decreased the WSSV copy number and prolonged the survival rate of WSSV-infected shrimp. In addition, the caspase3, a key regulator of the apoptosis pathway was significantly down-regulated. The interferon like genes including Vago4 was dramatically decreased at 72 hpi whereas Vago5 demonstrated slight reduction at 24 hpi and increase at 72 and 120 hpi, respectively. On the other hand, the expressions of the genes involved in the prophenoloxidase pathway including PPO1 and PPO2 were not changed. Taken together, PvRUBCN played important roles in the antiviral immunity in response to WSSV infection.

Rubicon是RUN结构域Beclin-1-interacting和富含半胱氨酸结构域的蛋白，在促进病毒复制方面发挥着重要作用。本研究探讨了凡纳米星虫Rubicon或PvRUBCN在白斑综合征病毒（WSSV）感染过程中的参与及其在调控细胞凋亡和先天性免疫反应中的作用。PvRUBCN的全长编码序列为3,708 bp，编码1,235个氨基酸的蛋白质。PvRUBCN 包含三个保守结构域，包括 RUN、PI3K 结合和 Rubicon 同源结构域。PvRUBCN 与青鳉鱼类中密切相关的 RUBCN 同源度超过 95%。它在神经中高表达，其次是肠和鳃。在 12 hpi 和 48 hpi 受到 WSSV 挑战时，PvRUBCN 会被诱导表达。在WSSV挑战下，通过dsRNA抑制PvRUBCN会导致PvRUBCN mRNA表达量减少80%以上。PvRUBCN mRNA的敲除能显著降低WSSV拷贝数并延长WSSV感染虾的存活率。此外，凋亡途径的关键调控因子 Caspase3 的表达也明显下调。包括 Vago4 在内的干扰素样基因在 72 hpi 时急剧下降，而 Vago5 则在 24 hpi 时略有下降，在 72 hpi 和 120 hpi 时分别有所上升。另一方面，参与丙酚氧化酶通路的基因（包括 PPO1 和 PPO2）的表达没有变化。综上所述，PvRUBCN在应对WSSV感染的抗病毒免疫中发挥了重要作用。

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引用次数: 0

CRISPR/Cas9-induced knockout of tumor necrosis factor-alpha-type I augments viral infection in zebrafish CRISPR/ cas9诱导的肿瘤坏死因子α - I型敲除增加斑马鱼的病毒感染。

IF 4.1 2区农林科学 Q1 FISHERIES

Fish & shellfish immunology

Pub Date : 2025-02-01 DOI: 10.1016/j.fsi.2024.110092

Arthika Kalaichelvan , Kishanthini Nadarajapillai , Sarithaa Raguvaran Sellaththurai , U.P.E. Arachchi , Myoung-Jin Kim , Sumi Jung , Jehee Lee

Tumor necrosis factor-alpha (TNF-α) is a pleiotropic cytokine with critical roles in inflammation, cell survival, and defense. As a member of the TNF superfamily, it exerts its effects by binding to transmembrane receptors and triggering various downstream signaling pathways. Although TNF-α′s involvement in antiviral responses in mammals is well-established, its role in teleost remains poorly understood. This study investigated the contribution of TNF-α1 to antiviral immunity in zebrafish using a tnf-α1^(−/−) knockout (KO) line. We challenged both wild-type and tnf-α1^(−/−) zebrafish with viral hemorrhagic septicemia virus (VHSV) at both embryonic and adult stages. Mortality was observed at 4 days post-infection (dpi) in tnf-α1-deficient adult fish challenged with 5 × 10⁶ TCID₅₀ (VHSV) and at 5 dpi in adult wild fish challenged with the same concentration. In addition, tnf-α1^(−/−) KO adult fish reached the maximum mortality of 100 % at 20 dpi, whereas wild adult fish reached 54 % mortality at the same time point. This increased susceptibility to early mortality was associated with a higher viral burden and altered expression of key immune genes, including the pro-inflammatory cytokines il-6 and il-1β, the anti-inflammatory cytokine il-10, and interferon-related genes such as irf1 and ifn-γ. Our findings demonstrate the crucial role of TNF-α1 in antiviral defense mechanisms in zebrafish and provide valuable insights into the functional conservation of TNF-α signaling across vertebrate species. This knowledge may contribute to the development of strategies to combat viral diseases in fish.

