Pub Date : 2024-09-28DOI: 10.1016/j.fsi.2024.109927
Penghui Yu , Lei Su , Bo Li , Jianguo Su , Gailing Yuan
Aeromonas hydrophila infection is a severe, acute, and life-threatening disease affecting grass carp (Ctenopharyngodon idella) in aquaculture. Ferroptosis is a novel form of cell death characterized by the accumulation of free iron and harmful lipid peroxides within cells. While selenomethionine (Se-Met) is known to effectively inhibit ferroptosis and alleviate cell damage, its ability to counteract oxidative stress and ferroptosis induced by A. hydrophila remains unclear. The objective of this study is to reveal the possible mechanism behind the ferroptosis phenomenon during A. hydrophila infection. We established a macrophage model of A. hydrophila invasion to monitor the dynamic changes in iron metabolism markers to evaluate the correlation between ferroptotic stress and A. hydrophila infection. A. hydrophila infection induces cytotoxicity and mitochondrial membrane damage via ferroptosis. This damage is attributed to the accumulation of lipid peroxides due to intracellular ferrous ion overload and glutathione depletion. Supplementation of Se-Met reduced mitochondrial damage, enhanced antioxidant enzyme activity and reduced ferroptosis by activating the Nrf2/HO1/GPX4 axis. These findings provide new insights into the regulatory mechanisms of A. hydrophila-induced ferroptosis in teleosts and suggest that targeted inhibition of ferroptosis may offer a novel therapeutic strategy for managing A. hydrophila infections.
{"title":"Selenomethionine alleviates Aeromonas hydrophila-induced oxidative stress and ferroptosis via the Nrf2/HO1/GPX4 pathway in grass carp","authors":"Penghui Yu , Lei Su , Bo Li , Jianguo Su , Gailing Yuan","doi":"10.1016/j.fsi.2024.109927","DOIUrl":"10.1016/j.fsi.2024.109927","url":null,"abstract":"<div><div><em>Aeromonas hydrophila</em> infection is a severe, acute, and life-threatening disease affecting grass carp (<em>Ctenopharyngodon idella</em>) in aquaculture. Ferroptosis is a novel form of cell death characterized by the accumulation of free iron and harmful lipid peroxides within cells. While selenomethionine (Se-Met) is known to effectively inhibit ferroptosis and alleviate cell damage, its ability to counteract oxidative stress and ferroptosis induced by <em>A. hydrophila</em> remains unclear. The objective of this study is to reveal the possible mechanism behind the ferroptosis phenomenon during <em>A. hydrophila</em> infection. We established a macrophage model of <em>A. hydrophila</em> invasion to monitor the dynamic changes in iron metabolism markers to evaluate the correlation between ferroptotic stress and <em>A. hydrophila</em> infection. <em>A. hydrophila</em> infection induces cytotoxicity and mitochondrial membrane damage via ferroptosis. This damage is attributed to the accumulation of lipid peroxides due to intracellular ferrous ion overload and glutathione depletion. Supplementation of Se-Met reduced mitochondrial damage, enhanced antioxidant enzyme activity and reduced ferroptosis by activating the Nrf2/HO1/GPX4 axis. These findings provide new insights into the regulatory mechanisms of <em>A. hydrophila</em>-induced ferroptosis in teleosts and suggest that targeted inhibition of ferroptosis may offer a novel therapeutic strategy for managing <em>A. hydrophila</em> infections.</div></div>","PeriodicalId":12127,"journal":{"name":"Fish & shellfish immunology","volume":"154 ","pages":"Article 109927"},"PeriodicalIF":4.1,"publicationDate":"2024-09-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142344318","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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 : 2024-09-28DOI: 10.1016/j.fsi.2024.109933
Decheng Wang , Tianqi Wang , Daniel Kim , Shelly Tan , Sheng Liu , Jun Wan , Qing Deng
Neutrophils are crucial for defense against numerous infections, and their migration and activations are tightly regulated to prevent collateral tissue damage. We previously performed a neutrophil-specific miRNA overexpression screening and identified several microRNAs, including miR-375, as potent modulators for neutrophil activity. Overexpression of miR-375 decreases neutrophil motility and migration in zebrafish and human neutrophil-like cells. We screened the genes downregulated by miR-375 in zebrafish neutrophils and identified that Cathepsin B (Ctsba) is required for neutrophil motility and chemotaxis upon tail wounding and bacterial infection. Pharmacological inhibition or neutrophil-specific knockout of ctsba significantly decreased the neutrophil chemotaxis in zebrafish and survival upon systemic bacterial infection. Notably, Ctsba knockdown in human neutrophil-like cells also resulted in reduced chemotaxis. Inhibiting integrin receptor function using RGDS rescued the neutrophil migration defects and susceptibility to systemic infection in zebrafish with either miR-375 overexpression or ctsba knockout. Our results demonstrate that miR-375 and its target Ctsba modulate neutrophil activity during tissue injury and bacterial infection in vivo, providing novel insights into neutrophil biology and the overall inflammation process.
