One of the most highly induced genes in zebrafish (Danio rerio) larvae after infection with the nodavirus red-spotted grouper nervous necrosis virus (RGNNV) was a member of the immunoglobulin superfamily (IgSF), which has remained uncharacterized and erroneously annotated in zebrafish and other fish species, such as galectin 17 (lgals17). We characterized this gene and named it immunoglobulin (Ig)-like domain-containing protein (igldcp), a new member of the IgSF that does not possess orthologs in mammals. Igldcp expression is induced by viral infection and it belongs to the group of interferon-stimulated genes (ISGs). In vitro overexpression of igldcp decreased RGNNV replication, whereas in vivo knockdown of this gene had the opposite effect, resulting in increased larval mortality. RNA-Seq analyses of larvae overexpressing igldcp in the absence or presence of infection with RGNNV showed that the main processes affected by Igldcp could be directly involved in the regulation of various cellular processes associated with the modulation of the immune system.
{"title":"Characterization of a fish-specific Immunoglobulin-like domain-containing protein (Igldcp) in zebrafish (Danio rerio) induced after nodavirus infection.","authors":"Nieves Martínez-López, Patricia Pereiro, Amaro Saco, Raquel Lama, Antonio Figueras, Beatriz Novoa","doi":"10.1016/j.dci.2024.105285","DOIUrl":"https://doi.org/10.1016/j.dci.2024.105285","url":null,"abstract":"<p><p>One of the most highly induced genes in zebrafish (Danio rerio) larvae after infection with the nodavirus red-spotted grouper nervous necrosis virus (RGNNV) was a member of the immunoglobulin superfamily (IgSF), which has remained uncharacterized and erroneously annotated in zebrafish and other fish species, such as galectin 17 (lgals17). We characterized this gene and named it immunoglobulin (Ig)-like domain-containing protein (igldcp), a new member of the IgSF that does not possess orthologs in mammals. Igldcp expression is induced by viral infection and it belongs to the group of interferon-stimulated genes (ISGs). In vitro overexpression of igldcp decreased RGNNV replication, whereas in vivo knockdown of this gene had the opposite effect, resulting in increased larval mortality. RNA-Seq analyses of larvae overexpressing igldcp in the absence or presence of infection with RGNNV showed that the main processes affected by Igldcp could be directly involved in the regulation of various cellular processes associated with the modulation of the immune system.</p>","PeriodicalId":11228,"journal":{"name":"Developmental and comparative immunology","volume":null,"pages":null},"PeriodicalIF":2.7,"publicationDate":"2024-11-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142603423","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-11-02DOI: 10.1016/j.dci.2024.105284
Xue Kong , Wei Wang , Sunan Xia , Ying Zhi , Yuefeng Cai , Haibin Zhang , Xin Shen
Within cold seep environments, the Vesicomyidae clam emerges as a prevalent species, distinguished by its symbiotic relationship with microorganisms housed within its organ gill. Given the extreme conditions and the symbiotic nature of this association, investigating the host's immune genes, particularly immune recognition receptors, is essential for understanding their role in facilitating host-symbiotic interactions. Three short peptidoglycan recognition proteins (PGRPs) were identified in the clam. AmPGRP-S1, -S2, and -S3 were found to possess conserved amidase binding sites and Zn2+ binding sites. Quantitative Real-time PCR (qRT-PCR) analysis revealed differential expression patterns among the PGRPs. AmPGRP-S1 and AmPGRP-S2 exhibited elevated expression levels in the gill, while AmPGRP-S3 displayed the highest expression in the adductor muscle. Functional experiments demonstrated that recombinant AmPGRP-S1, -S2, and -S3 (rAmPGRPs) exhibited binding capabilities to both L-PGN and D-PGN (peptidoglycan). Notably, rAmPGRP-S1 and -S2 possessed Zn2+-independent amidase activity, while rAmPGRP-S3 lacked this enzymatic function. rAmPGRPs were shown to bind to five different bacterial species. Among these, rAmPGRP-S1 inhibited Escherichia coli and Bacillus subtilis, while rAmPGRP-S2 and -S3 inhibited Bacillus subtilis in the absence of Zn2+. In the presence of Zn2+, rAmPGRP-S1 and -S2 exhibited enhanced inhibitory activity against Staphylococcus aureus or Bacillus subtilis. These findings suggest that AmPGRPs may play a pivotal role in mediating the interaction between the host and endosymbiotic bacteria, functioning as PGN and microbe receptors, antibacterial effectors, and regulators of host-microbe symbiosis. These results contribute to our understanding of the adaptive mechanisms of deep-sea organisms to the challenging cold seep environments.
