Pub Date : 2024-11-01Epub Date: 2024-10-29DOI: 10.1016/j.fsi.2024.109993
Ting Xue, Yanmin Wang, Yiping Liu, Yuping Liu, Chao Li
As a commercially valuable fish species, Sebastes schlegelii faces threats from pathogenic bacteria like Edwardsiella piscicida during aquaculture. The global host range of E. piscicida encompasses various species, yet its pathogenic mechanism remains incompletely elucidated. Cell lines offer invaluable in vitro resources for studying the pathogen pathogenicity. Here, we established and characterized a cell line derived from the intestinal tissue of the S. schlegelii, designated as SSI. SSI has undergone continuous subculturing for over 80 passages, demonstrating robust growth in DMEM supplemented with 10%-20% FBS and 20 μM HEPES at 24°C. Karyotype analysis and 18S rRNA amplification confirm its origin. SSI exhibits high transfection efficiency for exogenous DNA, making it suitable for gene expression and intestinal function analysis. E. piscicida infects SSI cells at low densities without inducing morphological changes within 6 h of infection, suggesting the potential of SSI as an in vitro model for studying E. piscicida pathogenicity. This cell line provides a valuable tool for investigating mucosal immunity and E. piscicida pathogenic mechanisms in marine fish.
{"title":"Establishment and characterization of SSI cell line from Sebastes schlegelii intestine for investigating the immune response to Pathogenic Bacteria.","authors":"Ting Xue, Yanmin Wang, Yiping Liu, Yuping Liu, Chao Li","doi":"10.1016/j.fsi.2024.109993","DOIUrl":"10.1016/j.fsi.2024.109993","url":null,"abstract":"<p><p>As a commercially valuable fish species, Sebastes schlegelii faces threats from pathogenic bacteria like Edwardsiella piscicida during aquaculture. The global host range of E. piscicida encompasses various species, yet its pathogenic mechanism remains incompletely elucidated. Cell lines offer invaluable in vitro resources for studying the pathogen pathogenicity. Here, we established and characterized a cell line derived from the intestinal tissue of the S. schlegelii, designated as SSI. SSI has undergone continuous subculturing for over 80 passages, demonstrating robust growth in DMEM supplemented with 10%-20% FBS and 20 μM HEPES at 24°C. Karyotype analysis and 18S rRNA amplification confirm its origin. SSI exhibits high transfection efficiency for exogenous DNA, making it suitable for gene expression and intestinal function analysis. E. piscicida infects SSI cells at low densities without inducing morphological changes within 6 h of infection, suggesting the potential of SSI as an in vitro model for studying E. piscicida pathogenicity. This cell line provides a valuable tool for investigating mucosal immunity and E. piscicida pathogenic mechanisms in marine fish.</p>","PeriodicalId":12127,"journal":{"name":"Fish & shellfish immunology","volume":" ","pages":"109993"},"PeriodicalIF":4.1,"publicationDate":"2024-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142557438","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 : 2024-11-01DOI: 10.1016/j.fsi.2024.109994
Carlo C. Lazado , Nora Albaladejo-Riad , Alexander Rebl
The internal timekeeping system regulates the daily cycle of physiological and behavioural changes in living organisms. This rhythmic phenomenon also influences cellular responses to reactive oxygen species, such as hydrogen peroxide (H2O2). However, the temporal interaction between H2O2 and fish mucosal cells is not well understood. This study examined the temporal variations of immunological and physiological responses to H2O2 in salmonid gill cells using the RTgill-W1 cell line. The results showed that gene expression levels varied during a 24-h cycle but did not exhibit rhythmicity. The presence of a 12-h light-dark cycle (12L:12D) signal increased gene expression levels compared to a 24-h dark cycle (0L:24D). To investigate whether the time of day affects the defences in gills, cells were exposed to H2O2 at two different times (Zeitgebertime 2, ZT2, or ZT14). Although significant expression changes were observed in genes related to stress and NF-κB signalling, only a limited time-dependent pattern of response to H2O2 was observed. The intracellular metabolome of gill cells was primarily composed of organic acid and derivatives, organoheterocyclic compounds, benzoids, organic oxygen and nitrogen compounds. Exposure to H2O2 at ZT2 led to significant changes in the metabolome compared to the control group, while no such changes were observed at ZT14. Within the control groups, the concentrations of 11 metabolites significantly varied between ZT2 and ZT14, with higher levels at ZT14. These metabolites were involved in arginine biosynthesis, amino acid metabolism, and nitrogen metabolism. In contrast, the level of 26 metabolites significantly varied between ZT2 and ZT14 in H2O2-exposed groups, with lower levels at ZT14. Comparing control and H2O2-exposed groups at ZT2, 38 metabolites were affected, primarily organic acid and derivatives and organic oxygen compounds. Functional annotation revealed that these altered metabolites were involved in 15 different pathways, with valine, leucine, and isoleucine biosynthesis being the most affected. This study reveals the presence of a time-dependent response to H2O2 in salmonid gill cells, which is reflected in the intracellular metabolome. The findings provide new insights into the temporal regulation of mucosal defences in fish.