{"title":"MicroRNA-375 modulates neutrophil chemotaxis via targeting Cathepsin B in zebrafish","authors":"Decheng Wang , Tianqi Wang , Daniel Kim , Shelly Tan , Sheng Liu , Jun Wan , Qing Deng","doi":"10.1016/j.fsi.2024.109933","DOIUrl":"10.1016/j.fsi.2024.109933","url":null,"abstract":"<div><div>Neutrophils are crucial for defense against numerous infections, and their migration and activations are tightly regulated to prevent collateral tissue damage. We previously performed a neutrophil-specific miRNA overexpression screening and identified several microRNAs, including miR-375, as potent modulators for neutrophil activity. Overexpression of miR-375 decreases neutrophil motility and migration in zebrafish and human neutrophil-like cells. We screened the genes downregulated by miR-375 in zebrafish neutrophils and identified that Cathepsin B (Ctsba) is required for neutrophil motility and chemotaxis upon tail wounding and bacterial infection. Pharmacological inhibition or neutrophil-specific knockout of <em>ctsba</em> significantly decreased the neutrophil chemotaxis in zebrafish and survival upon systemic bacterial infection. Notably, Ctsba knockdown in human neutrophil-like cells also resulted in reduced chemotaxis. Inhibiting integrin receptor function using RGDS rescued the neutrophil migration defects and susceptibility to systemic infection in zebrafish with either miR-375 overexpression or <em>ctsba</em> knockout. Our results demonstrate that miR-375 and its target Ctsba modulate neutrophil activity during tissue injury and bacterial infection <em>in vivo</em>, providing novel insights into neutrophil biology and the overall inflammation process.</div></div>","PeriodicalId":12127,"journal":{"name":"Fish & shellfish immunology","volume":"154 ","pages":"Article 109933"},"PeriodicalIF":4.1,"publicationDate":"2024-09-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142344315","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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.fsi.2024.109932
Alexandra Y. Andreyeva , Tatyana A. Kukhareva , Olga L. Gostyukhina , Oksana Y. Vialova , Anastasia A. Tkachuk , Elina S. Chelebieva , Maria S. Podolskaya , Andrey B. Borovkov , Elizaveta A. Bogacheva , Daria S. Lavrichenko , Ekaterina S. Kladchenko
There is growing recognition that the hypoxic regions of the ocean are also becoming more acidic due to increasing levels of global carbon dioxide emissions. The impact of water acidification on marine life is largely unknown, as most previous studies have not taken into account the effects of hypoxia, which may affect how organisms respond to low pH levels. In this study, we experimentally examined the consequences of water acidification in combination with normoxic or hypoxic conditions on cellular immune parameters in Mediterranean mussels. We measured total hemocyte counts in hemolymph, the cellular composition of hemolymph, phagocytosis, reactive oxygen species (ROS) production. General response of the organism was evaluated on the basis of the activity of antioxidant enzymes in the hepatopancreas, as well as respiratory rates over an 8-day exposure period. The mussels were exposed to low pH conditions (7.3), either under normoxic conditions (dissolved oxygen concentration of 8 mg/L) or hypoxic conditions (dissolved oxygen concentration of 2 mg/L). The parameters were assessed at days 1, 3, 6, and 8 of the experiment. Experimental acidification under normoxic conditions reduced THC and ROS production by hemocytes during later stages of exposure, but phagocytic activity (PA) only decreased at day 3 and then recovered. Combined acidification and hypoxia suppressed PA in hemocytes at the beginning of exposure, while hemocyte ROS production and THC decreased by the end of the experiment. The hemolymph cellular composition and activity of antioxidant enzymes were unaffected by acidified conditions under different oxygen regimes, but mussel respiratory rate (RR) decreased with a more significant reduction in oxygen consumption under hypoxia. Mussels showed a relatively high tolerance to acidification in combination with various dissolved oxygen levels, although prolonged acidification exposure led to increased detrimental effects on immunity and metabolism.
