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Characterization of NLRP3 inflammasome components in the endangered Chinese giant salamander (Andrias davidianus) 濒危中国大鲵体内 NLRP3 炎症小体成分的特征描述
IF 2.7 3区 农林科学 Q1 FISHERIES Pub Date : 2024-09-10 DOI: 10.1016/j.dci.2024.105263
Xuan Wei , Jianxiong Wu , Xiangyu Pi , Qihuan Zhang , Jingyu Tian , Zhitao Qi

Chinese giant salamander (Andrias davidianus) is the largest extant urodela species and has unique evolutionary position. Studying the immune system of Chinese giant salamander contributes to understanding the evolution of immune systems of vertebrates. The NLR-related protein 3 (NLRP3) inflammasome comprised of NLRP3, ASC and caspase-1 play important roles in the host innate immunity. However, little is know about the NLRP3 inflammasome components in Chinese giant salamander. In this study, the NLRP3, apoptosis-associated speck-like protein (ASC) and caspase-1 (adaNLRP3, adaASC and adaCaspase-1) were characterized from Chinese giant salamander. The proteins of these three genes shared similar motifs and structures with their mammalian counterparts, with a PYD motif, a nucleotide-binding domain (NACHT) motif, and four leucine-rich repeat domain (LRR) motifs identified in adaNLRP3, a pyrin domain (PYD) motif and a caspase recruitment domain (CARD) motif in adaASC, and a CARD motif and a CASc motif in adaCaspase-1. These three genes were constitutively expressed in the skin, heart, lung, kidney, muscle, brain, spleen, and liver of Chinese giant salamander. Following Aeromonas hydrophia infection, all the three genes were up-regulated in various tissues. Molecular docking analysis revealed that the key residues involved in forming the adaNLRP3/adaASC complex were located in the PYD motifs, and that involved in forming the adaASC/adaCaspase-1 complex were located in the CARD motifs. Further analysis revealed that the hydrogen bonds and salt bridges had crucial roles in the formation of adaNLRP3/acaASC and adaASC/adaCaspase-1 complexes. To the best of our knowledge, this is the first report on the NLRP3 inflammasome components in Chinese giant salamander which will be helpful in further understanding the function of the NLRP3 inflammasome and in elucidating its role in the immune response to microbes.

中国大鲵(Andrias davidianus)是现存最大的蝾螈类物种,具有独特的进化地位。研究中国大鲵的免疫系统有助于了解脊椎动物免疫系统的进化。由NLRP3、ASC和caspase-1组成的NLR相关蛋白3(NLRP3)炎性体在宿主先天性免疫中发挥着重要作用。然而,人们对中国大鲵体内的NLRP3炎性体成分知之甚少。本研究对中国大鲵的NLRP3、凋亡相关斑点样蛋白(ASC)和caspase-1(adaNLRP3、adaASC和adaCaspase-1)进行了鉴定。这三个基因的蛋白与哺乳动物的蛋白具有相似的基序和结构,在adaNLRP3中发现了一个PYD基序、一个核苷酸结合结构域(NACHT)基序和四个富亮氨酸重复结构域(LRR)基序;在adaASC中发现了一个吡啶结构域(PYD)基序和一个caspase招募结构域(CARD)基序;在adaCaspase-1中发现了一个CARD基序和一个CASc基序。这三个基因在中国大鲵的皮肤、心脏、肺、肾脏、肌肉、脑、脾脏和肝脏中均呈组成型表达。感染嗜水气单胞菌后,这三个基因在不同组织中均上调。分子对接分析表明,参与形成adaNLRP3/adaASC复合物的关键残基位于PYD基团,而参与形成adaASC/adaCaspase-1复合物的关键残基位于CARD基团。进一步分析发现,氢键和盐桥在adaNLRP3/acaASC和adaASC/adaCaspase-1复合物的形成过程中起着关键作用。据我们所知,这是首次报道中国大鲵的NLRP3炎性体成分,这将有助于进一步了解NLRP3炎性体的功能,并阐明其在微生物免疫反应中的作用。
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引用次数: 0
First report of a chemokine from camelids: Dromedary CXCL8 is induced by poxvirus and heavy metal toxicity 首次报道驼科动物的趋化因子:单峰驼的 CXCL8 由痘病毒和重金属毒性诱导。
IF 2.7 3区 农林科学 Q1 FISHERIES Pub Date : 2024-09-05 DOI: 10.1016/j.dci.2024.105261
Avinash Premraj, Abi George Aleyas, Binita Nautiyal, Thaha Jamal Rasool