肿瘤坏死因子-α (TNF-α)是一种多效性细胞因子，在炎症、细胞存活和防御中起重要作用。作为TNF超家族的一员，它通过与跨膜受体结合，触发多种下游信号通路发挥作用。虽然TNF-α参与哺乳动物的抗病毒反应是公认的，但其在硬骨鱼中的作用仍然知之甚少。本研究利用TNF-α1（-/-）敲除（KO）细胞系研究了TNF-α1对斑马鱼抗病毒免疫的贡献。我们用病毒出血性败血症病毒（VHSV）攻击野生型和tnf-α1（-/-）斑马鱼的胚胎期和成虫期。用5×106 TCID50 （VHSV）侵染tnf-α1缺陷成鱼，感染后4天死亡，用相同浓度侵染野生成鱼感染后5天死亡。此外，tnf-α1(-/-) KO成鱼在20 dpi时最高死亡率为100%，而野生成鱼在同一时间点的死亡率为54%。这种对早期死亡的易感性增加与较高的病毒负荷和关键免疫基因的表达改变有关，包括促炎细胞因子il-6和il-1β、抗炎细胞因子il-10和干扰素相关基因如irf1和ifn-γ。我们的研究结果证明了TNF-α1在斑马鱼抗病毒防御机制中的关键作用，并为TNF-α信号在脊椎动物物种中的功能保护提供了有价值的见解。这一知识可能有助于制定对抗鱼类病毒性疾病的策略。

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引用次数: 0

A vaccine combining ORF132 and ORF25 expressed by Saccharomyces cerevisiae induces protective immunity in Carassius auratus gibelio against CyHV-2 一种由酿酒酵母表达的ORF132和ORF25结合的疫苗可诱导异育银鲫对CyHV-2的保护性免疫。

IF 4.1 2区农林科学 Q1 FISHERIES

Fish & shellfish immunology

Pub Date : 2025-02-01 DOI: 10.1016/j.fsi.2024.110099

Jiahui Liang , Maoxia Yang , Xinmei Li , Qianjin Zhou , Guanjun Yang , Jianfei Lu , Jiong Chen

CyHV-2 is the pathogen of herpesvirus hematopoietic necrosis (HVHN), resulting in significant economic losses in crucian carp. Although multiple oral vaccines have been developed to prevent CyHV-2, they have not achieved ideal protective effects. To improve the protective effect of oral vaccine, a combination vaccine was conducted by mixing recombinant Saccharomyces cerevisiae displaying ORF132 or ORF25 on the cell surface in a 1:1 ratio. Oral immunization with this combined vaccine induced specific antibodies against ORF132 and ORF25 in fish serum, as well as increased the transcription levels of IgT, IgM, IL-1β, IFN-1, and TNF-α in the spleen, head-kidney, and hindgut. Importantly, the combined vaccine provided significant protection against CyHV-2 infection in crucian carp, resulting in a relative percent survival of 80.77 %. Additionally, it suppressed viral load and alleviated pathological damage of CyHV-2-infected fish. The results suggest that the combined vaccine exhibits a better protective effect compared to the single-antigen vaccines, indicating it as a promising approach to prevent CyHV-2 outbreaks.

CyHV-2是疱疹病毒造血坏死（HVHN）的病原体，给鲫鱼造成重大的经济损失。虽然已经开发出多种口服疫苗来预防CyHV-2，但它们并没有达到理想的保护效果。为了提高口服疫苗的保护作用，将重组酵母（ORF132或ORF25）按1:1的比例混合在细胞表面，制备了一种联合疫苗。口服免疫该联合疫苗可诱导鱼血清中针对ORF132和ORF25的特异性抗体，并增加脾脏、头肾和后肠中IgT、IgM、IL-1β、IFN-1和TNF-α的转录水平。重要的是，联合疫苗对鲫鱼CyHV-2感染具有显著的保护作用，相对存活率为80.77%。此外，它还能抑制病毒载量，减轻cyhv -2感染鱼的病理损伤。结果表明，与单抗原疫苗相比，联合疫苗表现出更好的保护效果，表明它是预防CyHV-2爆发的一种有希望的方法。

