Developmental and comparative immunology最新文献_第8页

Interactions between microbiota and immunity shape pathogen acquisition and fitness in Amblyomma spp. ticks 蜱弱肉病菌群与免疫的相互作用影响病原体的获取和适应。

IF 2.4 3区农林科学 Q1 FISHERIES

Developmental and comparative immunology

Pub Date : 2025-10-13 DOI: 10.1016/j.dci.2025.105493

Solange C. Antão , Daniel B. Pavanelo , Eliane Esteves , Marcelly B. Nassar , Beatriz I. Alonso , Pablo Vera , Marcelo B. Labruna , Petr Kopáček , Sirlei Daffre , Ludek Zurek , Fernanda Dias da Silva , Marisa Farber , Andréa C. Fogaça

Besides carrying pathogens, ticks also harbor commensal and mutualistic microorganisms that constitute their microbiota. This microbial community can modulate the tick immune system and influence pathogen acquisition, either facilitating or hindering colonization. Additionally, the microbiota may impact tick fitness. Although the ticks Amblyomma sculptum and Amblyomma aureolatum are important vectors of Rickettsia rickettsii, the causative agent of Brazilian spotted fever, A. sculptum is much less susceptible to infection than A. aureolatum. Intriguingly, while A. aureolatum midgut harbors an abundant microbiota, mostly composed of bacteria of the Francisella genus, A. sculptum presents a markedly reduced bacterial community. In the current study, we quantified the total bacterial load also in the salivary glands and ovaries of adult A. sculptum and A. aureolatum, besides midgut. Across all analyzed organs, bacterial loads were consistently lower in A. sculptum than in A. aureolatum, regardless of whether the ticks had fed on naïve or R. rickettsii-inoculated hosts. High-throughput sequencing of the V3-V4 hypervariable region of the bacterial 16S rRNA gene revealed that Francisella endosymbiont is the dominant taxon in all organs of control A. aureolatum, with the highest relative frequency in the ovaries and the lowest in the midgut. The highest relative frequency of Francisella in the ovaries correlates with the lower susceptibility of this organ to R. rickettsii, suggesting that the endosymbiosis may limit infection. No 16S rRNA gene sequences could be obtained for A. sculptum samples, likely due to their low bacterial content. To investigate the role played by the microbiota on rickettsial acquisition and tick fitness, A. aureolatum engorged females were treated with either tetracycline or ciprofloxacin. Tetracycline treatment significantly reduced bacterial loads and antimicrobial peptide transcript levels in the eggs, and this was followed by a higher acquisition of R. rickettsii by hatched larvae. Additionally, tetracycline negatively impacted tick development, reducing the molt success from the larval to the nymphal stage. These results suggest that maternal microbiota plays a role in shaping offspring immunity, pathogen susceptibility, and tick development. The multifaceted role of tick microbiota in both development and vector competence underscores its potential as a biotechnological resource for developing new strategies to control tick-borne diseases.

除了携带病原体外，蜱还携带共生和互惠的微生物，这些微生物构成了它们的微生物群。这种微生物群落可以调节蜱的免疫系统，影响病原体的获取，促进或阻碍定植。此外，微生物群可能会影响蜱虫的适应性。虽然壁虱雕刻无形体和金黄色无形体是巴西斑疹热病原体立克次体的重要媒介，但雕刻无形体比金黄色无形体不易感染。有趣的是，虽然金色假单胞菌中肠含有丰富的微生物群，主要由Francisella属细菌组成，但雕塑假单胞菌的细菌群落明显减少。在目前的研究中，除了中肠外，我们还量化了成年雕塑假单胞菌和金黄色假单胞菌的唾液腺和卵巢中的细菌总负荷。在所有被分析的器官中，无论蜱虫是否以naïve或接种了立克次体的宿主为食，雕塑蜱的细菌负荷始终低于金黄色蜱。细菌16S rRNA基因V3-V4高变区高通量测序结果显示，Francisella内共生菌是金色小蠊各器官的优势分类群，其在卵巢的相对频率最高，在中肠的相对频率最低。卵巢中弗兰西斯菌的最高相对频率与该器官对立克次体的较低易感性相关，表明内共生可能限制感染。A. sculptum样品未获得16S rRNA基因序列，可能是由于其细菌含量低。为了研究微生物群对立克次体获得和蜱适合度的影响，研究人员分别用四环素和环丙沙星处理金黄色小蠊。四环素处理显著降低了鸡蛋中的细菌负荷和抗菌肽转录水平，随后孵化的幼虫获得了更高的立克次体。此外，四环素对蜱的发育有负面影响，降低了从幼虫到若虫阶段的蜕皮成功率。这些结果表明，母体微生物群在塑造后代免疫力、病原体易感性和蜱发育中起着重要作用。蜱微生物群在发展和媒介能力方面的多方面作用强调了其作为制定控制蜱传疾病新战略的生物技术资源的潜力。