{"title":"Molecular and functional characterization of short peptidoglycan recognition proteins in Vesicomyidae clam","authors":"Xue Kong , Wei Wang , Sunan Xia , Ying Zhi , Yuefeng Cai , Haibin Zhang , Xin Shen","doi":"10.1016/j.dci.2024.105284","DOIUrl":"10.1016/j.dci.2024.105284","url":null,"abstract":"<div><div>Within cold seep environments, the Vesicomyidae clam emerges as a prevalent species, distinguished by its symbiotic relationship with microorganisms housed within its organ gill. Given the extreme conditions and the symbiotic nature of this association, investigating the host's immune genes, particularly immune recognition receptors, is essential for understanding their role in facilitating host-symbiotic interactions. Three short peptidoglycan recognition proteins (PGRPs) were identified in the clam. AmPGRP-S1, -S2, and -S3 were found to possess conserved amidase binding sites and Zn<sup>2+</sup> binding sites. Quantitative Real-time PCR (qRT-PCR) analysis revealed differential expression patterns among the PGRPs. AmPGRP-S1 and AmPGRP-S2 exhibited elevated expression levels in the gill, while AmPGRP-S3 displayed the highest expression in the adductor muscle. Functional experiments demonstrated that recombinant AmPGRP-S1, -S2, and -S3 (rAmPGRPs) exhibited binding capabilities to both L-PGN and D-PGN (peptidoglycan). Notably, rAmPGRP-S1 and -S2 possessed Zn<sup>2+</sup>-independent amidase activity, while rAmPGRP-S3 lacked this enzymatic function. rAmPGRPs were shown to bind to five different bacterial species. Among these, rAmPGRP-S1 inhibited <em>Escherichia coli</em> and <em>Bacillus subtilis</em>, while rAmPGRP-S2 and -S3 inhibited <em>Bacillus subtilis</em> in the absence of Zn<sup>2<em>+</em></sup>. In the presence of Zn<sup>2+</sup>, rAmPGRP-S1 and -S2 exhibited enhanced inhibitory activity against <em>Staphylococcus aureus</em> or <em>Bacillus subtilis</em>. These findings suggest that AmPGRPs may play a pivotal role in mediating the interaction between the host and endosymbiotic bacteria, functioning as PGN and microbe receptors, antibacterial effectors, and regulators of host-microbe symbiosis. These results contribute to our understanding of the adaptive mechanisms of deep-sea organisms to the challenging cold seep environments.</div></div>","PeriodicalId":11228,"journal":{"name":"Developmental and comparative immunology","volume":null,"pages":null},"PeriodicalIF":2.7,"publicationDate":"2024-11-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142567814","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Membrane-associated Ring-CH 5 (MARCH5) is a mitochondrial E3 ubiquitin ligase playing a key role in the regulation of mitochondrial dynamics. In mammals, MARCH5 negatively regulates mitochondrial antiviral signaling (MAVS) protein aggregation during viral infection and hampers downstream type I interferon signaling to prevent excessive immune activation. However, its precise functional role in the teleost immune system remains unclear. This study investigated the molecular characteristics and immune response of the MARCH5 ortholog in Amphiprion clarkii (A. clarkii; AcMARCH5). The predicted AcMARCH5 protein sequence consists of 287 amino acids with a molecular weight of 32.02 kDa and a theoretical isoelectric point of 9.11. It contains four C-terminal transmembrane (TM) domains and an N-terminal RING cysteine-histidine (CH) domain, which directly regulates ubiquitin transfer. Multiple sequence alignment revealed a high level of conservation between AcMARCH5 and its orthologs in other vertebrate species. Under normal physiological conditions, AcMARCH5 showed the highest mRNA expression in the muscle, brain, and kidney tissues of A. clarkii. Upon stimulation with polyinosinic:polycytidylic acid (Poly I:C), lipopolysaccharide (LPS), and Vibrio harveyi, AcMARCH5 expression was drastically modulated. Functional assays showed that overexpression of AcMARCH5 in fathead minnow (FHM) cells downregulated antiviral gene expression, accompanied by enhanced viral hemorrhagic septicemia virus (VHSV) replication. In murine macrophages, AcMARCH5 overexpression markedly reduced the production of pro-inflammatory cytokines in response to poly I:C treatment. Additionally, AcMARCH5 exhibited an anti-apoptotic effect in H2O2-treated FHM cells. Collectively, these results suggest that AcMARCH5 may play a role in maintaining cellular homeostasis under disease and stress conditions in A. clarkii.