Low molecular weight proteins, known as chemokines, facilitate the migration and localization of immune cells to the site of infection and injury. One of the first chemokines identified, CXCL8 functions as a key neutrophil activator, recruiting neutrophils to sites of inflammation. Several viral infections, including zoonotic coronaviruses and poxviruses, have been reported to induce the expression of CXCL8. Dromedary camels are known to harbor several potentially zoonotic pathogens, but critical immune molecules such as chemokines remain unidentified. We report here the identification of CXCL8 from the dromedary camel - the first chemokine identified from camelids. The complete dromedary CXCL8 cDNA sequence as well as the corresponding gene sequence from dromedary and two New World camelids - alpaca and llama were cloned. CXCL8 mRNA expression was relatively higher in PBMC, spleen, lung, intestine, and liver. Poly(I:C) and lipopolysaccharide stimulated CXCL8 expression in vitro, while interferon treatment inhibited it. In vitro infection with potentially zoonotic camelpox virus induced the expression of CXCL8 in camel kidney cells. Toxicological studies on camelids have been limited, and no biomarkers have been identified. Hence, we also evaluated CXCL8 mRNA expression as a potential biomarker to assess heavy metal toxicity in camel kidney cells in vitro. CXCL8 expression was increased after in vitro exposure to heavy metal compounds of cobalt and cadmium, suggesting potential utility as a biomarker for renal toxicity in camels. The results of our study demonstrate that camel CXCL8 plays a significant role in immunomodulatory and induced toxicity responses in dromedary camels.

被称为趋化因子的低分子量蛋白质可促进免疫细胞向感染和损伤部位迁移和定位。作为最早发现的趋化因子之一,CXCL8 是一种关键的中性粒细胞激活剂,可将中性粒细胞募集到炎症部位。据报道,包括人畜共患冠状病毒和痘病毒在内的几种病毒感染都会诱导 CXCL8 的表达。众所周知,单峰骆驼携带多种潜在的人畜共患病原体,但关键的免疫分子(如趋化因子)仍未确定。我们在此报告从单峰骆驼中鉴定出了 CXCL8,这是第一个从驼科动物中鉴定出的趋化因子。我们克隆了完整的单峰驼 CXCL8 cDNA 序列以及单峰驼和两种新世界驼科动物(羊驼和美洲驼)的相应基因序列。CXCL8 mRNA在PBMC、脾脏、肺脏、肠道和肝脏中的表达量相对较高。聚(I:C)和脂多糖刺激体外 CXCL8 的表达,而干扰素处理则抑制其表达。体外感染可能是人畜共患的驼痘病毒会诱导骆驼肾细胞中 CXCL8 的表达。对骆驼进行的毒理学研究十分有限,也没有发现生物标志物。因此,我们还评估了 CXCL8 mRNA 的表达,将其作为一种潜在的生物标志物来评估体外骆驼肾细胞的重金属毒性。体外暴露于钴和镉的重金属化合物后,CXCL8的表达增加,这表明它有可能成为骆驼肾毒性的生物标志物。我们的研究结果表明,骆驼 CXCL8 在单峰骆驼的免疫调节和诱导毒性反应中发挥着重要作用。
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引用次数: 0
Enhancement of immunity and disease resistance in Litopenaeus vannamei through injection of tyramine formulated with polyethylene glycol 通过注射用聚乙二醇配制的酪胺增强万年青的免疫力和抗病性。
IF 2.7 3区 农林科学 Q1 FISHERIES Pub Date : 2024-09-03 DOI: 10.1016/j.dci.2024.105260
Cheng-Ying Li , Hsin-Wei Kuo , Winton Cheng