{"title":"Impacts of ocean acidification and hypoxia on cellular immunity, oxygen consumption and antioxidant status in Mediterranean mussel","authors":"Alexandra Y. Andreyeva , Tatyana A. Kukhareva , Olga L. Gostyukhina , Oksana Y. Vialova , Anastasia A. Tkachuk , Elina S. Chelebieva , Maria S. Podolskaya , Andrey B. Borovkov , Elizaveta A. Bogacheva , Daria S. Lavrichenko , Ekaterina S. Kladchenko","doi":"10.1016/j.fsi.2024.109932","DOIUrl":"10.1016/j.fsi.2024.109932","url":null,"abstract":"<div><div>There is growing recognition that the hypoxic regions of the ocean are also becoming more acidic due to increasing levels of global carbon dioxide emissions. The impact of water acidification on marine life is largely unknown, as most previous studies have not taken into account the effects of hypoxia, which may affect how organisms respond to low pH levels. In this study, we experimentally examined the consequences of water acidification in combination with normoxic or hypoxic conditions on cellular immune parameters in Mediterranean mussels. We measured total hemocyte counts in hemolymph, the cellular composition of hemolymph, phagocytosis, reactive oxygen species (ROS) production. General response of the organism was evaluated on the basis of the activity of antioxidant enzymes in the hepatopancreas, as well as respiratory rates over an 8-day exposure period. The mussels were exposed to low pH conditions (7.3), either under normoxic conditions (dissolved oxygen concentration of 8 mg/L) or hypoxic conditions (dissolved oxygen concentration of 2 mg/L). The parameters were assessed at days 1, 3, 6, and 8 of the experiment. Experimental acidification under normoxic conditions reduced THC and ROS production by hemocytes during later stages of exposure, but phagocytic activity (PA) only decreased at day 3 and then recovered. Combined acidification and hypoxia suppressed PA in hemocytes at the beginning of exposure, while hemocyte ROS production and THC decreased by the end of the experiment. The hemolymph cellular composition and activity of antioxidant enzymes were unaffected by acidified conditions under different oxygen regimes, but mussel respiratory rate (RR) decreased with a more significant reduction in oxygen consumption under hypoxia. Mussels showed a relatively high tolerance to acidification in combination with various dissolved oxygen levels, although prolonged acidification exposure led to increased detrimental effects on immunity and metabolism.</div></div>","PeriodicalId":12127,"journal":{"name":"Fish & shellfish immunology","volume":"154 ","pages":"Article 109932"},"PeriodicalIF":4.1,"publicationDate":"2024-09-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142344313","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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.fsi.2024.109931
Feng-yuan Yan , Yuan-feng Xu , Wen-rong Feng , Qing-hong He , Guo-an Hua , Wen-jing Li , Pao Xu , Jun Zhou , Yong-kai Tang
Hypoxic stress, triggered by a multitude of factors, has inflicted significant economic repercussions on the aquaculture of Eriocheir sinensis. In this research, we sequenced a collective of 60 samples from both hypoxia-sensitive and hypoxia-resistant groups utilizing streamlined genome sequencing techniques. Subsequently, we delved into population evolution, scrutinized the selective sweep within these populations, and performed a genome-wide association study (GWAS) focused on the hypoxia tolerance traits within the population, all through the lens of SNPs molecular markers. This comprehensive analysis aimed to uncover the SNPs and pinpoint the pertinent candidate genes that influence the hypoxia tolerance capabilities of E. sinensis. The selective sweep analysis revealed that genes harboring potential genetic variations within the two populations were predominantly enriched in areas such as signaling molecules and interactions, energy metabolism, glycolipid metabolism, and immune response. In the genome-wide association study focusing on hypoxia tolerance traits, we identified four SNPs significantly associated with hypoxia resistance. Furthermore, one potential candidate gene, Dscam2, which is believed to influence hypoxia tolerance, was discovered within a 50 kb vicinity of these SNPs. These identified SNPs can serve as molecular markers for screening hypoxia tolerance, offering valuable insights for the genetic improvement of E. sinensis.
{"title":"Genomic analysis of hypoxia-tolerant population of the Chinese mitten crab (Eriocheir sinensis)","authors":"Feng-yuan Yan , Yuan-feng Xu , Wen-rong Feng , Qing-hong He , Guo-an Hua , Wen-jing Li , Pao Xu , Jun Zhou , Yong-kai Tang","doi":"10.1016/j.fsi.2024.109931","DOIUrl":"10.1016/j.fsi.2024.109931","url":null,"abstract":"<div><div>Hypoxic stress, triggered by a multitude of factors, has inflicted significant economic repercussions on the aquaculture of <em>Eriocheir sinensis</em>. In this research, we sequenced a collective of 60 samples from both hypoxia-sensitive and hypoxia-resistant groups utilizing streamlined genome sequencing techniques. Subsequently, we delved into population evolution, scrutinized the selective sweep within these populations, and performed a genome-wide association study (GWAS) focused on the hypoxia tolerance traits within the population, all through the lens of SNPs molecular markers. This comprehensive analysis aimed to uncover the SNPs and pinpoint the pertinent candidate genes that influence the hypoxia tolerance capabilities of <em>E. sinensis</em>. The selective sweep analysis revealed that genes harboring potential genetic variations within the two populations were predominantly enriched in areas such as signaling molecules and interactions, energy metabolism, glycolipid metabolism, and immune response. In the genome-wide association study focusing on hypoxia tolerance traits, we identified four SNPs significantly associated with hypoxia resistance. Furthermore, one potential candidate gene, <em>Dscam2</em>, which is believed to influence hypoxia tolerance, was discovered within a 50 kb vicinity of these SNPs. These identified SNPs can serve as molecular markers for screening hypoxia tolerance, offering valuable insights for the genetic improvement of <em>E. sinensis</em>.</div></div>","PeriodicalId":12127,"journal":{"name":"Fish & shellfish immunology","volume":"154 ","pages":"Article 109931"},"PeriodicalIF":4.1,"publicationDate":"2024-09-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142344312","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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 : 2024-09-26DOI: 10.1016/j.fsi.2024.109930
Da Huang , Rui Cheng , Xiaodan Liu , JiaQi Zhang , Chi Zhang
Leader RNAs are viral small non-coding RNAs that has been proved to play important roles in viral replication. Snakehead vesiculovirus (SHVV) is an aquatic virus that has caused huge economic loss in Chinese snakehead fish aquaculture industry. It has been proved that SHVV would generate leader RNA during the process of infection, and leader RNA could interact with viral nucleoprotein to promote viral replication. In this study, we identified that leader RNA could also interact with cellular protein Cold Shock Domain containing E1 (CSDE1) and heterogeneous nuclear ribonucleoproteins A3 (hnRNP A3). Further investigation reveals that overexpression of CSDE1 and hnRNP A3 facilitated SHVV replication. Downregulation of CSDE1 and hnRNP A3 by siRNA inhibited SHVV replication. This study provided a new sight into understand the mechanism of SHVV replication, and a potential anti-SHVV target for drug research.