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引用次数: 0

LvHcS52, a Litopenaeus vannamei hemocyanin-derived peptide, restricts WSSV infection by promoting phagocytosis and activating the STAT signaling pathway LvHcS52是凡纳滨对虾血青素衍生肽，通过促进吞噬和激活STAT信号通路来限制WSSV感染。

IF 2.4 3区农林科学 Q1 FISHERIES

Developmental and comparative immunology

Pub Date : 2025-10-11 DOI: 10.1016/j.dci.2025.105490

Shixiong Zhan , Defu Yao , Yueling Zhang

Shrimp hemocyanin plays multifunctional roles in immunity, both in its native form and as proteolytic fragments generated during infection. Here, we characterize LvHcS52, a hemocyanin-derived peptide identified in the serum of white spot syndrome virus (WSSV)-infected Litopenaeus vannamei. Through molecular weight analysis, N-terminal and de novo sequencing, and bioinformatic prediction, we define LvHcS52 as a 55-amino acid fragment (residues 526–580 of hemocyanin; accession CAA57880) with a molecular mass of 5824.09 Da, structurally featuring one α-helix and two β-strands. Both recombinant and synthetic LvHcS52 significantly inhibited WSSV gene expression (ie1 and vp28), reduced viral loads, and improved shrimp survival. Mechanistically, LvHcS52 binds the WSSV envelope protein VP28 and host β-integrin on hemocytes, promoting viral phagocytosis. Moreover, it also activates the STAT signaling pathway and upregulates anti-lipopolysaccharide factors (ALFs) to restrict viral replication. Our findings underscore the immunological versatility of hemocyanin and illustrate how invertebrates maximize limited immune components to mount effective antiviral responses. This study provides new insights into crustacean innate immunity and opens avenues for developing peptide-based antiviral strategies in aquaculture.

虾血青素在免疫中发挥着多种功能，既以其天然形式存在，也作为感染过程中产生的蛋白水解片段存在。在这里，我们鉴定了LvHcS52，一种在白斑综合征病毒（WSSV）感染的凡纳滨对虾（Litopenaeus vannamei）血清中发现的血青素衍生肽。通过分子量分析、n端和从头测序以及生物信息学预测，我们将LvHcS52定义为一个55个氨基酸的片段（血青素残基526-580，编号CAA57880），分子量为5824.09 Da，结构上具有1个α-螺旋和2个β-链。重组和合成LvHcS52均能显著抑制WSSV基因（ie1和vp28）的表达，降低病毒载量，提高对虾存活率。在机制上，LvHcS52结合WSSV包膜蛋白VP28和宿主β-整合素在血细胞上，促进病毒吞噬。此外，它还激活STAT信号通路，上调抗脂多糖因子（anti- lipopolaccharfactors, ALFs），限制病毒复制。我们的研究结果强调了血青素的免疫多功能性，并说明了无脊椎动物如何最大限度地利用有限的免疫成分来产生有效的抗病毒反应。该研究为甲壳类动物的先天免疫提供了新的见解，并为水产养殖中基于肽的抗病毒策略的开发开辟了道路。

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引用次数: 0

Persistence of viral load in shrimp that survived WSSV infection 存活WSSV感染的虾体内病毒载量的持久性。

IF 2.4 3区农林科学 Q1 FISHERIES

Developmental and comparative immunology

Pub Date : 2025-10-10 DOI: 10.1016/j.dci.2025.105488

Phasini Buathongkam , Jiraporn Srisala , Chanisara Srivihok , Sithichoke Tangphatsornruang , Christopher J. Coates , Suparat Taengchaiyaphum , Siripong Thitamadee , Kallaya Sritunyalucksana