膜相关环-CH 5（MARCH5）是一种线粒体 E3 泛素连接酶，在线粒体动力学调控中发挥着关键作用。在哺乳动物中，MARCH5 在病毒感染期间负向调节线粒体抗病毒信号转导（MAVS）蛋白的聚集，并阻碍下游 I 型干扰素信号转导，以防止过度的免疫激活。然而，它在远洋鱼类免疫系统中的确切功能作用仍不清楚。本研究调查了克氏栉水母（Amphiprion clarkii；AcMARCH5）MARCH5直向同源物的分子特征和免疫反应。预测的 AcMARCH5 蛋白序列由 287 个氨基酸组成，分子量为 32.02 kDa，理论等电点为 9.11。它包含四个 C 端跨膜（TM）结构域和一个 N 端 RING 半胱氨酸-组氨酸（CH）结构域，后者直接调节泛素的转移。多重序列比对显示，AcMARCH5与其在其他脊椎动物中的同源物之间存在高度保守性。在正常生理条件下，AcMARCH5 在克氏原鲤的肌肉、脑和肾组织中的 mRNA 表达量最高。在多聚肌苷酸：多聚胞苷酸（Poly I:C）、脂多糖（LPS）和哈维弧菌的刺激下，AcMARCH5的表达受到了极大的影响。功能测试显示，在黑头鲦鱼（FHM）细胞中过表达 AcMARCH5 会降低抗病毒基因的表达，同时增强病毒性出血性败血症病毒（VHSV）的复制。在小鼠巨噬细胞中，AcMARCH5 的过表达明显减少了聚 I:C 处理后促炎细胞因子的产生。此外，AcMARCH5 在 H2O2 处理的 FHM 细胞中表现出抗凋亡作用。总之，这些结果表明，AcMARCH5 可能在 A. clarkii 疾病和应激条件下维持细胞稳态方面发挥作用。
{"title":"Molecular depiction and functional delineation of E3 ubiquitin ligase MARCH5 in yellowtail clownfish (Amphiprion clarkii)","authors":"B.P.M. Vileka Jayamali , H.M.S.M. Wijerathna , D.M.K.P. Sirisena , H.A.C.R. Hanchapola , W.A.D.L.R. Warnakula , U.P.E. Arachchi , D.S. Liyanage , Sumi Jung , Qiang Wan , Jehee Lee","doi":"10.1016/j.dci.2024.105283","DOIUrl":"10.1016/j.dci.2024.105283","url":null,"abstract":"<div><div>Membrane-associated Ring-CH 5 (MARCH5) is a mitochondrial E3 ubiquitin ligase playing a key role in the regulation of mitochondrial dynamics. In mammals, MARCH5 negatively regulates mitochondrial antiviral signaling (MAVS) protein aggregation during viral infection and hampers downstream type I interferon signaling to prevent excessive immune activation. However, its precise functional role in the teleost immune system remains unclear. This study investigated the molecular characteristics and immune response of the MARCH5 ortholog in <em>Amphiprion clarkii</em> (<em>A. clarkii</em>; AcMARCH5). The predicted AcMARCH5 protein sequence consists of 287 amino acids with a molecular weight of 32.02 kDa and a theoretical isoelectric point of 9.11. It contains four C-terminal transmembrane (TM) domains and an N-terminal RING cysteine-histidine (CH) domain, which directly regulates ubiquitin transfer. Multiple sequence alignment revealed a high level of conservation between AcMARCH5 and its orthologs in other vertebrate species. Under normal physiological conditions, <em>AcMARCH5</em> showed the highest mRNA expression in the muscle, brain, and kidney tissues of <em>A. clarkii</em>. Upon stimulation with polyinosinic:polycytidylic acid (Poly I:C), lipopolysaccharide (LPS), and <em>Vibrio harveyi</em>, <em>AcMARCH5</em> expression was drastically modulated. Functional assays showed that overexpression of AcMARCH5 in fathead minnow (FHM) cells downregulated antiviral gene expression, accompanied by enhanced viral hemorrhagic septicemia virus (VHSV) replication. In murine macrophages, AcMARCH5 overexpression markedly reduced the production of pro-inflammatory cytokines in response to poly I:C treatment. Additionally, AcMARCH5 exhibited an anti-apoptotic effect in H<sub>2</sub>O<sub>2</sub>-treated FHM cells. Collectively, these results suggest that AcMARCH5 may play a role in maintaining cellular homeostasis under disease and stress conditions in <em>A. clarkii</em>.</div></div>","PeriodicalId":11228,"journal":{"name":"Developmental and comparative immunology","volume":null,"pages":null},"PeriodicalIF":2.7,"publicationDate":"2024-10-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142557392","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-10-20DOI: 10.1016/j.dci.2024.105282
Carolina Salazar , Nicolás Ojeda , Luis Mercado
Endocrine Disruptor Compounds (EDCs) in the aquatic environment have acquired pronounced relevance due to their toxic effect on the aquatic flora and fauna. Xenoestrogens are EDCs that possess estrogenic activity and, thus, disrupt normal estrogen signaling, affecting different functions, such as immune system processes. Two relevant xenoestrogens discarded into fresh and seawater are 4-nonylphenol (NP) and 17⍺-Ethynyl Estradiol (EE2). Considering that the piscicultures of Salmo salar can be located at sites of potential exposure to xenoestrogen-containing effluxes, it is crucial to understand the effect of xenoestrogens on the immune response and its possible molecular mechanism in this species. Our studies reveal an increase in the expression of the receptor era and erb at early times of exposure, a disrupted expression of pro-inflammatory cytokines (il1b and tnfa), an upregulation of ssa-miR-146a-5p, ssa-miR-125 b-5p, and downregulation of ssa-miR-145–5p in ASK cells exposed to estrogen and xenoestrogen, could potentially lead to new strategies for mitigating the effects of xenoestrogens on Salmo salar immune response.