{"title":"Leader RNA facilitates snakehead vesiculovirus (SHVV) replication by interacting with CSDE1 and hnRNP A3","authors":"Da Huang , Rui Cheng , Xiaodan Liu , JiaQi Zhang , Chi Zhang","doi":"10.1016/j.fsi.2024.109930","DOIUrl":"10.1016/j.fsi.2024.109930","url":null,"abstract":"<div><div>Leader RNAs are viral small non-coding RNAs that has been proved to play important roles in viral replication. Snakehead vesiculovirus (SHVV) is an aquatic virus that has caused huge economic loss in Chinese snakehead fish aquaculture industry. It has been proved that SHVV would generate leader RNA during the process of infection, and leader RNA could interact with viral nucleoprotein to promote viral replication. In this study, we identified that leader RNA could also interact with cellular protein Cold Shock Domain containing E1 (CSDE1) and heterogeneous nuclear ribonucleoproteins A3 (hnRNP A3). Further investigation reveals that overexpression of CSDE1 and hnRNP A3 facilitated SHVV replication. Downregulation of CSDE1 and hnRNP A3 by siRNA inhibited SHVV replication. This study provided a new sight into understand the mechanism of SHVV replication, and a potential anti-SHVV target for drug research.</div></div>","PeriodicalId":12127,"journal":{"name":"Fish & shellfish immunology","volume":"154 ","pages":"Article 109930"},"PeriodicalIF":4.1,"publicationDate":"2024-09-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142344314","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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 : 2024-09-26DOI: 10.1016/j.fsi.2024.109929
Hongzhou Xu , Jingfei Long , Xiaoyu Qi , Ping Li , Chenyang Yan , Lixin Wang , Yuanjiang Jin , Haixia Liu
Galectin-9 (Gal-9) belongs to a family of the glycan-binding proteins (GBPs) and is known to restrict bacterial activity via interacting with pathogen associated molecular pattern (PAMPs). However, the underlying immune mechanism of endogenous Gal-9 on fish against bacterial infection is still unclear. In this study, effects of Gal-9 from Onychostoma macrolepis (OmGal-9) on expression of immune-related genes were measured by HEK293T. The immune response of O. macrolepis with OmGal-9 overexpression to Aeromonas hydrophila (A. hydrophila) infection (1.65 × 108 CFU/mL) was evaluated by tissue bacterial load, fish survival rate and transcriptome analysis. The results showed that OmGal-9 displayed a punctate distribution in the nucleus and cytoplasm of HEK293T cells. Compared to cells transfected with the empty vector (EV group), recombinant plasmid pEGFP-Gal9 treatment (Gal9 group) significantly down-regulated the expression of immune-related genes TNFα, STAT3, MyD88, LCK, and p52 of HEK293T cells stimulated with LPS at 24 h, while up-regulated IκBα and caspase-1 (P < 0.05). The activities of catalase (CAT), superoxide dismutase (SOD), the total antioxidant capacity (T-AOC), alkaline phosphatase (AKP), acid phosphatase (ACP), and lysozyme (LZM) of O. macrolepis were significantly increased on 7 days in Gal9 group compared to EV group (P < 0.05). The bacterial load of liver, spleen, and kidney of O. macrolepis infected with A. hydrophila in Gal9 group at 24 h was significantly lower than that in EV group (P < 0.05), and the survival rate had increased from 15 % to 35 %. A comparative transcriptome analysis between the Gal9 and EV group identified 305 differentially expressed genes (DEGs). The analysis showed that OmGal-9 might play an important regulatory role in glycolysis/gluconeogenesis, fatty acid degradation, and ascorbate and aldarate metabolism. Moreover, the immune-related DEGs were predominantly enriched in eleven pathways, with the most important three of them being linked to innate immunity: NOD-like, C-type lectin and Toll-like receptor signaling pathway. Taking together, OmGal-9 can enhance the resistance of fish to bacterial diseases by improving immune system function and activating immune-related pathways.