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引用次数: 0

Identification of the arachidonic acid 5-lipoxygenase and its function in the immunity of Apostichopus japonicus 花生四烯酸5-脂氧合酶的鉴定及其在刺参免疫中的作用。

IF 4.1 2区农林科学 Q1 FISHERIES

Fish & shellfish immunology

Pub Date : 2025-02-01 DOI: 10.1016/j.fsi.2024.110095

Jiahao Cheng , Kai Yang , Xu Li , Benkang Liu , Ming Chen , Cheng Li

A number of studies have been demonstrated that arachidonate 5-lipoxygenase (ALOX-5) plays a role in regulating a range of physiological and pathological processes through the catalysis of leukotriene formation from arachidonic acid (ARA). The coding sequence of ALOX-5 from Apostichopus japonicus (Aj-ALOX-5) was successfully amplified, resulting in a 2028 bp ORF sequence that encodes 674 amino acids. A comparison of the amino acid sequence with those of other 5-lipoxygenases revealed that Aj-ALOX-5 has the N-terminal "PLAT domain" and C-terminal "lipoxygenase structural domain" characteristic of this enzyme family. The enzyme activity sites and Ca²⁺-binding sites exhibited high levels of conservation. The phylogenetic tree also indicated that Aj-ALOX-5 was closely related to starfish 5-lipoxygenase. The recombinant Aj-ALOX-5 (rAj-ALOX-5) was obtained through the exogenous expression of an engineered bacterium and purified using Ni²⁺-NTA. rAj-ALOX-5 was observed to catalyze ARA to produce 5-HPETE and LTA₄, which indicated that the Aj-ALOX-5 protein belonged to the 5-lipoxygenase family. qRT-PCR demonstrated that Aj-ALOX-5 is widely distributed in tissues. Furthermore, the Aj-ALOX-5 mRNA and the production of 5-HETE were found to be significantly up-regulated in response to stress induced by Vibrio splendidus. Inhibition of Aj-ALOX-5 expression by the optimal caffeic acid resulted in a significant increase in mortality rates of sea cucumbers. Further investigation revealed that the production of 5-HPETE and NF-κB I was also significantly suppressed. It can be hypothesized that Aj-ALOX-5 plays an important role in the immune response of sea cucumbers by mediating the NF-κB I pathway.

大量研究表明，花生四烯酸5-脂氧合酶（ALOX-5）通过催化花生四烯酸（ARA）生成白三烯，在调节一系列生理和病理过程中起作用。成功扩增出Apostichopus japonicus ALOX-5的编码序列（Aj-ALOX-5），得到2028 bp的ORF序列，编码674个氨基酸。与其他5-脂氧合酶的氨基酸序列比较发现，Aj-ALOX-5具有该酶家族的n端“PLAT结构域”和c端“脂氧合酶结构域”特征。酶活性位点和Ca2 +结合位点表现出高度的保守性。系统进化树还表明，Aj-ALOX-5与海星5-脂氧合酶亲缘关系密切。重组Aj-ALOX-5 （rAj-ALOX-5）通过工程菌外源表达获得，用Ni2+-NTA纯化。观察到rAj-ALOX-5能催化ARA生成5-HPETE和LTA4，表明Aj-ALOX-5蛋白属于5-脂氧合酶家族。qRT-PCR结果显示Aj-ALOX-5广泛分布于组织中。此外，在应激条件下，脾脏弧菌诱导的Aj-ALOX-5 mRNA和5-HETE的表达均显著上调。最佳咖啡酸对Aj-ALOX-5表达的抑制显著提高了海参的死亡率。进一步研究发现，5-HPETE和NF-κB I的产生也被显著抑制。可以推测Aj-ALOX-5通过介导NF-κB I通路在海参免疫应答中发挥重要作用。

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引用次数: 0

Impact of Bambusa vulgaris-supplemented diet on Nile tilapia challenged with Pseudomonas putida: Hematological, immune, and oxidative responses 添加竹节饲料对感染恶臭假单胞菌尼罗罗非鱼的影响：血液学、免疫和氧化反应。

IF 4.1 2区农林科学 Q1 FISHERIES

Fish & shellfish immunology

Pub Date : 2025-02-01 DOI: 10.1016/j.fsi.2024.110102

Alamira Marzouk Fouad , Haitham G. Abo-Al-Ela , Rehab H. Moneeb , Mariana S. Alfons , Abdallah S. Salah , Shaymaa Yusuf