{"title":"Intracellular metabolome elucidates the time-of-day-dependent response to hydrogen peroxide in salmonid gill epithelial cells","authors":"Carlo C. Lazado , Nora Albaladejo-Riad , Alexander Rebl","doi":"10.1016/j.fsi.2024.109994","DOIUrl":"10.1016/j.fsi.2024.109994","url":null,"abstract":"<div><div>The internal timekeeping system regulates the daily cycle of physiological and behavioural changes in living organisms. This rhythmic phenomenon also influences cellular responses to reactive oxygen species, such as hydrogen peroxide (H<sub>2</sub>O<sub>2</sub>). However, the temporal interaction between H<sub>2</sub>O<sub>2</sub> and fish mucosal cells is not well understood. This study examined the temporal variations of immunological and physiological responses to H<sub>2</sub>O<sub>2</sub> in salmonid gill cells using the RTgill-W1 cell line. The results showed that gene expression levels varied during a 24-h cycle but did not exhibit rhythmicity. The presence of a 12-h light-dark cycle (12L:12D) signal increased gene expression levels compared to a 24-h dark cycle (0L:24D). To investigate whether the time of day affects the defences in gills, cells were exposed to H<sub>2</sub>O<sub>2</sub> at two different times (<em>Zeitgeber</em> <em>time</em> 2, ZT2, or ZT14). Although significant expression changes were observed in genes related to stress and NF-κB signalling, only a limited time-dependent pattern of response to H<sub>2</sub>O<sub>2</sub> was observed. The intracellular metabolome of gill cells was primarily composed of organic acid and derivatives, organoheterocyclic compounds, benzoids, organic oxygen and nitrogen compounds. Exposure to H<sub>2</sub>O<sub>2</sub> at ZT2 led to significant changes in the metabolome compared to the control group, while no such changes were observed at ZT14. Within the control groups, the concentrations of 11 metabolites significantly varied between ZT2 and ZT14, with higher levels at ZT14. These metabolites were involved in arginine biosynthesis, amino acid metabolism, and nitrogen metabolism. In contrast, the level of 26 metabolites significantly varied between ZT2 and ZT14 in H<sub>2</sub>O<sub>2</sub>-exposed groups, with lower levels at ZT14. Comparing control and H<sub>2</sub>O<sub>2</sub>-exposed groups at ZT2, 38 metabolites were affected, primarily organic acid and derivatives and organic oxygen compounds. Functional annotation revealed that these altered metabolites were involved in 15 different pathways, with valine, leucine, and isoleucine biosynthesis being the most affected. This study reveals the presence of a time-dependent response to H<sub>2</sub>O<sub>2</sub> in salmonid gill cells, which is reflected in the intracellular metabolome. The findings provide new insights into the temporal regulation of mucosal defences in fish.</div></div>","PeriodicalId":12127,"journal":{"name":"Fish & shellfish immunology","volume":"154 ","pages":"Article 109994"},"PeriodicalIF":4.1,"publicationDate":"2024-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142557453","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
The live attenuated vaccine (P7-P8 strain) against cyprinid herpesvirus 2 (CyHV-2) infection of goldfish shows high protective efficacy. However, the underlying immune mechanism induced by P7-P8 vaccination remains unknown. It is known that the fish survived in the primary infection with CyHV-2 by the virus non-permissive high temperature (HT) water treatment elicit immunity against secondary virus challenge. In this study, the immunity induced by the P7-P8 vaccine was compared with that by HT treatment. To further explore the immunological responses of cyprinids, in addition to goldfish Carassius auratus, susceptible isogenic ginbuna C. auratus langsdorfii was included in this study. In the primary immune response, cyprinids were vaccinated with P7-P8, or treated with HT. In the secondary immune response, cyprinids were challenged with the virulent CyHV-2. The percentage dynamics of CD4-1 and CD8α positive lymphocytes were determined during the primary and secondary immune responses of the two cyprinids. Blood plasma was sampled to assess the anti-CyHV-2 IgM antibody titers. The vaccination with P7-P8 and HT-treatment induced high protection immunity in the cyprinids with relative percentage survival of over 88 % against virulent virus challenge. Our finding shows that the CD8α positive lymphocytes rather than CD4-1 positive lymphocytes play an important role in the secondary immune responses of cyprinids vaccinated with P7-P8. The CD4-1 positive lymphocytes rather than CD8α positive lymphocytes play an important role in the secondary immune responses of cyprinids treated with HT. The antibody titer of vaccinated cyprinids did not increase greatly even after virulent virus challenge. The results suggest the vaccine activates the CD8α cells and the secondary cell-mediated immunity. The differences in the induced immunity mechanisms in fish by the two measures might be based on the virus either being an avirulent virus form that cannot evade host responses or a virulent virus form that cannot propagate at non-permissive temperature.