We established a cohabitation model to study shrimp survival after a white spot syndrome virus (WSSV) outbreak. Naïve shrimp were reared individually in plastic boxes immersed in a tank with ten free-roaming shrimp injected each with 1000 copies of purified WSSV. A WSSV outbreak commenced from day four (elevated mortality levels), which lasted for about 10 days. When no further mortalities occurred, surviving shrimp were collected for observation. Survival levels of the cohabitating shrimp were between 5.3 % and 15.9 % from independent infection trials. Determination of viral loads by qPCR and RT-PCR demonstrated 10,000-fold higher viral copy numbers in the moribund shrimp than in the survivors. Western blot analysis using an anti-VP28 antibody confirmed PCR results that high VP28 expression occurs in moribund shrimp, but no signals were detected in the survivors. Histological examination depicted eosinophilic inclusion bodies with hypertrophied (swollen) nuclei and marginated, slightly basophilic, chromatin in the moribund shrimp, but not in the survivors. These data suggest that the surviving shrimp are resilient and posses a mechanism to curtail viremia. Expression levels of selected antimicrobial factors – ALF3, ALF6, PmCrustin1, PmPenaeidin3 and PmPenaeidin5 were compared between moribund and survivor shrimp. PmCrustin1 and ALF3 expression were substantially higher in the moribund shrimp than those of survivors. Interestingly, expression levels of PmCrustin1 were correlated positively with viral loads. Our data provides new insight into WSSV resilience in Penaeus monondon.

为了研究白斑综合征病毒（WSSV）爆发后对虾的生存状况，建立了一种同居模型。Naïve虾被单独饲养在塑料箱中，浸泡在一个水箱中，10只自由游动的虾每只注射1000份纯化的WSSV。WSSV疫情从第4天开始（死亡率升高），持续了大约10天。当没有进一步死亡时，收集幸存虾进行观察。在独立感染试验中，同居对虾的存活率在5.3% ~ 15.9%之间。通过qPCR和RT-PCR对病毒载量的测定表明，死虾的病毒拷贝数比存活虾高10000倍。使用抗VP28抗体的Western blot分析证实了PCR结果，VP28在死虾中有高表达，而在存活虾中没有检测到信号。组织学检查显示，濒死虾中有嗜酸性包涵体，细胞核肥大（肿胀），染色质边缘有轻微的嗜碱性，但幸存者中没有。这些数据表明，幸存的虾具有弹性，并具有减少病毒血症的机制。比较了死虾和活虾中筛选出的抗菌因子ALF3、ALF6、PmCrustin1、PmPenaeidin3和PmPenaeidin5的表达水平。PmCrustin1和ALF3在濒死虾中的表达明显高于存活虾。有趣的是，PmCrustin1的表达水平与病毒载量呈正相关。我们的数据为白对虾（Penaeus monondon）的WSSV恢复能力提供了新的见解。

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引用次数: 0

Molecular characterization and antioxidant function of glutaredoxin 3 in Sepiella japonica 稻瘟病菌谷氨酰胺还毒素3的分子特征及抗氧化功能。

IF 2.4 3区农林科学 Q1 FISHERIES

Developmental and comparative immunology

Pub Date : 2025-10-08 DOI: 10.1016/j.dci.2025.105487

Yan Lu , Peipei Fu , Jiaxin Gao , Yingying Ye , Xinqin Jiang , Denghui Zhu

Glutaredoxins (Grxs) are essential antioxidant proteins that protect cells from oxidative damage and participate in multiple biological processes. However, their roles in the common Chinese cuttlefish (Sepiella japonica) remain uncharacterized. Here, we identified a glutaredoxin 3 homolog from S. japonica (SjGrx3). The SjGrx3 gene encodes a 981 bp open reading frame producing a 326-amino-acid protein (36.47 kDa) that contains an N-terminal thioredoxin domain and two C-terminal Grx domains, each containing the conserved CGFS active-site motif. Phylogenetic analysis clustered SjGrx3 within the cephalopod Grx3 clade, showing high sequence identity to other molluscan homologs. The SjGrx3 mRNA was detected in all examined tissues, with the highest levels in skin and kidney and moderate levels in gill, gut, and liver. In naturally Vibrio harveyi-infected individuals, SjGrx3 transcripts increased significantly in skin, gill, liver, white body, and brain, implicating it in innate immune responses. Confocal microscopy showed that SjGrx3 localized to both the cytoplasm and nucleus. Functional assays demonstrated that recombinant SjGrx3 was soluble and enhanced Escherichia coli resistance to H₂O₂-induced oxidative damage. Moreover, overexpression of SjGrx3 in HEK293T cells significantly reduced H₂O₂-induced intracellular reactive oxygen species (ROS) accumulation and inhibited H₂O₂-mediated cell death, confirming its role in oxidative stress protection. Collectively, these findings indicate that SjGrx3 contributes to redox regulation and immune defense against bacterial infection in S. japonica.