{"title":"Dysregulated proinflammatory cytokines and immune-related miRNAs in ASK cells exposed to 17⍺-Ethynyl estradiol and 4-nonylphenol","authors":"Carolina Salazar , Nicolás Ojeda , Luis Mercado","doi":"10.1016/j.dci.2024.105282","DOIUrl":"10.1016/j.dci.2024.105282","url":null,"abstract":"<div><div>Endocrine Disruptor Compounds (EDCs) in the aquatic environment have acquired pronounced relevance due to their toxic effect on the aquatic flora and fauna. Xenoestrogens are EDCs that possess estrogenic activity and, thus, disrupt normal estrogen signaling, affecting different functions, such as immune system processes. Two relevant xenoestrogens discarded into fresh and seawater are 4-nonylphenol (NP) and 17⍺-Ethynyl Estradiol (EE2). Considering that the piscicultures of <em>Salmo salar</em> can be located at sites of potential exposure to xenoestrogen-containing effluxes, it is crucial to understand the effect of xenoestrogens on the immune response and its possible molecular mechanism in this species. Our studies reveal an increase in the expression of the receptor <em>era</em> and <em>erb</em> at early times of exposure, a disrupted expression of pro-inflammatory cytokines (<em>il1b</em> and <em>tnfa</em>), an upregulation of ssa-miR-146a-5p, ssa-miR-125 b-5p, and downregulation of ssa-miR-145–5p in ASK cells exposed to estrogen and xenoestrogen, could potentially lead to new strategies for mitigating the effects of xenoestrogens on <em>Salmo salar</em> immune response.</div></div>","PeriodicalId":11228,"journal":{"name":"Developmental and comparative immunology","volume":null,"pages":null},"PeriodicalIF":2.7,"publicationDate":"2024-10-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142496903","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-10-18DOI: 10.1016/j.dci.2024.105281
Dong-li Li , Wen-lin Wu , Hai-peng Liu
White spot syndrome virus (WSSV) is a large nuclear-replicating DNA virus of crustaceans such as shrimp and crayfish; however, the molecular mechanisms facilitating its transport from the invasion site to the cell nucleus have not yet been well elucidated. In this study, a CqProfilin (CqPFN) with a conserved PROF domain was identified from the red claw crayfish Cherax quadricarinatus. CqPFN was ubiquitously expressed in all examined tissues and hemocyte, with the highest levels in the hemocyte, followed by hematopoietic tissue (Hpt) from which the hemocyte were derived in crayfish. The transcript of WSSV genes such as IE1 and VP28 was obviously decreased both in vivo in hemocyte and Hpt, as well as in vitro in cultured Hpt cells, after CqPFN gene silencing; in contrast, the expression of viral genes was significantly increased by the introduction of a recombinant CqPFN protein in Hpt cells in vitro. Moreover, CqPFN was clearly colocalized with the main viral nucleocapsid protein VP664 and F-actin cytoskeleton, respectively, during the early stage of WSSV infection in Hpt cells. In addition, CqPFN was confirmed to interact with a truncated VP6642,405-2,535 and another viral nucleocapsid protein VP15 of WSSV and Cqβ-Actin from Hpt by co-immunoprecipitation assays. Further studies found that VP664 also colocalized with F-actin in the Hpt cell cytoplasm after WSSV infection, suggesting that the actin cytoskeleton was involved in the intracellular transport of incoming viral nucleocapsid. Taken together, CqPFN might combine with the actin cytoskeleton to promote WSSV infection through binding with viral nucleocapsid proteins VP664 and VP15, promoting intracellular transport of viral incoming nucleocapsid for further releasing genome into the nucleus for transcription. Collectively, these results provided an understanding of the WSSV pathogenesis, which will contribute to the development of an antiviral strategy against WSSV disease.
{"title":"CqProfilin enhances WSSV infection by promoting viral intracellular transport through binding to both viral nucleocapsid and actin cytoskeleton","authors":"Dong-li Li , Wen-lin Wu , Hai-peng Liu","doi":"10.1016/j.dci.2024.105281","DOIUrl":"10.1016/j.dci.2024.105281","url":null,"abstract":"<div><div>White spot syndrome virus (WSSV) is a large nuclear-replicating DNA virus of crustaceans such as shrimp and crayfish; however, the molecular mechanisms facilitating its transport from the invasion site to the cell nucleus have not yet been well elucidated. In this study, a <em>Cq</em>Profilin (<em>Cq</em>PFN) with a conserved PROF domain was identified from the red claw crayfish <em>Cherax quadricarinatus</em>. <em>CqPFN</em> was ubiquitously expressed in all examined tissues and hemocyte, with the highest levels in the hemocyte, followed by hematopoietic tissue (Hpt) from which the hemocyte were derived in crayfish. The transcript of WSSV genes such as <em>IE1</em> and <em>VP28</em> was obviously decreased both <em>in vivo</em> in hemocyte and Hpt, as well as <em>in vitro</em> in cultured Hpt cells, after <em>CqPFN</em> gene silencing; in contrast, the expression of viral genes was significantly increased by the introduction of a recombinant <em>Cq</em>PFN protein in Hpt cells <em>in vitro</em>. Moreover, <em>Cq</em>PFN was clearly colocalized with the main viral nucleocapsid protein VP664 and F-actin cytoskeleton, respectively, during the early stage of WSSV infection in Hpt cells. In addition, <em>Cq</em>PFN was confirmed to interact with a truncated VP664<sup>2,405-2,535</sup> and another viral nucleocapsid protein VP15 of WSSV and <em>Cq</em>β-Actin from Hpt by co-immunoprecipitation assays. Further studies found that VP664 also colocalized with F-actin in the Hpt cell cytoplasm after WSSV infection, suggesting that the actin cytoskeleton was involved in the intracellular transport of incoming viral nucleocapsid. Taken together, <em>Cq</em>PFN might combine with the actin cytoskeleton to promote WSSV infection through binding with viral nucleocapsid proteins VP664 and VP15, promoting intracellular transport of viral incoming nucleocapsid for further releasing genome into the nucleus for transcription. Collectively, these results provided an understanding of the WSSV pathogenesis, which will contribute to the development of an antiviral strategy against WSSV disease.