{"title":"Cell-mediated and humoral immune responses of cyprinids induced by a live attenuated vaccine against cyprinid herpesvirus 2 infection in comparison to the virus non-permissive high temperature water treatment","authors":"Hiroaki Saito , Lik-Ming Lau , Shungo Minami , Manami Yuguchi , Megumi Matsumoto , Teruyuki Nakanishi , Hidehiro Kondo , Goshi Kato , Motohiko Sano","doi":"10.1016/j.fsi.2024.109991","DOIUrl":"10.1016/j.fsi.2024.109991","url":null,"abstract":"<div><div>The live attenuated vaccine (P7-P8 strain) against cyprinid herpesvirus 2 (CyHV-2) infection of goldfish shows high protective efficacy. However, the underlying immune mechanism induced by P7-P8 vaccination remains unknown. It is known that the fish survived in the primary infection with CyHV-2 by the virus non-permissive high temperature (HT) water treatment elicit immunity against secondary virus challenge. In this study, the immunity induced by the P7-P8 vaccine was compared with that by HT treatment. To further explore the immunological responses of cyprinids, in addition to goldfish <em>Carassius auratus</em>, susceptible isogenic ginbuna <em>C. auratus langsdorfii</em> was included in this study. In the primary immune response, cyprinids were vaccinated with P7-P8, or treated with HT. In the secondary immune response, cyprinids were challenged with the virulent CyHV-2. The percentage dynamics of CD4-1 and CD8α positive lymphocytes were determined during the primary and secondary immune responses of the two cyprinids. Blood plasma was sampled to assess the anti-CyHV-2 IgM antibody titers. The vaccination with P7-P8 and HT-treatment induced high protection immunity in the cyprinids with relative percentage survival of over 88 % against virulent virus challenge. Our finding shows that the CD8α positive lymphocytes rather than CD4-1 positive lymphocytes play an important role in the secondary immune responses of cyprinids vaccinated with P7-P8. The CD4-1 positive lymphocytes rather than CD8α positive lymphocytes play an important role in the secondary immune responses of cyprinids treated with HT. The antibody titer of vaccinated cyprinids did not increase greatly even after virulent virus challenge. The results suggest the vaccine activates the CD8α cells and the secondary cell-mediated immunity. The differences in the induced immunity mechanisms in fish by the two measures might be based on the virus either being an avirulent virus form that cannot evade host responses or a virulent virus form that cannot propagate at non-permissive temperature.</div></div>","PeriodicalId":12127,"journal":{"name":"Fish & shellfish immunology","volume":"154 ","pages":"Article 109991"},"PeriodicalIF":4.1,"publicationDate":"2024-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142544627","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 : 2024-11-01DOI: 10.1016/j.fsi.2024.109987
Chushan Dai , Yujia Miao , Zhan'ao Li , Yumian Liu , Ji Liu , Xiaoyu Liu , Shasha Tan , Hui Wu , Jun Xiao , Jun Zou , Hao Feng
RNF135, also known as RIPLET, plays a crucial role in facilitating RIG-I signaling in mammals. However, the function and regulatory mechanism of RNF135 in teleosts remain much to be elucidated. In this study, RNF135 homolog of black carp (bcRNF135) has been cloned and identified. The coding sequence (CDS) of bcRNF135 gene comprises 1221 nucleotides, encoding a protein of 407 amino acids. Immunoblotting (IB) and immunofluorescence (IF) assays identified that bcRNF135 is approximately 50 kDa and localized in the cytoplasm. qRT-PCR demonstrated that bcRNF135 mRNA levels were increased in host cells following SVCV infection and poly (I:C) stimulation. Co-expressed bcRNF135 obviously enhanced the induced transcription of IFN promoters by bcRIG-I in reporter assay, as well as improved bcRIG-I triggered antiviral response. Notably, bcRNF135 knockdown reduced the antiviral ability of host cells and increased virus replication. Co-immunoprecipitation (Co-IP) assays and IF assays confirmed that bcRNF135 interacted with bcRIG-I. Moreover, SDD-AGE revealed that bcRNF135 promotes the oligomerization of bcRIG-I, a process critical for RIG-I activation. Overall, our data conclude that bcRNF135 enhances bcRIG-I-mediated antiviral signaling by facilitating its ubiquitination and oligomerization, enriching our understanding of RIG-I regulation in teleost innate immunity.