{"title":"Galectin-9 activates host immune response and improve immunoprotection of Onychostoma macrolepis against Aeromonas hydrophila infection","authors":"Hongzhou Xu , Jingfei Long , Xiaoyu Qi , Ping Li , Chenyang Yan , Lixin Wang , Yuanjiang Jin , Haixia Liu","doi":"10.1016/j.fsi.2024.109929","DOIUrl":"10.1016/j.fsi.2024.109929","url":null,"abstract":"<div><div>Galectin-9 (Gal-9) belongs to a family of the glycan-binding proteins (GBPs) and is known to restrict bacterial activity via interacting with pathogen associated molecular pattern (PAMPs). However, the underlying immune mechanism of endogenous Gal-9 on fish against bacterial infection is still unclear. In this study, effects of Gal-9 from <em>Onychostoma macrolepis</em> (<em>Om</em>Gal-9) on expression of immune-related genes were measured by HEK293T. The immune response of <em>O. macrolepis</em> with <em>Om</em>Gal-9 overexpression to <em>Aeromonas hydrophila</em> (<em>A. hydrophila</em>) infection (1.65 × 10<sup>8</sup> CFU/mL) was evaluated by tissue bacterial load, fish survival rate and transcriptome analysis. The results showed that <em>Om</em>Gal-9 displayed a punctate distribution in the nucleus and cytoplasm of HEK293T cells. Compared to cells transfected with the empty vector (EV group), recombinant plasmid pEGFP-Gal9 treatment (Gal9 group) significantly down-regulated the expression of immune-related genes <em>TNFα</em>, <em>STAT3</em>, <em>MyD88, LCK,</em> and <em>p52</em> of HEK293T cells stimulated with LPS at 24 h, while up-regulated <em>IκBα</em> and <em>caspase</em><em>-</em><em>1</em> (<em>P</em> < 0.05). The activities of catalase (CAT), superoxide dismutase (SOD), the total antioxidant capacity (T-AOC), alkaline phosphatase (AKP), acid phosphatase (ACP), and lysozyme (LZM) of <em>O. macrolepis</em> were significantly increased on 7 days in Gal9 group compared to EV group (<em>P</em> < 0.05). The bacterial load of liver, spleen, and kidney of <em>O. macrolepis</em> infected with <em>A. hydrophila</em> in Gal9 group at 24 h was significantly lower than that in EV group (<em>P</em> < 0.05), and the survival rate had increased from 15 % to 35 %. A comparative transcriptome analysis between the Gal9 and EV group identified 305 differentially expressed genes (DEGs). The analysis showed that <em>Om</em>Gal-9 might play an important regulatory role in glycolysis/gluconeogenesis, fatty acid degradation, and ascorbate and aldarate metabolism. Moreover, the immune-related DEGs were predominantly enriched in eleven pathways, with the most important three of them being linked to innate immunity: NOD-like, C-type lectin and Toll-like receptor signaling pathway. Taking together, <em>Om</em>Gal-9 can enhance the resistance of fish to bacterial diseases by improving immune system function and activating immune-related pathways.</div></div>","PeriodicalId":12127,"journal":{"name":"Fish & shellfish immunology","volume":"154 ","pages":"Article 109929"},"PeriodicalIF":4.1,"publicationDate":"2024-09-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142344311","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
The microsporidian Enterocytozoon hepatopenaei (EHP) is an emerging pathogen that causes high economic losses in shrimp industry. The knowledge on shrimp's immune response to EHP infection to properly handle this outbreak is poorly understood. The prophenoloxidase (proPO)-activating system is an important invertebrate innate immune systems that produces melanin and toxic reactive intermediates against invading pathogens. In this study, we investigated the role of the proPO-activating system during EHP infection in the Pacific white shrimp Litopenaeus vannamei. The expression of proPO-activating system-related genes was highly responded to the EHP infection and the hemolymph PO activity was significantly increased and tightly regulated during the infection. The melanization products, generated by the proPO activation, exhibit in vitro cytotoxicity effect on the EHP spores and inhibit spore germination. Suppression of the proPO-activating system by RNA interference significantly decreased hemolymph PO activity and resulted in increased EHP copy number and reduced expression of several genes in the JAK/STAT and Toll signaling pathways as well as antimicrobial peptides. Furthermore, suppression of the proPO system also reduced hemocyte adhesion and encapsulation of the EHP spores. These results demonstrated that the proPO system plays a vital role in reducing EHP infectivity and cross-talks with other humoral and cellular responses to coordinately defend the EHP infection.