</div></div>","PeriodicalId":11228,"journal":{"name":"Developmental and comparative immunology","volume":null,"pages":null},"PeriodicalIF":2.7,"publicationDate":"2024-10-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142460364","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-10-11DOI: 10.1016/j.dci.2024.105280
Sara Pedrazzoli, Giulia Graziosi, Roberta Salaroli, Elena Catelli, Caterina Lupini
Infectious bursal disease virus (IBDV) is a significant pathogen in poultry, causing acute immunosuppressive disease in young chickens. While B-lymphocyte involvement in IBDV pathogenesis is known, the role of T-cells is incompletely understood. This systematic review presents the alterations in chicken T-lymphocyte subsets after IBDV exposure, assessed by flow cytometry analysis. Four databases were queried for identifying eligible studies focused on experimental infections measuring T-lymphocyte changes in the bursa of Fabricius, spleen, thymus, and peripheral blood mononuclear cells. Of 488 studies found, 25 met the pre-established criteria and were included in the qualitative synthesis of results. Most studies analysed T-lymphocyte responses during the acute phase of IBDV infection, primarily focusing on CD4+ and CD8+ T-cells. Other subsets, such as γδ T-cells and double-positive CD4+CD8+ T-cells, were less frequently investigated. An increase in T-lymphocytes was noted in the bursa of Fabricius, suggesting their active role in viral clearance. In the spleen, CD4+ T-cells commonly increased, while CD8+ responses varied among studies. Increased levels in T-cells were also noted during the chronic infection in the bursa of Fabricius, possibly due to persistent viral antigens. Overall, variations in flow cytometry methods and T-cell output reporting were noted among studies.
Based on the data collected, further investigation into diverse T-cell subpopulations beyond CD4+ and CD8+ is needed, as well as the standardization of flow cytometry assays in chickens.
传染性法氏囊病病毒（IBDV）是家禽的一种重要病原体，可导致幼鸡急性免疫抑制性疾病。虽然已知 B 淋巴细胞参与了 IBDV 的发病过程，但对 T 细胞的作用还不完全了解。本系统综述通过流式细胞术分析评估了鸡暴露于 IBDV 后 T 淋巴细胞亚群的变化。我们查询了四个数据库，以确定符合条件的研究，这些研究侧重于测量法氏囊、脾脏、胸腺和外周血单核细胞中 T 淋巴细胞的变化。在找到的 488 项研究中，有 25 项符合预先设定的标准，并纳入了定性结果综述。大多数研究分析了 IBDV 感染急性期的 T 淋巴细胞反应，主要侧重于 CD4+ 和 CD8+ T 细胞。对其他亚群，如 γδ T 细胞和 CD4+CD8+ 双阳性 T 细胞的研究较少。法氏囊中的 T 淋巴细胞有所增加，表明它们在清除病毒方面发挥了积极作用。在脾脏，CD4+ T 细胞普遍增加，而 CD8+ 的反应则因研究而异。在法氏囊慢性感染期间，T 细胞水平也有所提高，这可能是由于病毒抗原的持续存在。总体而言，不同研究的流式细胞术方法和 T 细胞输出报告存在差异。根据收集到的数据，需要进一步研究 CD4+ 和 CD8+ 以外的各种 T 细胞亚群，并对鸡的流式细胞术检测方法进行标准化。
{"title":"Dynamic alterations in T-lymphocyte subsets assessed by flow cytometry in chickens following exposure to infectious bursal disease virus: A systematic review","authors":"Sara Pedrazzoli, Giulia Graziosi, Roberta Salaroli, Elena Catelli, Caterina Lupini","doi":"10.1016/j.dci.2024.105280","DOIUrl":"10.1016/j.dci.2024.105280","url":null,"abstract":"<div><div>Infectious bursal disease virus (IBDV) is a significant pathogen in poultry, causing acute immunosuppressive disease in young chickens. While B-lymphocyte involvement in IBDV pathogenesis is known, the role of T-cells is incompletely understood. This systematic review presents the alterations in chicken T-lymphocyte subsets after IBDV exposure, assessed by flow cytometry analysis. Four databases were queried for identifying eligible studies focused on experimental infections measuring T-lymphocyte changes in the bursa of Fabricius, spleen, thymus, and peripheral blood mononuclear cells. Of 488 studies found, 25 met the pre-established criteria and were included in the qualitative synthesis of results. Most studies analysed T-lymphocyte responses during the acute phase of IBDV infection, primarily focusing on CD4<sup>+</sup> and CD8<sup>+</sup> T-cells. Other subsets, such as γδ T-cells and double-positive CD4<sup>+</sup>CD8<sup>+</sup> T-cells, were less frequently investigated. An increase in T-lymphocytes was noted in the bursa of Fabricius, suggesting their active role in viral clearance. In the spleen, CD4<sup>+</sup> T-cells commonly increased, while CD8<sup>+</sup> responses varied among studies. Increased levels in T-cells were also noted during the chronic infection in the bursa of Fabricius, possibly due to persistent viral antigens. Overall, variations in flow cytometry methods and T-cell output reporting were noted among studies.</div><div>Based on the data collected, further investigation into diverse T-cell subpopulations beyond CD4<sup>+</sup> and CD8<sup>+</sup> is needed, as well as the standardization of flow cytometry assays in chickens.</div></div>","PeriodicalId":11228,"journal":{"name":"Developmental and comparative immunology","volume":null,"pages":null},"PeriodicalIF":2.7,"publicationDate":"2024-10-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142460365","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-10-11DOI: 10.1016/j.dci.2024.105279