This study investigates the prolonged effect of immune disease resistance in Litopenaeus vannamei through the administration of tyramine (TA) formulated with polyethylene glycol (PEG). Facing the challenges of intensive farming, environmental stress, and global climate changes, innovative approaches to improve shrimp health are essential. The research focuses on the role of biogenic amines in stress response and immune regulation, demonstrating that TA, especially when combined with PEG, significantly prolongs immunity and resistance against Vibrio alginolyticus. The experimental design included administering TA, PEG, and TA-PEG, followed by evaluations of immunity, lactate and glucose levels, and immune-related gene expressions. Results showed notable prolonged effects in total hemocyte count, phenoloxidase activity, and phagocytic activity in the TA-PEG group, indicating enhanced immune activation period. Additionally, the expression of prophenoloxidase system-related genes was significantly upregulated in the TA-PEG group. Furthermore, the TA-PEG group exhibited a significantly higher survival rate in a susceptibility test against V. alginolyticus. The results of this study confirm that the combined use of PEG can effectively extend the immunostimulatory duration of TA.

本研究调查了通过施用与聚乙二醇(PEG)配制的酪胺(TA)对南美白对虾免疫抗病性的延长效果。面对集约化养殖、环境压力和全球气候变化的挑战,采用创新方法改善对虾健康至关重要。研究重点是生物胺在应激反应和免疫调节中的作用,结果表明,TA(尤其是与 PEG 结合使用时)能显著延长免疫力,增强对藻类溶解弧菌的抵抗力。实验设计包括施用 TA、PEG 和 TA-PEG,然后评估免疫力、乳酸和葡萄糖水平以及免疫相关基因的表达。结果显示,TA-PEG 组的血细胞总数、酚氧化酶活性和吞噬细胞活性明显延长,表明免疫激活期延长。此外,TA-PEG 组的丙醇氧化酶系统相关基因表达明显上调。此外,TA-PEG 组在对溶藻病毒的敏感性测试中存活率明显更高。本研究结果证实,联合使用 PEG 可有效延长 TA 的免疫刺激持续时间。
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引用次数: 0
The bright future of developmental and comparative immunology: Views from heroes of the field 发育和比较免疫学的光明前景:该领域英雄们的观点
IF 2.7 3区 农林科学 Q1 FISHERIES Pub Date : 2024-08-30 DOI: 10.1016/j.dci.2024.105257
Martin F. Flajnik
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引用次数: 0
Immunohistochemistry analyses of the abomasal mucosa show differences in cellular-mediated immune responses to Haemonchus contortus infection in resistant and susceptible young lambs 腹腔粘膜的免疫组化分析表明,抗感染羔羊和易感感染羔羊对血包虫病的细胞介导免疫反应存在差异
IF 2.7 3区 农林科学 Q1 FISHERIES Pub Date : 2024-08-30 DOI: 10.1016/j.dci.2024.105259
José Gabriel Gonçalves Lins , Ana Cláudia A. Albuquerque , Helder Louvandini , Alessandro F.T. Amarante

Haemonchus contortus is known for its high pathogenicity in sheep, and the uncontrolled use of anthelmintics resulted in the emergence of multiple drug-resistant populations. Breeding sheep for gastrointestinal nematode resistance is a sustainable alternative to reduce dependence of anthelmintic drugs, and differences in the degree of resistance between breeds have been reported. Here we compare two sheep breeds (Santa Ines and Ile de France), concerning the differences in innate and adaptive immune response involved in the resistance against H. contortus infection. Immunohistochemical analyses of the abomasum were conducted in naïve Santa Ines (n = 14) and Ile de France (n = 12) lambs randomized into four groups: infected Santa Ines (n = 8), non-infected control Santa Ines (n = 6), infected Ile de France (n = 8), and non-infected control Ile de France (n = 4). The infected lambs were initially infected with H. contortus infective larvae at 14 days of age, and multiple infections were conducted every second day until they reached 66 days of age. There was a significant effect (P < 0.001) of the infection with increase in numbers of CD3+ T; CD79α+ B; GATA3+ Th2/ILC2; POU2F3+ tuft cells; FOXP3+ T reg; and IgE + cells in the fundus of the abomasal mucosa in both Santa Ines and Ile de France lambs. Nevertheless, the infected Santa Ines lambs presented the highest averages for CD79α+ B; GATA3+ Th2/ILC; IgE + cells; and POU2F3+ tuft cells and there was a significant association of the breed and infection status with regards to POU2F3+ tuft cells, with the highest mean in the infected Santa Ines group. The infected Santa Ines group had three lambs with high degree of resistance and five lambs that showed a moderate infection. Our results suggest a mechanism of synergistic coordination between different immune-cell types in promoting resistance of suckling lambs under H. contortus infection.