{"title":"Black carp RNF135 enhances RIG-I-mediated antiviral signaling by facilitating its oligomerization","authors":"Chushan Dai , Yujia Miao , Zhan'ao Li , Yumian Liu , Ji Liu , Xiaoyu Liu , Shasha Tan , Hui Wu , Jun Xiao , Jun Zou , Hao Feng","doi":"10.1016/j.fsi.2024.109987","DOIUrl":"10.1016/j.fsi.2024.109987","url":null,"abstract":"<div><div>RNF135, also known as RIPLET, plays a crucial role in facilitating RIG-I signaling in mammals. However, the function and regulatory mechanism of RNF135 in teleosts remain much to be elucidated. In this study, RNF135 homolog of black carp (bcRNF135) has been cloned and identified. The coding sequence (CDS) of <em>bcRNF135</em> gene comprises 1221 nucleotides, encoding a protein of 407 amino acids. Immunoblotting (IB) and immunofluorescence (IF) assays identified that bcRNF135 is approximately 50 kDa and localized in the cytoplasm. qRT-PCR demonstrated that bcRNF135 mRNA levels were increased in host cells following SVCV infection and poly (I:C) stimulation. Co-expressed bcRNF135 obviously enhanced the induced transcription of IFN promoters by bcRIG-I in reporter assay, as well as improved bcRIG-I triggered antiviral response. Notably, bcRNF135 knockdown reduced the antiviral ability of host cells and increased virus replication. Co-immunoprecipitation (Co-IP) assays and IF assays confirmed that bcRNF135 interacted with bcRIG-I. Moreover, SDD-AGE revealed that bcRNF135 promotes the oligomerization of bcRIG-I, a process critical for RIG-I activation. Overall, our data conclude that bcRNF135 enhances bcRIG-I-mediated antiviral signaling by facilitating its ubiquitination and oligomerization, enriching our understanding of RIG-I regulation in teleost innate immunity.</div></div>","PeriodicalId":12127,"journal":{"name":"Fish & shellfish immunology","volume":"154 ","pages":"Article 109987"},"PeriodicalIF":4.1,"publicationDate":"2024-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142554339","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 : 2024-11-01DOI: 10.1016/j.fsi.2024.110000
Nguyen Giang Thu Lan, Ha Thanh Dong, Nguyen Tien Vinh, Krishna R Salin, Saengchan Senapin, Khaettareeya Pimsannil, Sophie St-Hilaire, Andrew P Shinn, Channarong Rodkhum
{"title":"Corrigendum to \"A novel vaccination strategy against Vibrio harveyi infection in Asian seabass (Lates calcarifer) with the aid of oxygen nanobubbles and chitosan\" [Fish and Shellfish Immunol 149 (2024) 109557].","authors":"Nguyen Giang Thu Lan, Ha Thanh Dong, Nguyen Tien Vinh, Krishna R Salin, Saengchan Senapin, Khaettareeya Pimsannil, Sophie St-Hilaire, Andrew P Shinn, Channarong Rodkhum","doi":"10.1016/j.fsi.2024.110000","DOIUrl":"10.1016/j.fsi.2024.110000","url":null,"abstract":"","PeriodicalId":12127,"journal":{"name":"Fish & shellfish immunology","volume":" ","pages":"110000"},"PeriodicalIF":4.1,"publicationDate":"2024-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142564203","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 : 2024-11-01DOI: 10.1016/j.fsi.2024.109983
Xianfa Zhou , Dongfang Sun , Junyang Guo , Jianjian Lv , Ping Liu , Baoquan Gao
Vibrio parahaemolyticus is the main pathogen causing acute hepatopancreatic necrotic disease in crustaceans. To elucidate the epigenetic regulatory mechanism of crustacean resistance to V. parahaemolyticus infection, we conducted artificial infection studies on Portunus trituberculatus. The results showed that the mortality rate reached the highest at 12 h of artificial infection, which was 23.69 %. At 72 h after V parahaemolyticus infection, the expression level of DNA demethylase (ten-eleven-translocation protein) Tet was significantly decreased, the expression of DNA methyltransferase Dnmt3B fluctuated significantly. Based on the differential expression levels of Tet and Dnmt3B. We depict for DNA methylation profiles of the whole genome of P. trituberculatus at single-base resolution by using whole-genome bisulfite sequencing (WGBS) on hemolymph tissues. The overall DNA methylation level was low at 2.16 % in P. trituberculatus hemolymph. A total of 2590 differentially methylated regions (DMRs) were identified, of which 1329 were hypermethylated and 1261 were hypomethylated, and 1389 genes were annotated in these DMRs. Differently methylated genes (DMGs) were significantly enriched in ribosomes (KO03010), protein kinases (KO01001), cell cycle (HSA04110), endocrine resistance (HSA01522) and FoxO signaling pathway (KO04068). Finally, we selected six differentially methylated genes for quantitative analysis. The results showed that DNA methylation not only has a negative regulatory effect on gene expression, but also has a positive regulatory effect. These results indicated that DNA methylation in the regulation of genes involved in immune responses contributes to the resistance of P. trituberculatus to V. parahaemolyticus, which is valuable for understanding how crustaceans regulate the innate immune system to defend against bacterial infections.