{"title":"Prophenoloxidase-activating system plays a crucial role in innate immune responses to Enterocytozoon hepatopenaei infection in shrimp Litopenaeus vannamei","authors":"Pongsakorn Sukonthamarn, Pavarisa Wongvises, Nutthapon Sangklai, Pattana Jaroenlak, Anchalee Tassanakajon","doi":"10.1016/j.fsi.2024.109925","DOIUrl":"10.1016/j.fsi.2024.109925","url":null,"abstract":"<div><div>The microsporidian <em>Enterocytozoon hepatopenaei</em> (EHP) is an emerging pathogen that causes high economic losses in shrimp industry. The knowledge on shrimp's immune response to EHP infection to properly handle this outbreak is poorly understood. The prophenoloxidase (proPO)-activating system is an important invertebrate innate immune systems that produces melanin and toxic reactive intermediates against invading pathogens. In this study, we investigated the role of the proPO-activating system during EHP infection in the Pacific white shrimp <em>Litopenaeus vannamei</em>. The expression of proPO-activating system-related genes was highly responded to the EHP infection and the hemolymph PO activity was significantly increased and tightly regulated during the infection. The melanization products, generated by the proPO activation, exhibit <em>in vitro</em> cytotoxicity effect on the EHP spores and inhibit spore germination. Suppression of the proPO-activating system by RNA interference significantly decreased hemolymph PO activity and resulted in increased EHP copy number and reduced expression of several genes in the JAK/STAT and Toll signaling pathways as well as antimicrobial peptides. Furthermore, suppression of the proPO system also reduced hemocyte adhesion and encapsulation of the EHP spores. These results demonstrated that the proPO system plays a vital role in reducing EHP infectivity and cross-talks with other humoral and cellular responses to coordinately defend the EHP infection.</div></div>","PeriodicalId":12127,"journal":{"name":"Fish & shellfish immunology","volume":"154 ","pages":"Article 109925"},"PeriodicalIF":4.1,"publicationDate":"2024-09-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142344317","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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 : 2024-09-25DOI: 10.1016/j.fsi.2024.109924
Shouhu Li , Chuang Qin , Shi-Ming Peng , Ya-Bing Wang , Yuan Wang , Xiao-Shan Wang , Jian-Gao Shi , Xin-Cang Li
The phospholipase A2 (PLA2) superfamily has attracted increasing attention in recent years due to the multiple physiological and pathological functions exerted by its members. Up to date, the knowledge about the biological role of PLA2XIIA subfamily members remains limited. In this study, a new member of PLA2XIIA subfamily, LcPLA2XIIA, was characterized in large yellow croaker. Different from most members of the PLA2 superfamily with positive charge, LcPLA2XIIA encodes an anionic protein, which is similar to other members of PLA2XIIA subfamily. LcPLA2XIIA is highly expressed in the intestine, and afterwards, it is up-regulated after with Pseudomonas plecoglossicida or Staphylococcus aureus. LcPLA2XIIA exhibits strong inhibitory activity against these two bacteria. The results indicate that LcPLA2XIIA plays an important role in the antimicrobial immune responses of large yellow croaker. LcPLA2XIIA displays strong binding activity to all the tested bacteria. It specifically interacts with LTA, a unique component on the surface of Gram-positive bacteria. It also significantly promotes bacterial agglutination in the presence of Ca2+. These findings reveal that the binding and agglutinating abilities of LcPLA2XIIA to bacteria contribute greatly to its antibacterial activity. In addition, LcPLA2XIIA significantly inhibits the proliferation of infectious hematopoietic necrosis virus instead of recombinant human adenovirus type 5. It also suppresses the growth of human colorectal adenocarcinoma cells by inducing apoptosis, but it has no obvious inhibitory effect on the growth of epithelioma papulosum cyprinid cells. This study provides new insights into the antibacterial activity, and the mechanism of LcPLA2XIIA in large yellow croaker, and antiviral and antitumor functions of PLA2XIIA subfamily members.
{"title":"A newly identified secretory phospholipase A2 group XIIA homolog (LcPLA2XIIA) in Larimichthys crocea exhibits antimicrobial and antitumor activities","authors":"Shouhu Li , Chuang Qin , Shi-Ming Peng , Ya-Bing Wang , Yuan Wang , Xiao-Shan Wang , Jian-Gao Shi , Xin-Cang Li","doi":"10.1016/j.fsi.2024.109924","DOIUrl":"10.1016/j.fsi.2024.109924","url":null,"abstract":"<div><div>The phospholipase A2 (PLA2) superfamily has attracted increasing attention in recent years due to the multiple physiological and pathological functions exerted by its members. Up to date, the knowledge about the biological role of PLA2XIIA subfamily members remains limited. In this study, a new member of PLA2XIIA subfamily, <em>Lc</em>PLA2XIIA, was characterized in large yellow croaker. Different from most members of the PLA2 superfamily with positive charge, <em>LcPLA2XIIA</em> encodes an anionic protein, which is similar to other members of PLA2XIIA subfamily. <em>LcPLA2XIIA</em> is highly expressed in the intestine, and afterwards, it is up-regulated after with <em>Pseudomonas plecoglossicida</em> or <em>Staphylococcus aureus</em>. <em>Lc</em>PLA2XIIA exhibits strong inhibitory activity against these two bacteria. The