Thi Hao Vu , Chaeeun Kim , Anh Duc Truong , Hyun S. Lillehoj , Yeong Ho Hong
This study describes the first successful cloning and functional characterization of chicken CX3CL1, a chemokine involved in immune cell migration and inflammatory responses. Evolutionary analyses revealed its close relation to CX3CL1 from other avian species, particularly duck, turkey, and quail. Structurally, chicken CX3CL1 includes a signal peptide and a chemokine interleukin-8-like domain characterized by unique alpha-helices and disulfide bonds. Additionally, we produced and purified recombinant CX3CL1 protein and assessed its endotoxin levels. Chemotaxis assays revealed that CX3CL1 significantly enhances the migration of HD11 macrophages and CU91 T cells. Furthermore, recombinant CX3CL1 induced the expression of pro-inflammatory cytokines (TNF-α, IFN-β, IFN-γ, IL-6, and CCL20) in a time-dependent manner, while exerting differential effects on anti-inflammatory cytokines (IL-4, IL-10). Conversely, transfection with siCX3CL1 or siCX3CR1 led to the downregulation of these responses. We also observed activation of the MAPK, NF-κB, and JAK/STAT pathways, evidenced by increased phosphorylation of key signaling molecules. These findings underscore the crucial role of chicken CX3CL1 in regulating immune responses, cell migration, and the activation of key signaling pathways. This study provides valuable insights into the immunomodulatory functions of soluble CX3CL1, highlighting its potential as a therapeutic target for inflammatory conditions and enhancing our understanding of immune cell dynamics.
{"title":"Unveiling the immunomodulatory role of soluble chicken fractalkine: Insights from functional characterization and pathway activation analyses","authors":"Thi Hao Vu , Chaeeun Kim , Anh Duc Truong , Hyun S. Lillehoj , Yeong Ho Hong","doi":"10.1016/j.dci.2024.105279","DOIUrl":"10.1016/j.dci.2024.105279","url":null,"abstract":"<div><div>This study describes the first successful cloning and functional characterization of chicken CX3CL1, a chemokine involved in immune cell migration and inflammatory responses. Evolutionary analyses revealed its close relation to CX3CL1 from other avian species, particularly duck, turkey, and quail. Structurally, chicken CX3CL1 includes a signal peptide and a chemokine interleukin-8-like domain characterized by unique alpha-helices and disulfide bonds. Additionally, we produced and purified recombinant CX3CL1 protein and assessed its endotoxin levels. Chemotaxis assays revealed that CX3CL1 significantly enhances the migration of HD11 macrophages and CU91 T cells. Furthermore, recombinant CX3CL1 induced the expression of pro-inflammatory cytokines (TNF-α, IFN-β, IFN-γ, IL-6, and CCL20) in a time-dependent manner, while exerting differential effects on anti-inflammatory cytokines (IL-4, IL-10). Conversely, transfection with siCX3CL1 or siCX3CR1 led to the downregulation of these responses. We also observed activation of the MAPK, NF-κB, and JAK/STAT pathways, evidenced by increased phosphorylation of key signaling molecules. These findings underscore the crucial role of chicken CX3CL1 in regulating immune responses, cell migration, and the activation of key signaling pathways. This study provides valuable insights into the immunomodulatory functions of soluble CX3CL1, highlighting its potential as a therapeutic target for inflammatory conditions and enhancing our understanding of immune cell dynamics.</div></div>","PeriodicalId":11228,"journal":{"name":"Developmental and comparative immunology","volume":null,"pages":null},"PeriodicalIF":2.7,"publicationDate":"2024-10-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142444729","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-10-10DOI: 10.1016/j.dci.2024.105278
Yuexuan Wang , Yewen Wang , Yunxiang Jiang , Qiwei Qin , Shina Wei
Cathepsin X, a class of cysteine proteases in the lysosome, involved in intracellular protein degradation processes. Numerous reports revealed that many kinds of cysteine proteases played a crucial role in pathogen invasion. To investigate the relationship between cathepsin X of teleost fish and virus infection, EcCX was cloned and characterized in the orange-spotted grouper, Epinephelus coioides. The open reading frame (ORF) of EcCX included 909 nucleotides and encoded a protein consisting of 302 amino acids, which shared 75% and 56% identity with zebrafish and humans, respectively. The protein EcCX mainly consisted of a signal peptide (1–19 aa), a pro-pre-peptide region (20–55 aa), and a mature cysteine protease region (56–302 aa). Subcellular localization analysis showed that EcCX was mainly distributed in the cytoplasm, but EcCX ectoped to the vicinity of apoptotic vesicles in FHM cells during SGIV infection. Following stimulation with SGIV or Poly (dA:dT), there was a notable rise in the expression levels of EcCX. EcCX overexpression facilitated virus infection, upregulated the production of inflammatory factors, and induced the activation of the NF-κB promoter. Furthermore, the overexpression of EcCX also accelerated the process of SGIV-induced apoptosis, potentially by enhancing the promoter activity of P53 and AP-1. Overall, our findings demonstrated a correlation between the function of EcCX and SGIV infection, providing a new understanding of the mechanisms involved in fish virus infection.