{"title":"Insights into the DNA methylation of Portunus trituberculatus in response to Vibrio parahaemolyticus infection","authors":"Xianfa Zhou , Dongfang Sun , Junyang Guo , Jianjian Lv , Ping Liu , Baoquan Gao","doi":"10.1016/j.fsi.2024.109983","DOIUrl":"10.1016/j.fsi.2024.109983","url":null,"abstract":"<div><div><em>Vibrio parahaemolyticus</em> is the main pathogen causing acute hepatopancreatic necrotic disease in crustaceans. To elucidate the epigenetic regulatory mechanism of crustacean resistance to <em>V</em>. <em>parahaemolyticus</em> infection, we conducted artificial infection studies on <em>Portunus trituberculatus</em>. The results showed that the mortality rate reached the highest at 12 h of artificial infection, which was 23.69 %. At 72 h after <em>V parahaemolyticus</em> infection, the expression level of DNA demethylase (ten-eleven-translocation protein) <em>Tet</em> was significantly decreased, the expression of DNA methyltransferase <em>Dnmt3B</em> fluctuated significantly. Based on the differential expression levels of <em>Tet</em> and <em>Dnmt3B</em>. We depict for DNA methylation profiles of the whole genome of <em>P. trituberculatus</em> at single-base resolution by using whole-genome bisulfite sequencing (WGBS) on hemolymph tissues. The overall DNA methylation level was low at 2.16 % in <em>P. trituberculatus</em> hemolymph. A total of 2590 differentially methylated regions (DMRs) were identified, of which 1329 were hypermethylated and 1261 were hypomethylated, and 1389 genes were annotated in these DMRs. Differently methylated genes (DMGs) were significantly enriched in ribosomes (KO03010), protein kinases (KO01001), cell cycle (HSA04110), endocrine resistance (HSA01522) and FoxO signaling pathway (KO04068). Finally, we selected six differentially methylated genes for quantitative analysis. The results showed that DNA methylation not only has a negative regulatory effect on gene expression, but also has a positive regulatory effect. These results indicated that DNA methylation in the regulation of genes involved in immune responses contributes to the resistance of <em>P</em>. <em>trituberculatus</em> to <em>V</em>. <em>parahaemolyticus</em>, which is valuable for understanding how crustaceans regulate the innate immune system to defend against bacterial infections.</div></div>","PeriodicalId":12127,"journal":{"name":"Fish & shellfish immunology","volume":"154 ","pages":"Article 109983"},"PeriodicalIF":4.1,"publicationDate":"2024-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142497577","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 : 2024-11-01DOI: 10.1016/j.fsi.2024.110001
Hong Jiang , Jiahui Zhang , Tingting Liu , Xinping Chen , Guiwen Yang , Hua Li
The NLRP1 inflammasome is a crucial muti-protein complex in the host anti-pathogen immune response. The previous studies have revealed that the anti-apoptotic protein BCL-xL played a non-apoptotic role by impeding the activation of NLRP1 inflammasome in mammals. However, the potential role of BCL-xL in regulating the inflammasome in fish remains unclear. In the present study, the BCL-xL (CcBCL-xL) was cloned from the head kidney of common carp (Cyprinus carpio L.), and its regulatory effect on the NLRP1 inflammasome was explored. It was found that CcBCL-xL predominantly localized in the brain, spleen and head kidney of common carp, and upon stimulation with Aeromonas hydrophila (A. hydrophila), Edwardsiella tarda (E. tarda), or spring viremia of carp virus (SVCV), the expression of CcBCL-xL significantly increased in multiple immune organs. The interaction between CcBCL-xL and CcNLRP1 was confirmed by co-immunoprecipitation and immunofluorescence. Meanwhile, we also found that CcBCL-xL significantly inhibited the assembly of the CcNLRP1 inflammasome, through ASC oligomerization, ASC specks formation and cytotoxicity experiments. Furthermore, our results revealed that CcBCL-xL interacted with the NACHT, LRR, FIIND, and CARD domains of CcNLRP1. Taken together, the results provide a theoretical foundation for further exploring the regulatory mechanism of NLRP1, and for the prevention and treatment of infectious diseases in fish.
{"title":"The characterization of BCL-xL displays a non-apoptotic role in suppression of NLRP1 inflammasome assembly in common carp (Cyprinus carpio L.)","authors":"Hong Jiang , Jiahui Zhang , Tingting Liu , Xinping Chen , Guiwen Yang , Hua Li","doi":"10.1016/j.fsi.2024.110001","DOIUrl":"10.1016/j.fsi.2024.110001","url":null,"abstract":"<div><div>The NLRP1 inflammasome is a crucial muti-protein complex in the host anti-pathogen immune response. The previous studies have revealed that the anti-apoptotic protein BCL-xL played a non-apoptotic role by impeding the activation of NLRP1 inflammasome in mammals. However, the potential role of BCL-xL in regulating the inflammasome in fish remains unclear. In the present study, the BCL-xL (<em>Cc</em>BCL-xL) was cloned from the head kidney of common carp (<em>Cyprinus carpio</em> L.), and its regulatory effect on the NLRP1 inflammasome was explored. It was found that <em>Cc</em>BCL-xL predominantly localized in the brain, spleen and head kidney of common carp, and upon stimulation with <em>Aeromonas hydrophila</em> (<em>A. hydrophila</em>), <em>Edwardsiella tarda</em> (<em>E. tarda</em>), or spring viremia of carp virus (SVCV), the expression of <em>Cc</em>BCL-xL significantly increased in multiple immune organs. The interaction between <em>Cc</em>BCL-xL and <em>Cc</em>NLRP1 was confirmed by co-immunoprecipitation and immunofluorescence. Meanwhile, we also found that <em>Cc</em>BCL-xL significantly inhibited the assembly of the <em>Cc</em>NLRP1 inflammasome, through ASC oligomerization, ASC specks formation and cytotoxicity experiments. Furthermore, our results revealed that <em>Cc</em>BCL-xL interacted with the NACHT, LRR, FIIND, and CARD domains of <em>Cc</em>NLRP1. Taken together, the results provide a theoretical foundation for further exploring the regulatory mechanism of NLRP1, and for the prevention and treatment of infectious diseases in fish.</div></div>","PeriodicalId":12127,"journal":{"name":"Fish & shellfish immunology","volume":"155 ","pages":"Article 110001"},"PeriodicalIF":4.1,"publicationDate":"2024-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142568267","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 : 2024-10-31DOI: 10.1016/j.fsi.2024.109997
Xinli Ding , Kaini Zhang , Qianmin Zhuang , Yanru Chen , Hua Li , Shili Liu , Lei Chen
Objectives
Peptidoglycan recognition protein 2 (PGRP2) plays a role in regulating immune defense in fish. Our previous studies found that CcPGRP2 helped maintain the integrity of the intestinal mucosa of carp and could bind and agglutinate bacteria when infected with A. hydrophila. However, its effect on the structure of the microbiota has not yet been clarified. Therefore, it is necessary to explore the effect of CcPGRP2 on the intestinal microbiota structure in fish.