Glutaredoxins （Grxs）是一种重要的抗氧化蛋白，可保护细胞免受氧化损伤并参与多种生物过程。然而，它们在普通中国墨鱼（Sepiella japonica）中的作用尚未明确。在此，我们从粳稻中鉴定了一个glutaredoxin 3同源物（SjGrx3）。SjGrx3基因编码一个981 bp的开放阅读框，产生一个326个氨基酸的蛋白（36.47 kDa），包含一个n端硫氧还蛋白结构域和两个c端Grx结构域，每个结构域都包含保守的CGFS活性位点基序。系统发育分析将SjGrx3聚集在头足类Grx3分支中，显示出与其他软体动物同源物的高度序列一致性。SjGrx3 mRNA在所有检查组织中均检测到，皮肤和肾脏中含量最高，鳃、肠道和肝脏中含量中等。在自然感染哈维弧菌的个体中，SjGrx3转录本在皮肤、鳃、肝脏、白体和大脑中显著增加，暗示其参与先天免疫反应。共聚焦显微镜显示SjGrx3定位于细胞质和细胞核。功能分析表明，重组SjGrx3具有可溶性，增强了大肠杆菌对h2o2诱导的氧化损伤的抗性。此外，在HEK293T细胞中过表达SjGrx3可显著降低h2o2诱导的细胞内活性氧（ROS）积累，抑制h2o2介导的细胞死亡，证实其在氧化应激保护中的作用。综上所述，这些发现表明SjGrx3参与了粳稻的氧化还原调控和对细菌感染的免疫防御。

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引用次数: 0

Identification, evolutionary analysis and expression patterns of the Interleukin-10 superfamily in Megalobrama amblycephala 白介素-10超家族的鉴定、进化分析及表达模式。

IF 2.4 3区农林科学 Q1 FISHERIES

Developmental and comparative immunology

Pub Date : 2025-10-08 DOI: 10.1016/j.dci.2025.105468

Zhensheng Wang , Wenya Zhai , Zhenling Duan , Mengfan Li , Zexia Gao , Huanling Wang , Hong Liu

The interleukin-10 superfamily members (IL-10s) play a crucial role in defending against infections by pathogenic microorganisms. Meanwhile, their significance in the immune response of fish against diseases is well recognized, yet systematic analyses of the IL-10s in fish remain limited. In this study, we identified six IL-10s (Mail-10a, Mail-10b, Mail-10c, Mail-19l, Mail-22, and Mail-26) in the Megalobrama amblycephala genome. Phylogenetic analyses indicated that M. amblycephala IL-10s (MaIL-10s) were conserved in vertebrates. Meanwhile, linear analyses indicated that Mail-10a, Mail-22, and Mail-26 exhibited conserved genetic environments in representative species. In addition, syntenic analysis suggested that segmental duplication events were instrumental in the evolutionary expansion of MaIL-10s. Within the MaIL-10s, substantial variations in conserved motifs, gene structure, and protein structure were observed, indicating its complex evolutionary history. Expression profiling revealed high expression of Mail-10 analogues (Mail-10a, Mail-10b, and Mail-10c) in professional immune organs (spleen, trunk kidney, and head kidney), but low expression in the intestine. In contrast, Mail-22 showed the highest expression in mucosal tissues (intestine and gill). Mail-19l and Mail-26 exhibited the highest expression in blood. After Aeromonas hydrophila infection, MaIL-10s displayed tissue-specific and time-specific expression patterns in infected tissues (intestine, spleen, liver). In summary, our findings deepen the understanding of the IL-10 superfamily and lay the groundwork for exploring the functions of MaIL-10s in bacterial diseases.

白细胞介素-10超家族成员（il -10）在抵御病原微生物感染中起着至关重要的作用。同时，il -10在鱼类疾病免疫应答中的重要作用已得到公认，但对鱼类il -10的系统分析仍然有限。在这项研究中，我们鉴定了6个il-10 （Mail-10a、Mail-10b、Mail-10c、mail - 191、Mail-22和Mail-26）。系统发育分析表明，M. amblycephala IL-10s （MaIL-10s）在脊椎动物中具有保守性。同时，线性分析表明Mail-10a、Mail-22和Mail-26在代表性物种中表现出保守的遗传环境。此外，综合分析表明，片段重复事件在mail -10的进化扩展中发挥了重要作用。在MaIL-10s中，保守基序、基因结构和蛋白质结构发生了实质性的变化，表明其复杂的进化历史。表达谱显示Mail-10类似物（Mail-10a、Mail-10b和Mail-10c）在专业免疫器官（脾、干肾和头肾）中高表达，但在肠道中低表达。Mail-22在粘膜组织（肠和鳃）中表达量最高。mail - 191和Mail-26在血液中表达量最高。在嗜水气单胞菌感染后，MaIL-10s在感染组织（肠、脾、肝）中表现出组织特异性和时间特异性的表达模式。总之，我们的发现加深了对IL-10超家族的认识，为探索mail -10在细菌性疾病中的功能奠定了基础。