{"title":"The essential function of cathepsin X of the orange-spotted grouper, Epinephelus coioides during SGIV infection","authors":"Yuexuan Wang , Yewen Wang , Yunxiang Jiang , Qiwei Qin , Shina Wei","doi":"10.1016/j.dci.2024.105278","DOIUrl":"10.1016/j.dci.2024.105278","url":null,"abstract":"<div><div>Cathepsin X, a class of cysteine proteases in the lysosome, involved in intracellular protein degradation processes. Numerous reports revealed that many kinds of cysteine proteases played a crucial role in pathogen invasion. To investigate the relationship between cathepsin X of teleost fish and virus infection, EcCX was cloned and characterized in the orange-spotted grouper, <em>Epinephelus coioides</em>. The open reading frame (ORF) of EcCX included 909 nucleotides and encoded a protein consisting of 302 amino acids, which shared 75% and 56% identity with zebrafish and humans, respectively. The protein EcCX mainly consisted of a signal peptide (1–19 aa), a pro-pre-peptide region (20–55 aa), and a mature cysteine protease region (56–302 aa). Subcellular localization analysis showed that EcCX was mainly distributed in the cytoplasm, but EcCX ectoped to the vicinity of apoptotic vesicles in FHM cells during SGIV infection. Following stimulation with SGIV or Poly (dA:dT), there was a notable rise in the expression levels of EcCX. EcCX overexpression facilitated virus infection, upregulated the production of inflammatory factors, and induced the activation of the NF-κB promoter. Furthermore, the overexpression of EcCX also accelerated the process of SGIV-induced apoptosis, potentially by enhancing the promoter activity of P53 and AP-1. Overall, our findings demonstrated a correlation between the function of EcCX and SGIV infection, providing a new understanding of the mechanisms involved in fish virus infection.</div></div>","PeriodicalId":11228,"journal":{"name":"Developmental and comparative immunology","volume":null,"pages":null},"PeriodicalIF":2.7,"publicationDate":"2024-10-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142444730","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Copepods are small crustaceans that live in microorganism-rich aquatic environments and provide a key supply of live food for fish and shellfish larviculture. To better understand the host-pathogen interaction between the copepod and Vibrio parahaemolyticus causing acute hepatopancreatic necrosis disease (VPAHPND), the comparative transcriptome and microbiome analyses were conducted in copepod Apocyclops royi-TH following VPAHPND infection. Transcriptome analysis identified a total of 836 differentially expressed genes, with 275 upregulated and 561 downregulated genes. Subsequent analysis showed that a total of 37 differentially expressed genes were associated with the innate immune system, including 16 upregulated genes related to Toll-like receptor signaling pathway, antimicrobial peptides, and stress response genes, and 21 downregulated genes associated with immunological modulators, signaling molecules, and apoptosis-related proteins. Analysis of the copepod microbiome following VPAHPND infection showed that the microbes changed significantly after bacterial infection, with a reduced alpha diversity accompanied by the increased level of Proteobacteria and decreased levels of Bdellovibrionota, Bacteroidota, and Verrucomicrobiota. The population of Vibrio genera were increased significantly, while several other genera, including Denitromonas, Nitrosomonas, Blastopirellula, Fusibacter, Alteromonas, KI89A_clade, and Ruegeria, were decreased significantly after infection. These findings suggest that VPAHPND infection has a significant impact on the immune defense and the composition of the copepod microbiota.