Methods
In the present study, common carp were injected with CcPGRP2 protein intraperitoneally and high-throughput sequencing technology was used to study the difference in intestinal microbiota structure. Firstly, the variations in α- and β-diversity of the intestinal microbiota of common carp in control and treatment groups were tested, and the results indicated that intraperitoneal injection of A. hydrophila significantly reduced the microbial α-diversity (within-samples) and β-diversity (between-samples) in common carp gut samples, but CcPGRP2 protein could alleviate these reduction, no matter in the case of simultaneous injection of CcPGRP2 protein and A. hydrophila or a intermitted injection with first injection of CcPGRP2 and then A. hydrophila after 6 h. Subsequently, the intestinal microbiota structures of common carp on various taxonomic levels were interrogated under the treatments.
Results
The data revealed that the abundance of intestinal pathogen Aeromonas was reduced when CcPGRP2 was injected in the common carp, and the alleviation effect was better when CcPGRP2 was injected with A. hydrophila at the same time, implying the function of CcPGRP2 in inhibiting intestinal dysbiosis. Moreover, the functional prediction demonstrated the possible physiological shifts and the influences of microbes on the environment after the common carp is injected with A. hydrophila and CcPGRP2. Finally, the bacterial interaction patterns results showed that the groups injected with A. hydrophila were diverted away from the control group in terms of clustering relationship, while the injection of CcPGRP2 could reverse the effect of A. hydrophila and keep the microbial structure closer to that of the control group; meanwhile, the effect of simultaneous injection of A. hydrophila and CcPGRP2 was better than that of intermitted injections.
Conclusions
All the results in this study suggest that the CcPGRP2 could alleviate the internal dysbiosis under pathogen infection, which will provide a foundation for disease resistance breeding.
{"title":"Common carp Peptidoglycan Gecognition Protein 2 (CcPGRP2) alleviates gut dysbiosis induced by Aeromonas hydrophila","authors":"Xinli Ding , Kaini Zhang , Qianmin Zhuang , Yanru Chen , Hua Li , Shili Liu , Lei Chen","doi":"10.1016/j.fsi.2024.109997","DOIUrl":"10.1016/j.fsi.2024.109997","url":null,"abstract":"<div><h3>Objectives</h3><div>Peptidoglycan recognition protein 2 (PGRP2) plays a role in regulating immune defense in fish. Our previous studies found that CcPGRP2 helped maintain the integrity of the intestinal mucosa of carp and could bind and agglutinate bacteria when infected with <em>A. hydrophila</em>. However, its effect on the structure of the microbiota has not yet been clarified. Therefore, it is necessary to explore the effect of CcPGRP2 on the intestinal microbiota structure in fish.</div></div><div><h3>Methods</h3><div>In the present study, common carp were injected with CcPGRP2 protein intraperitoneally and high-throughput sequencing technology was used to study the difference in intestinal microbiota structure. Firstly, the variations in α- and β-diversity of the intestinal microbiota of common carp in control and treatment groups were tested, and the results indicated that intraperitoneal injection of <em>A. hydrophila</em> significantly reduced the microbial α-diversity (within-samples) and β-diversity (between-samples) in common carp gut samples, but CcPGRP2 protein could alleviate these reduction, no matter in the case of simultaneous injection of CcPGRP2 protein and <em>A. hydrophila</em> or a intermitted injection with first injection of CcPGRP2 and then <em>A. hydrophila</em> after 6 h. Subsequently, the intestinal microbiota structures of common carp on various taxonomic levels were interrogated under the treatments.</div></div><div><h3>Results</h3><div>The data revealed that the abundance of intestinal pathogen <em>Aeromonas</em> was reduced when CcPGRP2 was injected in the common carp, and the alleviation effect was better when CcPGRP2 was injected with <em>A. hydrophila</em> at the same time, implying the function of CcPGRP2 in inhibiting intestinal dysbiosis. Moreover, the functional prediction demonstrated the possible physiological shifts and the influences of microbes on the environment after the common carp is injected with <em>A. hydrophila</em> and CcPGRP2. Finally, the bacterial interaction patterns results showed that the groups injected with <em>A. hydrophila</em> were diverted away from the control group in terms of clustering relationship, while the injection of CcPGRP2 could reverse the effect of <em>A. hydrophila</em> and keep the microbial structure closer to that of the control group; meanwhile, the effect of simultaneous injection of <em>A. hydrophila</em> and CcPGRP2 was better than that of intermitted injections.</div></div><div><h3>Conclusions</h3><div>All the results in this study suggest that the CcPGRP2 could alleviate the internal dysbiosis under pathogen infection, which will provide a foundation for disease resistance breeding.</div></div>","PeriodicalId":12127,"journal":{"name":"Fish & shellfish immunology","volume":"155 ","pages":"Article 109997"},"PeriodicalIF":4.1,"publicationDate":"2024-10-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142564196","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 : 2024-10-31DOI: 10.1016/j.fsi.2024.109999