众所周知,绵羊腹腔线虫具有很强的致病性,无节制地使用驱虫药导致了多种抗药性种群的出现。培育具有胃肠道线虫抗药性的绵羊是减少对抗虫药依赖性的一种可持续替代方法,而不同品种之间的抗药性差异已有所报道。在此,我们比较了两个绵羊品种(Santa Ines 和 Ile de France)在先天性免疫反应和适应性免疫反应方面的差异,这两种免疫反应都参与了对线虫感染的抵抗。我们对圣伊内斯(14 只)和法兰西岛(12 只)羔羊的腹腔进行了免疫组化分析,随机分为四组:受感染的圣伊内斯(8 只)、未受感染的圣伊内斯对照组(6 只)、受感染的法兰西岛(8 只)和未受感染的法兰西岛对照组(4 只)。受感染的羔羊在 14 日龄时首次感染传染性伊蚊幼虫,每隔一天进行多次感染,直到羔羊长到 66 日龄。感染对圣伊内斯和法兰西岛羔羊腹腔粘膜底部的 CD3+ T、CD79α+ B、GATA3+ Th2/ILC2、POU2F3+ 簇细胞、FOXP3+ T reg 和 IgE + 细胞数量的增加有明显影响(P < 0.001)。然而,受感染的圣伊内斯羔羊的 CD79α+ B、GATA3+ Th2/ILC、IgE + 细胞和 POU2F3+ 簇细胞的平均值最高。受感染的圣伊内斯组中有三只羔羊表现出高度抵抗力,五只羔羊表现出中度感染。我们的研究结果表明,不同类型的免疫细胞之间存在协同配合机制,可促进哺乳羔羊对传染性软疣的抵抗力。
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引用次数: 0
A personal view on developmental and comparative immunology: What, how and why? 关于发育和比较免疫学的个人观点:内容、方法和原因?
IF 2.7 3区 农林科学 Q1 FISHERIES Pub Date : 2024-08-29 DOI: 10.1016/j.dci.2024.105258
Jim Kaufman

What are the future directions of the fields of developmental and comparative immunology? In thinking through this question as I write, I find myself marvelling at the very long ways that we have come since I began as a PhD student some 50 years ago. I think that we cannot know what technical and theoretical advances will emerge in the future, nor will our initial aims survive the realities of what appears in our sights, often from unexpected directions. I feel that we should not allow what we already know about some well-studied systems to blind us to the wide range of possibilities, and that remaining a humble seeker helps the uptake of new realities. Finally, it would be good to try answering the whole range of questions about developmental and comparative immunology, from what to how to why.