This study investigated the effects of bamboo shoot extract (Bambusa vulgaris) as a feed additive on the health profiles and infection resistance of Nile tilapia (Oreochromis niloticus) against Pseudomonas putida. Bamboo shoot extract was added at levels of 0 g, 40 g, and 60 g per 1000 g of diet over a 60-day period. The fish were then challenged with a pathogenic P. putida strain. Chemical analysis of the bamboo shoot extract identified 3,5-dinitrophenol and hydroquinone as the two most abundant compounds. Results showed that fish fed bamboo-enriched diets exhibited significantly enhanced levels of red blood cells, hemoglobin, hematocrit, white blood cells, and platelets, and improved erythrocyte cellular and nuclear morphologies, indicating improved health profiles after the challenge. Liver function indicators, including AST, ALT, and ALP, were notably balanced in fish receiving bamboo shoot extract post-challenge (p < 0.05). Blood levels of K⁺ were lower in the bamboo-fed groups. Additionally, blood levels of Ca⁺⁺ and Na⁺ were reduced in fish fed 40 g and 60 g of bamboo, respectively, compared to the control group (p < 0.01). The bamboo extract also enhanced immune and oxidative capacities, as demonstrated by increased catalase, superoxide dismutase, lysozyme activity, and phagocytic activity, along with reduced malondialdehyde levels and elevated serum immunoglobulin M (p < 0.01). Gene expression analysis revealed significant effects of Bambusa vulgaris extract, Pseudomonas infection, and their interaction on the expression of interleukin-1β, interleukin-10, and NK-lysin genes, with varying expression levels at 1, 3, and 7 days post-challenge (p < 0.05). The liver bacterial load in fish exposed to P. putida significantly decreased in the bamboo-fed groups, with the lowest count observed in the 60 g bamboo group. Additionally, survival rates were markedly higher in the bamboo-fed groups compared to the control, with no significant difference between the two bamboo-fed groups. In conclusion, dietary supplementation with bamboo shoot extract enhances hematological parameters, blood cell and nuclear morphology, and increases survival rates in Nile tilapia following infection.

本试验研究了竹笋提取物作为饲料添加剂对尼罗罗非鱼（Oreochromis niloticus）健康状况和对恶臭假单孢菌的抗性的影响。在60天的试验期内，每1000 g日粮中分别添加0 g、40 g和60 g竹笋提取物。然后用致病性恶臭假单胞菌菌株攻毒鱼。竹笋提取物的化学分析鉴定出3,5-二硝基苯酚和对苯二酚是两种含量最高的化合物。结果表明，喂食富含竹子的饲料后，鱼的红细胞、血红蛋白、红细胞比压、白细胞和血小板水平显著提高，红细胞和细胞核形态也有所改善，表明鱼的健康状况得到改善。竹笋提取物攻毒后，鱼的肝功能指标AST、ALT和ALP均达到了显著平衡（p < 0.05）。以竹子为食的实验组血液中的钾离子水平较低。此外，饲喂40 g和60 g竹子的鱼血液中Ca++和Na+水平也分别低于对照组（p < 0.01）。竹提取物还能增强免疫和氧化能力，提高过氧化氢酶、超氧化物歧化酶、溶菌酶活性和吞噬活性，降低丙二醛水平，提高血清免疫球蛋白M （p < 0.01）。基因表达分析显示，竹节提取物、假单胞菌感染及其互作对攻毒后1、3、7 d白细胞介素-1β、白细胞介素-10和NK-lysin基因表达有显著影响（p < 0.05）。竹饲组暴露于恶臭假单胞菌的鱼的肝脏细菌负荷显著降低，其中以60 g竹饲组最低。此外，竹食组的存活率明显高于对照组，两组之间无显著差异。由此可见，饲粮中添加竹笋提取物可改善尼罗罗非鱼感染后的血液学指标、血细胞和细胞核形态，提高其存活率。

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引用次数: 0

The haemocyte highly-expressed E-type prostanoid receptor regulates TNF expression during immune response of oyster Crassostrea gigas 白细胞高表达的e型前列腺素受体调节牡蛎长牡蛎免疫应答过程中TNF的表达。