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引用次数: 0

Effects of feed supplementation with Lactobacillus reuteri and Bacillus subtilis on the liver immunity in largemouth bass (Micropterus salmoides) 饲料中添加罗伊氏乳杆菌和枯草芽孢杆菌对大口黑鲈肝脏免疫的影响。

IF 2.4 3区农林科学 Q1 FISHERIES

Developmental and comparative immunology

Pub Date : 2025-10-06 DOI: 10.1016/j.dci.2025.105486

Chong Wang , Xiaodi Hu , Huijuan Tang , Wei Ge , Lijun Di , Jixing Zou , Aiguo Zhou , Zongbin Cui

In the culture of largemouth bass (Micropterus salmoides), probiotics play an important role. Probiotics such as Lactobacillus reuteri and Bacillus subtilis can enhance the immune systems of fish like M. salmoides and help them prevent diseases. The largemouth bass in this experiment were fed a diet containing B. subtilis (BS, 10⁷ CFU/g) and L. reuteri (LR, 10⁷ CFU/g) for 35 days. Investigations were conducted into how the two probiotics affected the liver antioxidant capacity and liver immunity of M. salmoides. The findings revealed that both the BS and LR groups activated the Nrf2/keap1 signaling pathway, significantly increasing the expression of genes such as Nrf2, SOD1, SOD2, GST, and CAT. Moreover, the BS group notably augmented the levels of AKP, T-AOC, SOD, and CAT in the liver. Importantly, the key immune-related pathways enriched for differentially expressed genes in the BS group for hepatic transcriptional machinery encompassed complement and coagulation cascades, IL-17 signaling pathway, and chemokine signaling pathway. In conclusion, B. subtilis and L. reuteri demonstrate the ability to enhance the hepatic antioxidant capacity and immunocompetence of M. salmoides. However, significant differences exist in the intensity of their effects and the types of immune pathways activated in the liver. Both hold promising application value, and this study lays a foundational groundwork for the utilization of B. subtilis and L. reuteri in M. salmoides aquaculture.

在大口黑鲈（Micropterus salmoides）养殖中，益生菌起着重要的作用。益生菌如罗伊氏乳杆菌和枯草芽孢杆菌可以增强鱼的免疫系统，帮助它们预防疾病。本试验用含枯草芽孢杆菌（BS, 107 CFU/g）和罗伊氏乳杆菌（LR, 107 CFU/g）的饲料饲喂大口黑鲈35 d。研究了两种益生菌对沙门菌肝脏抗氧化能力和肝脏免疫功能的影响。结果显示，BS和LR组均激活了Nrf2/keap1信号通路，显著增加了Nrf2、SOD1、SOD2、GST和CAT等基因的表达。此外，BS组显著提高了肝脏中AKP、T-AOC、SOD和CAT的水平。重要的是，BS组中差异表达基因富集的关键免疫相关通路包括补体和凝血级联、IL-17信号通路和趋化因子信号通路。综上所述，枯草芽孢杆菌和罗伊氏乳杆菌均能增强沙门沙门的肝脏抗氧化能力和免疫能力。然而，它们的作用强度和肝脏中激活的免疫途径类型存在显着差异。两者均具有良好的应用价值，本研究为枯草芽孢杆菌和罗伊氏乳杆菌在鲑鱼养殖中的利用奠定了基础。

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引用次数: 0

Lamprey TFE3 exhibits evolutionarily conserved activation mechanisms and regulates autophagy and immune responses 七鳃鳗TFE3表现出进化保守的激活机制，并调节自噬和免疫反应。

IF 2.4 3区农林科学 Q1 FISHERIES

Developmental and comparative immunology

Pub Date : 2025-10-04 DOI: 10.1016/j.dci.2025.105480

Jilong Pan , Xinping Li , Qipeng Zhang , Jiarui Li , Daiyun Zhang , Xiaoyu Ni , Pengcheng Li , Tiesong Li , Hao Wang , Yan Chi