发育免疫学和比较免疫学领域的未来发展方向是什么?在写作过程中思考这个问题时,我发现自己惊叹于自 50 多年前我开始攻读博士学位以来,我们已经走过了漫长的道路。我认为,我们无法预知未来会出现什么样的技术和理论进步,我们最初的目标也无法经受住现实的考验,因为现实往往来自意想不到的方向。我认为,我们不应该让我们已经了解的一些经过深入研究的系统蒙蔽我们的双眼,使我们看不到广泛的可能性,保持谦虚的探索精神有助于接受新的现实。最后,我们不妨尝试回答有关发育免疫学和比较免疫学的所有问题,从 "是什么 "到 "如何做",再到 "为什么"。
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引用次数: 0
Preface to the special issue 特刊序言。
IF 2.7 3区 农林科学 Q1 FISHERIES Pub Date : 2024-08-29 DOI: 10.1016/j.dci.2024.105255
Geert F. Wiegertjes, Maria Forlenza, Christine A. Jansen
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引用次数: 0
Characterization of IgD and IgT with their expressional analysis following subtype II megalocytivirus vaccination and infection in rock bream (Oplegnathus fasciatus) 岩鲷(Oplegnathus fasciatus)接种亚型 II 巨细胞病毒疫苗和感染后 IgD 和 IgT 的特征及其表达分析。
IF 2.7 3区 农林科学 Q1 FISHERIES Pub Date : 2024-08-29 DOI: 10.1016/j.dci.2024.105248
Sungjae Ko, Suhee Hong

In this study, heavy chain genes of IgD and IgT were sequenced and characterized their gene expression in rock bream (Oplegnathus fasciatus). Rock bream (RB)-IgD cDNA is 3319 bp in length and encodes a leader region, variable domains, a μ1 domain, and seven constant domains (CH1–CH7). A membrane-bound (mIgT) and secretory form (sIgT) of RB-IgT cDNAs are 1902 bp and 1689 bp in length, respectively, and encode a leader region, variable domains, four constant domains (CH1–CH4) and C-terminus. Their predicted 3D-structure and phylogenetic relation were similar to those of other teleost. In healthy fish, RB-IgD and mIgT gene expressions were higher in major lymphoid organs and blood, while RB-sIgT gene was more highly expressed in midgut. IgT expressing cells were detected in melano-macrophage centers (MMC) of head kidney in immunohistochemistry analysis. Under immune stimulation in vitro, RB-IgD and IgT gene expressions were upregulated in head kidney and spleen cells by bovine serum albumin or a rock bream iridovirus (RBIV) vaccine. In vivo, their expressions were significantly upregulated in head kidney, blood, and gill upon vaccination. Especially, RB-mIgT gene expression in head kidney and blood was upregulated at day 3 after vaccination while upregulated at earlier time point of day 1 by challenge with RBIV. This may suggest that memory cells might be produced during the primary response by vaccination and rapidly proliferated by secondary immune response by viral infection. RB-sIgT gene expression was highly upregulated in peripheral blood in vaccinated fish after viral infection, indicating that IgT plays an important role in systemic immune response as well as mucosal immune system. Our findings provide information on the role of RB-IgT in adaptive immunity during vaccination and viral infection in the vaccinated fish.

本研究对岩鲷(Oplegnathus fasciatus)的 IgD 和 IgT 重链基因进行了测序,并确定了其基因表达的特征。岩鳊(RB)-IgD cDNA 长 3319 bp,编码一个领导区、可变结构域、μ1 结构域和七个恒定结构域(CH1-CH7)。膜结合型(mIgT)和分泌型(sIgT)的 RB-IgT cDNA 长度分别为 1902 bp 和 1689 bp,编码一个领导区、可变结构域、四个恒定结构域(CH1-CH4)和 C-末端。它们的预测三维结构和系统发育关系与其他远洋鱼类相似。在健康鱼类中,RB-IgD和mIgT基因在主要淋巴器官和血液中表达较高,而RB-sIgT基因在中肠中表达较高。免疫组化分析在头肾的黑色-巨噬细胞中心(MMC)检测到表达 IgT 的细胞。在体外免疫刺激下,牛血清白蛋白或鳊鱼虹彩病毒(RBIV)疫苗可上调头肾和脾脏细胞中 RB-IgD 和 IgT 基因的表达。在体内,接种疫苗后,它们在头部肾脏、血液和鳃中的表达明显上调。特别是在接种疫苗后第3天,RB-mIgT基因在头部肾脏和血液中的表达上调,而在接种RBIV疫苗后第1天的较早时间点,RB-mIgT基因的表达上调。这可能表明,记忆细胞可能是在疫苗接种的初级反应中产生,并在病毒感染的次级免疫反应中迅速增殖。病毒感染后,接种鱼外周血中的 RB-sIgT 基因表达高度上调,表明 IgT 在全身免疫反应和粘膜免疫系统中发挥着重要作用。我们的研究结果提供了接种疫苗和病毒感染期间 RB-IgT 在接种鱼适应性免疫中的作用。
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引用次数: 0
Invertebrate immunology – some thoughts about past and future research 无脊椎动物免疫学--对过去和未来研究的一些思考。
IF 2.7 3区 农林科学 Q1 FISHERIES Pub Date : 2024-08-28 DOI: 10.1016/j.dci.2024.105256
Kenneth Söderhäll
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引用次数: 0
TRIM103 activates the RLRs pathway to enhance antiviral response by targeting VP5 and VP7 TRIM103 可激活 RLRs 途径,通过靶向 VP5 和 VP7 增强抗病毒反应。
IF 2.7 3区 农林科学 Q1 FISHERIES Pub Date : 2024-08-28 DOI: 10.1016/j.dci.2024.105254
Beibei Qin , Zhao Lv , Hong Yang , Tiaoyi Xiao , Jianming Su