IF 4.1 2区农林科学 Q1 FISHERIES

Fish & shellfish immunology

Pub Date : 2025-02-01 DOI: 10.1016/j.fsi.2024.110108

Xuemei Cheng , Weilin Wang , Miren Dong , Junlei Cheng , Jiajun Zuo , Xiaoxu Zhou , Lingling Wang , Linsheng Song

Prostaglandin E2 imparts diverse physiological effects on multiple cells through its actions on four distinct E-type prostanoid (EP) receptor subtypes (EP1-EP4), among which the EP4 is one of subtypes known to mediate the immune response in mammalian monocytes and macrophages. However, the precise characteristics and functions of EP4 in mollusks remain unclear. In the present study, an EP4 homologue (designated as CgEP4) was identified from oyster Crassostrea gigas. CgEP4 contained a seven-helix transmembrane domain, shared signiﬁcant homology with its vertebrate homologs, and clustered with EP4s from other Mollusca by phylogenetic analysis. When CgEP4 was transfected and expressed in HEK293 cell line, the concentration of intracellular Ca²⁺ was significantly increased after treatment with its agonist PGE2, while that of cAMP was not obviously changed. The mRNA transcripts of CgEP4 were dominantly expressed in haemocytes, which was 28.37-fold (p < 0.05) higher than that in hepatopancreas. By immunofluorescence analysis, CgEP4 was found to be mainly expressed in the agranulocyte subpopulation, and CgEP4⁺ agranulocyte accounted for 18 % of total haemocytes. After Vibrio splendidus stimulation, the mRNA expression of CgEP4 in haemocytes was not obviously changed at the first 12 h, then significantly up-regulated at 24 h, which was 2.7-fold (p < 0.01) of that in control group. After PGE2 treatment in vivo, the phosphorylation of JNK (p-JNK) in haemocytes, but not the phosphorylated ERK (p-ERK), was significantly decreased, which was 0.64-fold (p < 0.05) of that in control group. Simultaneously, the mRNA expression levels of CgTNF-1 and CgTNF-2 in haemocytes dramatically down-regulated, which were 0.06-fold (p < 0.05) and 0.73-fold (p < 0.05) of that in control group at 24 h. When the EP4 antagonist Grapiprant was added with PGE2 treatment in vivo, the p-JNK in haemocytes significantly increased, concomitant with the up-regulation of expressions of CgTNF-1 and CgTNF-2, which were 2.86-fold (p < 0.05) and 1.31-fold (p < 0.05) of that in control group at 24 h. Collectively, these results provides the experimental evidence of a haemocyte highly-expressed EP4 receptor CgEP4 regulating TNFs’ expression through MAPK pathway in the innate immune response in C. gigas, and it could be used as a surface marker to type and sort a subset of agranulocyte subpopulation among haemocytes.

前列腺素E2通过作用于四种不同的e型前列腺素受体亚型（EP1-EP4），对多种细胞具有不同的生理作用，其中EP4是已知介导哺乳动物单核细胞和巨噬细胞免疫应答的亚型之一。然而，EP4在软体动物中的确切特征和功能尚不清楚。本研究从长牡蛎中鉴定出一种EP4同源物（命名为CgEP4）。CgEP4具有7螺旋跨膜结构域，与脊椎动物同源物具有显著的同源性，并与其他软体动物的ep4聚类。CgEP4转染HEK293细胞系表达后，其激动剂PGE2处理后细胞内Ca2+浓度显著升高，cAMP浓度变化不明显。CgEP4 mRNA转录本主要在血细胞中表达，是肝胰腺的28.37倍（p < 0.05）。免疫荧光分析发现CgEP4主要表达于粒细胞亚群，CgEP4+粒细胞占总血细胞的18%。脾弧菌刺激后，血细胞中CgEP4 mRNA表达在前12 h无明显变化，在24 h显著上调，是对照组的2.7倍（p < 0.01）。体内PGE2处理后，血细胞中JNK （p-JNK）磷酸化水平显著降低，ERK （p-ERK）磷酸化水平不显著降低，是对照组的0.64倍（p < 0.05）。同时，血细胞中CgTNF-1和CgTNF-2 mRNA表达量显著下调，分别为0.06倍（p < 0.05）和0.73倍（p < 0.05）

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引用次数: 0