The microphthalmia-associated transcription factor/transcription factor E (MiT/TFE) family belongs to the basic group of helix-loop-helix leucine zipper (bHLH-ZIP)-containing transcription factors. Although MiT/TFE family members, especially TFEB and TFE3, have been extensively studied in higher vertebrates, little is known about their roles in basal vertebrates such as lampreys. Here, we show that lamprey TFE3 and TFEC retain conserved structural features, including key domains, motifs, and serine residues, consistent with the evolutionary history of vertebrates. Expression profiling revealed that lamprey TFE3 is broadly expressed across tissues, with the highest level in the heart. Treatment with Torin1 induced nuclear translocation of TFE3, indicative of its activation. Torin1 also led to a time-dependent increase in TFE3 expression, and upregulation of downstream target genes associated with autophagy, stress responses, and inflammation. Furthermore, we observed elevated LC3-II levels and reduced p62 expression following Torin1 treatment, indicating that activated lamprey TFE3 enhances autophagic activity. Notably, TFE3 activation also promoted cholesterol mobilization and efflux, as evidenced by decreased Bodipy fluorescence intensity and upregulated expression of ABCA1. In vivo stimulation with LPS or poly (I:C) induced significant changes in Lj-TFE3 expression, indicating that lamprey TFE3 is responsive to pathogen-associated molecular patterns and may participate in immune stress responses. Together, these findings demonstrate that lamprey TFE3 exhibits conserved structural and functional features and plays a key role in immune and metabolic regulation, providing important evolutionary insights into the MiT/TFE transcription factor family in early vertebrates.

小眼相关转录因子/转录因子E （MiT/TFE）家族属于含有螺旋-环-螺旋亮氨酸拉链（bHLH-ZIP）的转录因子的基本类群。虽然MiT/TFE家族成员，特别是TFEB和TFE3，在高等脊椎动物中被广泛研究，但对它们在基底脊椎动物（如七鳃鳗）中的作用知之甚少。本研究表明，七鳃鳗TFE3和TFEC保留了保守的结构特征，包括关键结构域、基序和丝氨酸残基，与脊椎动物的进化史一致。表达谱分析显示，七鳃鳗TFE3在各组织中广泛表达，在心脏中表达水平最高。Torin1诱导TFE3核易位，表明其活化。Torin1还导致TFE3表达的时间依赖性增加，以及与自噬、应激反应和炎症相关的下游靶基因的上调。此外，我们观察到在Torin1处理后，LC3-II水平升高，p62表达降低，表明激活的TFE3增强了七鳃鳗的自噬活性。值得注意的是，TFE3的激活也促进了胆固醇的动员和外排，Bodipy荧光强度降低，ABCA1表达上调。体内LPS或poly （I:C）刺激诱导Lj-TFE3表达显著变化，提示七鳃鳗TFE3响应病原体相关分子模式，可能参与免疫应激反应。总之，这些发现表明，七鳃鳗TFE3具有保守的结构和功能特征，并在免疫和代谢调节中发挥关键作用，为早期脊椎动物的MiT/TFE转录因子家族提供了重要的进化见解。

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引用次数: 0

Preparation and characterization of monoclonal antibodies against porcine caspase-7 猪caspase-7单克隆抗体的制备与鉴定。