results indicate that <em>Lc</em>PLA2XIIA plays an important role in the antimicrobial immune responses of large yellow croaker. <em>Lc</em>PLA2XIIA displays strong binding activity to all the tested bacteria. It specifically interacts with LTA, a unique component on the surface of Gram-positive bacteria. It also significantly promotes bacterial agglutination in the presence of Ca<sup>2+</sup>. These findings reveal that the binding and agglutinating abilities of <em>Lc</em>PLA2XIIA to bacteria contribute greatly to its antibacterial activity. In addition, <em>Lc</em>PLA2XIIA significantly inhibits the proliferation of infectious hematopoietic necrosis virus instead of recombinant human adenovirus type 5. It also suppresses the growth of human colorectal adenocarcinoma cells by inducing apoptosis, but it has no obvious inhibitory effect on the growth of epithelioma papulosum cyprinid cells. This study provides new insights into the antibacterial activity, and the mechanism of <em>Lc</em>PLA2XIIA in large yellow croaker, and antiviral and antitumor functions of PLA2XIIA subfamily members.</div></div>","PeriodicalId":12127,"journal":{"name":"Fish & shellfish immunology","volume":"154 ","pages":"Article 109924"},"PeriodicalIF":4.1,"publicationDate":"2024-09-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142344310","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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 : 2024-09-25DOI: 10.1016/j.fsi.2024.109928
Huan Wang , Xu Zhang , Zixuan Wang , Lipeng Shan , Shiyi Zhu , Guanglu Liu , Lei Liu , Yang Hu , Jiong Chen
Micropterus salmoides rhabdovirus (MSRV) poses a significant threat to aquaculture, causing substantial economic losses. In this study, we evaluated the antiviral efficacy and immunomodulatory potential of palmatine, a plant-derived monomer, against MSRV infection in largemouth bass. Our results demonstrated that palmatine significantly inhibited MSRV replication, with a reduction in viral nucleoprotein expression by 85 % at a safe concentration. Additionally, palmatine pre-treatment of EPC cells enhanced their antiviral capacity, with a maximum inhibition rate of 82 % following 24 h pre-incubation. Palmatine also effectively reduced MSRV-induced cytopathic effects, protecting cellular integrity and maintaining mitochondrial membrane potential. In vivo studies revealed that palmatine immersion at 80 mg/L was non-toxic and significantly suppressed MSRV replication in largemouth bass, increasing survival rates by 53 % over 15 d. Furthermore, palmatine pre-treatment enhanced the fish's resistance to MSRV, with a 78 % inhibition rate of viral replication and a 46 % increase in survival rate. Mechanistically, palmatine activated key immune genes, including IRF3, IRF7, and IFN, indicating its role in boosting innate immune responses. The compound also reduced horizontal transmission of MSRV in a cohabitation model, decreasing viral spread by up to 78 % over nine days. These findings highlight palmatine's potential as a small-molecule immunomodulator in aquaculture, offering a sustainable approach to disease management and enhancing fish health and welfare. Integrating palmatine into fish diets as an immunostimulant could provide a continuous, proactive defense against viral outbreaks, promoting more resilient and sustainable aquaculture practices.
{"title":"Palmatine as a potent immunomodulator: Enhancing resistance to Micropterus salmoides rhabdovirus in largemouth bass through innate immune activation and viral suppression","authors":"Huan Wang , Xu Zhang , Zixuan Wang , Lipeng Shan , Shiyi Zhu , Guanglu Liu , Lei Liu , Yang Hu , Jiong Chen","doi":"10.1016/j.fsi.2024.109928","DOIUrl":"10.1016/j.fsi.2024.109928","url":null,"abstract":"<div><div><em>Micropterus salmoides</em> rhabdovirus (MSRV) poses a significant threat to aquaculture, causing substantial economic losses. In this study, we evaluated the antiviral efficacy and immunomodulatory potential of palmatine, a plant-derived monomer, against MSRV infection in largemouth bass. Our results demonstrated that palmatine significantly inhibited MSRV replication, with a reduction in viral nucleoprotein expression by 85 % at a safe concentration. Additionally, palmatine pre-treatment of EPC cells enhanced their antiviral capacity, with a maximum inhibition rate of 82 % following 24 h pre-incubation. Palmatine also effectively reduced MSRV-induced cytopathic effects, protecting cellular integrity and maintaining mitochondrial membrane potential. <em>In vivo</em> studies revealed that palmatine immersion at 80 mg/L was non-toxic and significantly suppressed MSRV replication in largemouth bass, increasing survival rates by 53 % over 15 d. Furthermore, palmatine pre-treatment enhanced the fish's resistance to MSRV, with a 78 % inhibition rate of viral replication and a 46 % increase in survival rate. Mechanistically, palmatine activated key immune genes, including <em>IRF3</em>, <em>IRF7</em>, and <em>IFN</em>, indicating its role in boosting innate immune responses. The compound also reduced horizontal transmission of MSRV in a cohabitation model, decreasing viral spread by up to 78 % over nine days. These findings highlight palmatine's potential as a small-molecule immunomodulator in aquaculture, offering a sustainable approach to disease management and enhancing fish health and welfare. Integrating palmatine into fish diets as an immunostimulant could provide a continuous, proactive defense against viral outbreaks, promoting more resilient and sustainable aquaculture practices.</div></div>","PeriodicalId":12127,"journal":{"name":"Fish & shellfish immunology","volume":"154 ","pages":"Article 109928"},"PeriodicalIF":4.1,"publicationDate":"2024-09-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142327025","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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 : 2024-09-24DOI: 10.1016/j.fsi.2024.109922