Grass carp (Ctenopharyngodon idella), crucial to global inland aquaculture with a production of 5.8 million tones in 2020, faces significant challenges from hemorrhagic disease caused by grass carp reovirus (GCRV). Rapid mutations compromise current vaccines, underscoring the need for a deeper understanding of antiviral mechanisms to enhance molecular marker-assisted selection. This study investigates the role of Tripartite Motif (TRIM) family in the innate immune response of grass carp, focusing on TRIM103 from Ctenopharyngodon Idella (CiTRIM103), a member of the TRIM-B30.2 family, which includes proteins with the B30.2 domain at the N-terminus, known for antiviral properties in teleosts. CiTRIM103 bind to the outer coat proteins VP5 and VP7 of GCRV. This binding is theorized to strengthen the function of the RIG-I-like Receptor (RLR) signaling pathway, crucial for antiviral responses. Demonstrations using overexpression and RNA interference (RNAi) techniques have shown that CiTRIM103 effectively inhibits GCRV replication. Moreover, molecular docking and pulldown assays suggest potential binding interactions of CiTRIM103's B30.2 domain with GCRV outer coat proteins VP5 and VP7. These interactions impede viral replication, enhance RLR receptor expression, and activate key transcription factors to induce type I interferons (IFNs). These findings elucidate the antiviral mechanisms of CiTRIM103, provide a foundation for future Molecular genetic breeding in grass carp.

草鱼(Ctenopharyngodon idella)对全球内陆水产养殖业至关重要,2020 年的产量将达到 580 万吨,但它却面临着草鱼再病毒(GCRV)引起的出血性疾病的巨大挑战。快速突变损害了现有疫苗,突出表明需要深入了解抗病毒机制,以加强分子标记辅助选择。本研究调查了三方动因(TRIM)家族在草鱼先天性免疫反应中的作用,重点研究了Ctenopharyngodon Idella的TRIM103(CiTRIM103),它是TRIM-B30.2家族的成员,该家族包括N端具有B30.2结构域的蛋白质,在远缘动物中具有抗病毒特性。CiTRIM103 与 GCRV 的外衣蛋白 VP5 和 VP7 结合。据推测,这种结合能加强 RIG-I-like Receptor(RLR)信号通路的功能,而 RIG-I-like Receptor 对抗病毒反应至关重要。利用过表达和 RNA 干扰(RNAi)技术进行的实验表明,CiTRIM103 能有效抑制 GCRV 的复制。此外,分子对接和下拉试验表明,CiTRIM103 的 B30.2 结构域与 GCRV 外衣蛋白 VP5 和 VP7 有潜在的结合相互作用。这些相互作用阻碍了病毒的复制,增强了 RLR 受体的表达,并激活了关键转录因子以诱导 I 型干扰素(IFNs)。这些发现阐明了 CiTRIM103 的抗病毒机制,为未来草鱼分子遗传育种奠定了基础。
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引用次数: 0
期刊
Developmental and comparative immunology
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