IF 2.4 3区农林科学 Q1 FISHERIES

Developmental and comparative immunology

Pub Date : 2025-10-04 DOI: 10.1016/j.dci.2025.105485

Chenyi Guo , Lanjie Yu , Shuangshuang Zhang , Yongsheng Cao , Wenlong Zhang

Cysteine-aspartic proteases (Caspases) play a central role in programmed cell death (PCD). Caspase-7 is an effector caspase involved in the execution phase of apoptosis and pyroptosis. Determining the expression and activation of caspase-7 is often an essential experiment in studies on PCD. Currently, commercially available antibodies specific to human and mouse caspase-7 have facilitated research on PCD in these species. A specific antibody against porcine caspase-7 (pcaspase-7) is needed, as PCD has been closely linked to porcine diseases. In this study, a recombinant pcaspase-7 protein (rpcaspase-7) with cysteine replaced by alanine at the 186 position was prokaryotically expressed and purified. This recombinant protein was used to immunize BALB/c mice to generate pcaspase-7-specific monoclonal antibodies (mAbs). The prepared 7-4E1-F4-D2-E1 mAb targets the 90–100 amino acid (aa) oligopeptide (TDKDAEALFKC) of pcaspase-7, while the mAbs 7-3F1-E2-H2-D6, 7-3F1-E2-H2-D1, 7-4E5-C11-C9-B5, and 7-4E5-C11-C9-F11 target the 45–55 aa oligopeptide (GSSVKIPRDRE). All prepared mAbs can discriminate pcaspase-7 from pcaspase-1, -3, -8, and -13. The mAbs targeting the 45–55 aa oligopeptide can also distinguish pcaspase-7 from human, hamster, and bovine caspase-7. The 7-4E1-F4-D2-E1 mAb recognizes pcaspase-7 in Western blot assays but not in immunofluorescence assays. These mAbs can be used to purify prokaryotically expressed pcaspase-7 via immunoprecipitation and to detect pcaspase-7 activation in virus-infected porcine cells. Thus, they represent valuable tools for future structural and functional studies of pcaspase-7. Additionally, this study demonstrated that pcaspase-7 can cleave porcine GSDMD, revealing a previously unrecognized function of pcaspase-7.

半胱氨酸-天冬氨酸蛋白酶（Caspases）在程序性细胞死亡（PCD）中起核心作用。caspase -7是一种参与细胞凋亡和焦亡执行阶段的效应caspase。确定caspase-7的表达和激活通常是PCD研究中必不可少的实验。目前，市售的针对人和小鼠caspase-7的抗体促进了这些物种PCD的研究。由于PCD与猪疾病密切相关，因此需要一种针对猪caspase-7 （pcaspase-7）的特异性抗体。本研究对重组pcaspase-7蛋白（rpcaspase-7）进行了原核表达和纯化，该蛋白186位的半胱氨酸被丙氨酸取代。利用该重组蛋白免疫BALB/c小鼠产生pcaspase-7特异性单克隆抗体（mab）。制备的7-4E1-F4-D2-E1单抗靶向pcaspase-7的90-100个氨基酸寡肽（TDKDAEALFKC），而7-3F1-E2-H2-D6、7-3F1-E2-H2-D1、7-4E5-C11-C9-B5和7-4E5-C11-C9-F11单抗靶向45-55个氨基酸寡肽（GSSVKIPRDRE）。所有制备的单克隆抗体均能区分pcaspase-7与pcaspase-1、-3、-8和-13。靶向45- 55aa寡肽的单抗也能区分人、仓鼠和牛的caspase-7。7-4E1-F4-D2-E1 mAb在western blot检测中识别pcaspase-7，但在免疫荧光检测中不识别。这些单抗可以通过免疫沉淀纯化原核表达的pcaspase-7，并检测病毒感染的猪细胞中pcaspase-7的激活情况。因此，它们为未来pcaspase-7的结构和功能研究提供了有价值的工具。此外，本研究表明pcaspase-7可以切割猪GSDMD，揭示了pcaspase-7以前未被认识的功能。

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引用次数: 0

A review of host and pathogen gene targets for RNAi therapeutics in shrimp disease mitigation RNAi治疗对虾疾病的宿主和病原体基因靶点研究进展

IF 2.4 3区农林科学 Q1 FISHERIES

Developmental and comparative immunology

Pub Date : 2025-09-30 DOI: 10.1016/j.dci.2025.105482

Maria Violeta R. Tare , Debrah Jannsen DJ N. Almazan , Krisha Marie D.L. Saquilayan , Patricia Jhoanna V. Glori , Jim Troy Solitario , Mary Beth B. Maningas

RNA interference (RNAi) has emerged as a powerful therapeutic and research tool in shrimp aquaculture, with its significant potential for disease mitigation. This review explores the strategic targeting of essential genes–either from the host gene or the pathogen–as a basis for RNAi-based therapies in shrimp. Targeting host genes, especially those involved in immune responses, can lead to increased protective effects but also poses risks of lethality if essential physiological pathways are disrupted. Conversely, silencing pathogen genes offers pathogen-specific intervention, though with limitations in broad-spectrum applicability and potential off-target effects. This paper further discusses multi-variant RNAi approaches that target both host and pathogen genes, highlighting their potential in achieving greater protective effects. Also included is a review of Bioinformatics and AI applications in RNAi, and the possibilities it may offer for gene silencing in shrimp. Current evidence suggests that targeting host immune-related genes may yield higher survival outcomes, but a tailored, case-by-case approach remains important. Ultimately, the choice of target depends on a nuanced understanding of host-pathogen interactions. The findings underscore the need for continued research to optimize RNAi strategies for sustainable shrimp disease management.