Xuangang Wang , Xiangfu Kong , Zhentao Chen , Hengshun Li , Ze Tao , Quanqi Zhang , Haiyang Yu
Sebastes schlegelii is an economically significant marine fish that faces serious threats from various pathogens. Edwardsiella piscicida is a pathogenic bacterium that primarily affects fish, including S. schlegelii, leading to severe disease. Although numerous reports have documented the transcriptome sequencing of various fish tissues in response to E. piscicida infection, studies focusing on specific cells remain scarce. In this study, S. schlegelii were infected by intraperitoneal injection of E. piscicida. Severe external clinical signs were observed in E. piscicida-infected S. schlegelii and pathological examination demonstrated structural damage of the head kidney following treatment with E. piscicida. Furthermore, macrophages were isolated from the head kidneys of both the control and E. piscicida-infected groups for RNA sequencing (RNA-seq). Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis indicated that the DEGs were closely associated with immune response and oxidative stress. Additionally, Weighted Gene Co-expression Network Analysis (WGCNA) was performed based on the data from this study and RNA-seq files of macrophages infected with E. piscicida in vitro, revealing that immune responses, oxidative stress, and mitochondrial damage were involved in the macrophage response to E. piscicida infection both in vivo and in vitro. This study provides a reference for understanding the mechanisms by which teleost immune cells respond to pathogen invasion and enhances our comprehension of teleost innate immunity.
石首鱼是一种具有重要经济价值的海水鱼，面临着各种病原体的严重威胁。Edwardsiella piscicida 是一种致病细菌，主要影响鱼类，包括 S. schlegelii，导致严重疾病。尽管许多报告都记录了各种鱼类组织对 E. piscicida 感染反应的转录组测序，但针对特定细胞的研究仍然很少。在本研究中，通过腹腔注射 E. piscicida 感染了 S. schlegelii。经 E. piscicida 感染的 S. schlegelii 出现了严重的外部临床症状，病理检查显示，经 E. piscicida 治疗后，头部肾脏结构受损。此外，从对照组和E. piscicida感染组的头肾中分离出巨噬细胞，进行RNA测序（RNA-seq）。基因本体（GO）和京都基因组百科全书（KEGG）分析表明，DEGs与免疫反应和氧化应激密切相关。此外，根据本研究的数据和体外感染鱼腥藻的巨噬细胞的RNA-seq文件，进行了加权基因共表达网络分析（WGCNA），发现免疫反应、氧化应激和线粒体损伤参与了巨噬细胞对体内和体外鱼腥藻感染的反应。这项研究为了解远洋渔业免疫细胞应对病原体入侵的机制提供了参考，并加深了我们对远洋渔业先天免疫的理解。
{"title":"Transcriptome analysis reveals the mechanism of black rockfish (Sebastes schlegelii) macrophages respond to Edwardsiella piscicida infection in vivo","authors":"Xuangang Wang , Xiangfu Kong , Zhentao Chen , Hengshun Li , Ze Tao , Quanqi Zhang , Haiyang Yu","doi":"10.1016/j.fsi.2024.109999","DOIUrl":"10.1016/j.fsi.2024.109999","url":null,"abstract":"<div><div><em>Sebastes schlegelii</em> is an economically significant marine fish that faces serious threats from various pathogens. <em>Edwardsiella piscicida</em> is a pathogenic bacterium that primarily affects fish, including <em>S</em>. <em>schlegelii</em>, leading to severe disease. Although numerous reports have documented the transcriptome sequencing of various fish tissues in response to <em>E. piscicida</em> infection, studies focusing on specific cells remain scarce. In this study, <em>S</em>. <em>schlegelii</em> were infected by intraperitoneal injection of <em>E. piscicida</em>. Severe external clinical signs were observed in <em>E. piscicida</em>-infected <em>S</em>. <em>schlegelii</em> and pathological examination demonstrated structural damage of the head kidney following treatment with <em>E. piscicida</em>. Furthermore, macrophages were isolated from the head kidneys of both the control and <em>E. piscicida</em>-infected groups for RNA sequencing (RNA-seq). Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis indicated that the DEGs were closely associated with immune response and oxidative stress. Additionally, Weighted Gene Co-expression Network Analysis (WGCNA) was performed based on the data from this study and RNA-seq files of macrophages infected with <em>E. piscicida</em> in vitro, revealing that immune responses, oxidative stress, and mitochondrial damage were involved in the macrophage response to <em>E. piscicida</em> infection both in vivo and in vitro. This study provides a reference for understanding the mechanisms by which teleost immune cells respond to pathogen invasion and enhances our comprehension of teleost innate immunity.</div></div>","PeriodicalId":12127,"journal":{"name":"Fish & shellfish immunology","volume":"155 ","pages":"Article 109999"},"PeriodicalIF":4.1,"publicationDate":"2024-10-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142564210","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 : 2024-10-30DOI: 10.1016/j.fsi.2024.109995