Xianhui Wang , Pengfei Mu , Jieying Huo , Feng Han , Xinhua Chen , Jingqun Ao
Peroxiredoxin IV (PrxIV), which possesses an N-terminal signal peptide, is the only secretable protein in Prx family. PrxIV can protect cells against reactive oxygen species (ROS) and act as a DAMP to promote infection-independent immune response. However, the characterization and regulation of promoters of PrxIV genes are rarely reported. In this study, a 1511-bp 5′-flanking sequence of large yellow croaker (Larimichthys crocea) PrxIV (LcPrxIV) was cloned and characterized. DNA truncation combined with luciferase activity assay revealed that a fragment of −781/+20 contained in the plasmid LcPrxIV-P3 exhibited the highest promoter activity. It could initiate the luciferase expression up to 44.6-fold when compared to control plasmid pGL3-Basic. TFSEARCH analysis revealed many recognizing sequences of transcriptional factors exist within this 1511-bp sequence, including Foxo and CREB. Altogether, four putative binding sites located in three recognizing sequences of CREB were identified. Notably, co-transfection of LcPrxIV-P3 with LcCREB led to a significant 2.48-fold increase of the LcPrxIV-P3 promoter activity (P<0.01). Furthermore, the mutation at putative binding sites A, B, and all four sites of CREB in the LcPrxIV-P3 caused the significant decrease of activation on LcPrxIV-P3 promoter activity, suggesting these two sites may be the main binding sites of CREB in LcPrxIV promoter. In addition, the oxidative stress caused by hydrogen peroxide, rather than immune stimuli such as Poly (I: C), LPS, LTA, or PGN could lead to the elevation of LcPrxIV-P3 promoter activity. When the concentration of hydrogen peroxide reached 500 μM, the promoter activity of LcPrxIV-P3 could be up-regulated to 1.47-fold, which was extremely significantly different from the control (P<0.001). These results help to elucidate the regulatory mechanisms of LcPrxIV gene expression, and the role of LcPrxIV in protecting cells against oxidative stress or in oxidoreduction-dependent signal transduction.
{"title":"Functional characterization of large yellow croaker (Larimichthys crocea) Peroxiredoxin IV (PrxIV) gene promoter","authors":"Xianhui Wang , Pengfei Mu , Jieying Huo , Feng Han , Xinhua Chen , Jingqun Ao","doi":"10.1016/j.fsi.2024.109922","DOIUrl":"10.1016/j.fsi.2024.109922","url":null,"abstract":"<div><div>Peroxiredoxin IV (PrxIV), which possesses an N-terminal signal peptide, is the only secretable protein in Prx family. PrxIV can protect cells against reactive oxygen species (ROS) and act as a DAMP to promote infection-independent immune response. However, the characterization and regulation of promoters of PrxIV genes are rarely reported. In this study, a 1511-bp 5′-flanking sequence of large yellow croaker (<em>Larimichthys crocea</em>) PrxIV (<em>Lc</em>PrxIV) was cloned and characterized. DNA truncation combined with luciferase activity assay revealed that a fragment of −781/+20 contained in the plasmid <em>Lc</em>PrxIV-P3 exhibited the highest promoter activity. It could initiate the luciferase expression up to 44.6-fold when compared to control plasmid pGL3-Basic. TFSEARCH analysis revealed many recognizing sequences of transcriptional factors exist within this 1511-bp sequence, including Foxo and CREB. Altogether, four putative binding sites located in three recognizing sequences of CREB were identified. Notably, co-transfection of <em>Lc</em>PrxIV-P3 with <em>Lc</em>CREB led to a significant 2.48-fold increase of the <em>Lc</em>PrxIV-P3 promoter activity (<em>P<</em>0.01). Furthermore, the mutation at putative binding sites A, B, and all four sites of CREB in the <em>Lc</em>PrxIV-P3 caused the significant decrease of activation on <em>Lc</em>PrxIV-P3 promoter activity, suggesting these two sites may be the main binding sites of CREB in <em>Lc</em>PrxIV promoter. In addition, the oxidative stress caused by hydrogen peroxide, rather than immune stimuli such as Poly (I: C), LPS, LTA, or PGN could lead to the elevation of <em>Lc</em>PrxIV-P3 promoter activity. When the concentration of hydrogen peroxide reached 500 μM, the promoter activity of <em>Lc</em>PrxIV-P3 could be up-regulated to 1.47-fold, which was extremely significantly different from the control (<em>P<</em>0.001). These results help to elucidate the regulatory mechanisms of <em>Lc</em>PrxIV gene expression, and the role of <em>Lc</em>PrxIV in protecting cells against oxidative stress or in oxidoreduction-dependent signal transduction.</div></div>","PeriodicalId":12127,"journal":{"name":"Fish & shellfish immunology","volume":"154 ","pages":"Article 109922"},"PeriodicalIF":4.1,"publicationDate":"2024-09-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142344319","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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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