RNA干扰（RNAi）已成为虾类养殖中一种强大的治疗和研究工具，具有显著的疾病缓解潜力。本文综述了从宿主基因或病原体中靶向必需基因的策略，以作为虾类rnai治疗的基础。以宿主基因为目标，特别是那些参与免疫反应的基因，可导致保护作用增强，但如果基本的生理途径被破坏，也会带来致命的风险。相反，沉默病原体基因提供了病原体特异性干预，尽管在广谱适用性和潜在的脱靶效应方面存在局限性。本文进一步讨论了针对宿主和病原体基因的多变体RNAi方法，强调了它们在实现更大保护作用方面的潜力。还包括生物信息学和人工智能在RNAi中的应用，以及它可能为虾的基因沉默提供的可能性。目前的证据表明，靶向宿主免疫相关基因可能会产生更高的生存结果，但一个量身定制的，具体情况具体分析的方法仍然很重要。最终，靶标的选择取决于对宿主-病原体相互作用的细微理解。这些发现强调了继续研究优化RNAi策略以实现对虾疾病可持续管理的必要性。

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引用次数: 0

Non-lethal heat shock enhances the immune response of Procambarus clarkii hemocytes to Vibrio parahaemolyticus through the HSP90 gene family 非致死性热休克通过HSP90基因家族增强了克氏原螯虾血细胞对副溶血性弧菌的免疫应答。

IF 2.4 3区农林科学 Q1 FISHERIES

Developmental and comparative immunology

Pub Date : 2025-09-30 DOI: 10.1016/j.dci.2025.105484

Shirui Yue , Xiuhong Cai , Zhangxuan Chen , Yuyan Gong , Yuqing Liu , Bao Wang , Yewen Xi , Shunchang Wang , Xin Zhang

This research focuses on Procambarus clarkii and systematically explores the mechanism of action of the HSP90 gene family when responding to non-lethal heat shock (NLHS) and Vibrio parahaemolyticus infection. We have identified four members of the HSP90 gene family. The characteristics of the proteins encoded by these genes are significantly different, which implies functional divergence. Multiple sequence alignment and phylogenetic analysis suggested that the structure of HSP90 proteins exhibits both conservatism and diversity, indicating functional divergence among family members. The genes are unevenly distributed across chromosomes and there are no tandem repeat genes. HSP90 genes are differentially expressed in tissues, with high expression in hemocytes indicating immune involvement. After NLHS, the expression of some genes increased dramatically and reached the peak after injection with V. parahaemolyticus. However, for some other genes, their expression did not change significantly, indicating that they may be involved in different types of regulation. Protein interaction and co-expression analyses revealed HSP90 proteins interact with innate immune proteins and collaborate with TLR pathway members. DsRNA silencing of HSP90 downregulates key TLR molecules (TLR2, TRAF6, IRAK4, MyD88) at different time points, confirming their functional immune association. This indicates HSP90 genes play a key role in anti-pathogen mechanisms. The study provides molecular markers for breeding stress-resistant P. clarkii and theoretical support for crustacean stress biology research.

本研究以克氏原螯虾为研究对象，系统探讨了HSP90基因家族对非致死性热休克（NLHS）和副溶血性弧菌感染的作用机制。我们已经确定了HSP90基因家族的四个成员。这些基因编码的蛋白质特征有显著差异，这意味着功能上的差异。多序列比对和系统发育分析表明，HSP90蛋白结构具有保守性和多样性，表明家族成员之间存在功能分化。基因在染色体上分布不均匀，不存在串联重复基因。HSP90基因在组织中有差异表达，在血细胞中的高表达表明免疫参与。NLHS后，部分基因表达量显著增加，注射副溶血性弧菌后表达量达到峰值。然而，对于其他一些基因，它们的表达没有明显变化，表明它们可能参与不同类型的调控。蛋白相互作用和共表达分析显示，HSP90蛋白与先天免疫蛋白相互作用，并与TLR通路成员协同作用。DsRNA沉默HSP90在不同时间点下调关键TLR分子（TLR2、TRAF6、IRAK4、MyD88），证实了它们的功能性免疫关联。这表明HSP90基因在抗病原体机制中起关键作用。该研究为克氏假单胞菌的抗性育种提供了分子标记，并为甲壳类动物的逆境生物学研究提供了理论支持。

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引用次数: 0