Yi Zhang , Xiaoli Zhang , Jiaxin Xu , Jinbin Zheng , Zhaoxia Cui
Water pH is a critical environmental factor for aquaculture. Acidification is a pressing environmental issue that poses significant threats to the aquaculture industry. Since the outbreaks of disease generally accompany with environmental stress, comparative transcriptome analyses were performed to investigate the combined effects of low pH stress and bacterial infection on the transcriptional changes of hemocytes in the economically important crab Eriocheir sinensis. The results revealed that the immune deficiency (IMD) pathway and prophenoloxidase (proPO) system was activated to defense against Vibro parahaemolyticus even when crabs were subjected to low pH stress, whereas low pH stress resulted in the disorder of Toll-like receptor (TLR) pathway upon V. parahaemolyticus infection. Moreover, low pH stress might weaken crabs’ defense against V. parahaemolyticus by inhibiting the up-regulation of crustin and suppressing the expression of lysozyme, and disturb the maintaining of protein homeostasis through the transcriptional decrement of a batch of heat shock proteins (HSPs). It is worth noting that both V. parahaemolyticus infection and low pH stress might suppress the energy metabolism in the hemocytes via inhibiting the expression of critical enzymes, dihydrolipoyllysine-residue acetyltransferase component of pyruvate dehydrogenase complex and fumarase, in the tricarboxylic acid (TCA) cycle. This study provides novel understandings concerning the transcriptional changes of hemocyte in E. sinensis subjected to a combination of low pH stress and V. parahaemolyticus infection as well as contribute to optimize the management strategies for the prevention and control of diseases in E. sinensis farming.
{"title":"Combined effects of low pH stress and bacterial infection on the transcriptional changes of hemocytes in Chinese mitten crab Eriocheir sinensis","authors":"Yi Zhang , Xiaoli Zhang , Jiaxin Xu , Jinbin Zheng , Zhaoxia Cui","doi":"10.1016/j.fsi.2024.109995","DOIUrl":"10.1016/j.fsi.2024.109995","url":null,"abstract":"<div><div>Water pH is a critical environmental factor for aquaculture. Acidification is a pressing environmental issue that poses significant threats to the aquaculture industry. Since the outbreaks of disease generally accompany with environmental stress, comparative transcriptome analyses were performed to investigate the combined effects of low pH stress and bacterial infection on the transcriptional changes of hemocytes in the economically important crab <em>Eriocheir sinensis</em>. The results revealed that the immune deficiency (IMD) pathway and prophenoloxidase (proPO) system was activated to defense against <em>Vibro parahaemolyticus</em> even when crabs were subjected to low pH stress, whereas low pH stress resulted in the disorder of Toll-like receptor (TLR) pathway upon <em>V</em>. <em>parahaemolyticus</em> infection. Moreover, low pH stress might weaken crabs’ defense against <em>V</em>. <em>parahaemolyticus</em> by inhibiting the up-regulation of crustin and suppressing the expression of lysozyme, and disturb the maintaining of protein homeostasis through the transcriptional decrement of a batch of heat shock proteins (HSPs). It is worth noting that both <em>V</em>. <em>parahaemolyticus</em> infection and low pH stress might suppress the energy metabolism in the hemocytes via inhibiting the expression of critical enzymes, dihydrolipoyllysine-residue acetyltransferase component of pyruvate dehydrogenase complex and fumarase, in the tricarboxylic acid (TCA) cycle. This study provides novel understandings concerning the transcriptional changes of hemocyte in <em>E</em>. <em>sinensis</em> subjected to a combination of low pH stress and <em>V</em>. <em>parahaemolyticus</em> infection as well as contribute to optimize the management strategies for the prevention and control of diseases in <em>E</em>. <em>sinensis</em> farming.</div></div>","PeriodicalId":12127,"journal":{"name":"Fish & shellfish immunology","volume":"155 ","pages":"Article 109995"},"PeriodicalIF":4.1,"publicationDate":"2024-10-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142557437","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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