{"title":"Transcriptomic and microbiome analyses of copepod Apocyclops royi in response to an AHPND-causing strain of Vibrio parahaemolyticus","authors":"Natkanokporn Prayoonmaneerat , Walaiporn Charoensapsri , Piti Amparyup , Chanprapa Imjongjirak","doi":"10.1016/j.dci.2024.105277","DOIUrl":"10.1016/j.dci.2024.105277","url":null,"abstract":"<div><div>Copepods are small crustaceans that live in microorganism-rich aquatic environments and provide a key supply of live food for fish and shellfish larviculture. To better understand the host-pathogen interaction between the copepod and <em>Vibrio parahaemolyticus</em> causing acute hepatopancreatic necrosis disease (VP<sub>AHPND</sub>), the comparative transcriptome and microbiome analyses were conducted in copepod <em>Apocyclops royi</em>-TH following VP<sub>AHPND</sub> infection. Transcriptome analysis identified a total of 836 differentially expressed genes, with 275 upregulated and 561 downregulated genes. Subsequent analysis showed that a total of 37 differentially expressed genes were associated with the innate immune system, including 16 upregulated genes related to Toll-like receptor signaling pathway, antimicrobial peptides, and stress response genes, and 21 downregulated genes associated with immunological modulators, signaling molecules, and apoptosis-related proteins. Analysis of the copepod microbiome following VP<sub>AHPND</sub> infection showed that the microbes changed significantly after bacterial infection, with a reduced alpha diversity accompanied by the increased level of Proteobacteria and decreased levels of Bdellovibrionota, Bacteroidota, and Verrucomicrobiota. The population of <em>Vibrio</em> genera were increased significantly, while several other genera, including <em>Denitromonas</em>, <em>Nitrosomonas</em>, <em>Blastopirellula</em>, <em>Fusibacter</em>, <em>Alteromonas</em>, <em>KI89A_clade</em>, and <em>Ruegeria</em>, were decreased significantly after infection. These findings suggest that VP<sub>AHPND</sub> infection has a significant impact on the immune defense and the composition of the copepod microbiota.</div></div>","PeriodicalId":11228,"journal":{"name":"Developmental and comparative immunology","volume":null,"pages":null},"PeriodicalIF":2.7,"publicationDate":"2024-09-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142343554","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-09-27DOI: 10.1016/j.dci.2024.105272
Hui Xu , Wenjuan Dai , Zhengyu Xiong , NaNa Huang , Yanrui Wang , Zhe Yang , Shanshan Luo , Jielian Wu
A cDNA encoding a phage-type lysozyme, designated as HcPLYZ, was successfully cloned from Hyriopsis cumingii. The full-length cDNA sequence of HcPLYZ was determined to be 896 base pairs in length. Analysis revealed the absence of a signal peptide at its N-terminus, and identified two highly conserved phage-type lysozyme activity sites, Glu20 and Asp29, within the deduced amino acid sequence of HcPLYZ. The results of the cloning and sequencing symbiotic bacteria in tissues were consistent with those obtained using tissue cDNA as the template, suggesting that HcPLYZ may originate a symbiotic bacterium. The expression levels of HcPLYZ mRNA exhibited significant variations across different tissues. Successful expression was induced using IPTG, and the native recombinant protein was subsequently purified through affinity chromatography employing Ni2+, and the optimal pH and temperature of which were determined to be 5.5 and 50 °C, respectively. Following exposure to Aeromonas hydrophila, there was a significant increase in the levels of HcPLYZ mRNA in the hemocytes, hepatopancreas, and gills. HcPLYZ was demonstrated the inhibition activity of 55% and 83% against Micrococcus lysodeikticus under pH 5.5 and 50 °C conditions, respectively. These results suggested that HcPLYZ possessed antibacterial activity against both A. hydrophila and M. lysodeikticus.
{"title":"Identification and antibacterial activity of a novel phage-type lysozyme from the freshwater mussel Hyriopsis cumingii","authors":"Hui Xu , Wenjuan Dai , Zhengyu Xiong , NaNa Huang , Yanrui Wang , Zhe Yang , Shanshan Luo , Jielian Wu","doi":"10.1016/j.dci.2024.105272","DOIUrl":"10.1016/j.dci.2024.105272","url":null,"abstract":"<div><div>A cDNA encoding a phage-type lysozyme, designated as HcPLYZ, was successfully cloned from <em>Hyriopsis cumingii</em>. The full-length cDNA sequence of HcPLYZ was determined to be 896 base pairs in length. Analysis revealed the absence of a signal peptide at its N-terminus, and identified two highly conserved phage-type lysozyme activity sites, Glu<sup>20</sup> and Asp<sup>29</sup>, within the deduced amino acid sequence of HcPLYZ. The results of the cloning and sequencing symbiotic bacteria in tissues were consistent with those obtained using tissue cDNA as the template, suggesting that HcPLYZ may originate a symbiotic bacterium. The expression levels of HcPLYZ mRNA exhibited significant variations across different tissues. Successful expression was induced using IPTG, and the native recombinant protein was subsequently purified through affinity chromatography employing Ni<sup>2+</sup>, and the optimal pH and temperature of which were determined to be 5.5 and 50 °C, respectively. Following exposure to <em>Aeromonas hydrophila</em>, there was a significant increase in the levels of HcPLYZ mRNA in the hemocytes, hepatopancreas, and gills. HcPLYZ was demonstrated the inhibition activity of 55% and 83% against <em>Micrococcus lysodeikticus</em> under pH 5.5 and 50 °C conditions, respectively. These results suggested that HcPLYZ possessed antibacterial activity against both <em>A. hydrophila</em> and <em>M. lysodeikticus</em>.</div></div>","PeriodicalId":11228,"journal":{"name":"Developmental and comparative immunology","volume":null,"pages":null},"PeriodicalIF":2.7,"publicationDate":"2024-09-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142343552","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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