- Email: [email protected]
Atlantic salmon interferon receptors: Gene clusters and expression
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Abstracts / Fish & Shellfish Immunology 34 (2013) 1692–1752
* Corresponding author.
P-326.
E-mail address: [email protected] (M.K. Raida)
Atlantic salmon interferon receptors: Gene clusters and expression B. Sun 1, B. Koop 2, B. Robertsen 1, *.
P-334. The immune response of giant freshwater prawn (Macrobrachium resenbergii de Man) against Lactococcus garvieae infection N. Suanyuk*, M. Dangwetngam, C. Tantikitti. Kidchakan Supamattaya Aquatic Animal Health Research Center, Department of Aquatic Science, Faculty of Natural Resources, Prince of Songkla University, Hat Yai, Songkhla 90112, Thailand Abstract Bacterial diseases caused by Lactococcus garvieae becoming an increasing problem in aquaculture. Between 2009 and 2011, this bacteria was isolated from giant freshwater prawn (Macrobrachium rosenbergii de Man) from both cultured and naturally environments in Patthalung and Songkhla provinces, southern Thailand. Identifications by conventional and rapid identification systems, as well as genetic and phylogenetic characterization indicated that the bacteria was L. garvieae. Analysis of 16S rRNA revealed 100% homology to the 16S rRNA sequence of several L. garvieae strains in GenBank. Studies on hemato-immunological parameters of giant freshwater prawn infected with L. garvieae showed the reduction in total hemocyte count, phenoloxidase activity, ratio of oxyhemocyanin to protein and glucose level. Hemolymph protein was in the similar range among L. garvieae-infected and noninfected prawn. The present study indicated that L. garvieae caused depression in the immune response of giant freshwater prawn. * Corresponding author. E-mail address: [email protected] (N. Suanyuk)
P-221. Identification and expression analysis of IFN-gamma related (IFN-grel) genes in Atlantic salmon Salmo salar T. Summathed 1, J. Zou 1, *, S. Wadsworth 2, C. Secombes 1.
1
Norwegian College of Fishery Science, University of Tromsø, 9037 Tromsø, Norway; 2 Centre for Biomedical Research, University of Victoria, BC, Canada Abstract In mammals, type I interferons (IFNs) all mediate antiviral responses by binding to a common receptor complex containing two chains of IFNAR1 and IFNAR2, which belong to the class II cytokine receptor family (CRFB). In zebrafish, it has been suggested that type I IFNs bind to two distinct receptor complexes: group I IFNs with 2 cysteines bind to receptor complex containing CRFB1 and CRFB5 while group II IFNs with 4 cysteines bind to receptor complex of CRFB2 and CRFB5. The CRFB gene clusters in Atlantic salmon seem to be more complicated than in zebrafish. In this work we identified two CRFB1, one CRFB2 and six CRFB5 genes. CRFB1a is a soluble CRFB without transmembrane region and CRFB1b and CRFB2 are intact receptor side chains. CRFB5a, CRFB5b, CRFB5c and CRFB5d are intact receptor subunits, while CRFB5t has a short transmembrane, and CRFB5s is a soluble CRFB without transmembrane region. In mammals, IFNAR transcripts encoding three isoforms are generated from the same gene by exon skipping, alternative splicing, and differential usage of polyadenylation sites while in salmon the soluble form and truncated form CRFBs are encoded by separate genes. The expression of different CRFBs in TO cells after IPN virus or ISA virus was investigated, and it is interesting to find out soluble CRFB1a could be induced by both virus. Gene transcription of CRFBs were examined in different tissues: head kidney, kidney, spleen, heart, intestine, brain, ovary, muscle, gill and liver. The mRNA of two high affinity binding chains CRFB1b and CRFB2 have preferred tissue distribution. There are more CRFB2 mRNA in heart and ovary, while CRFB1b is the dominant affinity biding chain in intestine and muscle. The expression of CRFBs were also investigated in isolated head kidney leucocytes after poly I:C or imidazoquinoline R848 stimulation. * Corresponding author. E-mail address: [email protected] (B. Robertsen)
1
Scottish Fish Immunology Research Centre, Institute of Biological and Environmental Sciences, University of Aberdeen, Tillydrone Avenue, Aberdeen AB24 2TZ, UK; 2 EWOS Innovation A.S., Dirdal, N-4335, Norway Abstract Atlantic salmon (Salmo salar L.) is the most important commercial species in the Scottish aquaculture industry. Many viral diseases can impact on the industry, leading to economic losses. Interferons (IFNs) are cytokines involved in antiviral defence, and study of their regulation and function may shed light on how fish can defend themselves against virus attack. Two types exist in fish, type I and type II. The latter include two members in teleost fish, IFN-g and IFN-grel. In this study, the IFN-grel genes have been characterized in salmonids for the first time, by in silico analysis of the whole genome shotgun sequence database of Atlantic salmon and subsequent confirmation by cloning from the salmon head kidney cell line SHK-1. The salmon IFN-grel proteins share low sequence homology with IFN-g and can be divided into two phylogenetic groups. Homology protein structure modelling revealed that both IFN-g and IFN-grel are alpha-helical proteins similar to human IFN-g, but have some major structural differences at the N- and C-terminal region, which in human IFN-g is known to be involved in receptor binding. Putative binding sites for transcription factors such as Tbet, STAT, IRF9, NF-kB and NFAT are found in the IFN-grel promoter. Expression analysis found upregulation of IFN-grel gene in PHA treated SHK-1 cells and primary head kidney leukocytes. The regulation of IFN-grel gene expression by antiviral and proinflammatory cytokines will be further analysed. * Corresponding author. E-mail address: [email protected] (J. Zou)
P-225. Transcriptomic analysis on the initiation of acute WSSV infection caused by temperature change Y. Sun, F. Li*, Z. Sun, J. Xiang*. Key Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences, Qingdao 266071, China Abstract White spot syndrome virus (WSSV) is the most severe epidemic pathogen to the shrimp culture industry worldwide. In order to understand the biological process of the host at the initial stage of the rapid propagation of WSSV and the initiation mechanisms of the acute infection caused by temperature rising shortly from low temperature (18 C) to high temperature (25 C), RNA-Seq was used to analyze the differential transcriptional expressions between challenged shrimp cultured at 18 C and that cultured at 25 C. Three groups of Exopalamon carincauda Holthuis, respectively named as Group T, Group W and Group WT were set. Shrimp in Group T were cultured at 18 C without any treatment; Shrimp in Group W and Group WT were cultured at 18 C and then injected with WSSV. At 24 hours post challenge, the water temperature of Group T and Group WT was increased to 25 C. Cephalothorax samples were collected from Group W and Group WT at 12 hours post temperature rising and RNA-seq was performed. Totally 61,886 unigenes with an average length of 626 bp were obtained, among which about 6,421 were differentially expressed genes (DEGs). 4689 DEGs were down-regulated in Group WT and the other 1,732 DEGs were up-regulated, of which 173 were WSSV genes. According to the Gene Ontology analysis, the DEGs were involved in various biological
Abstracts / Fish & Shellfish Immunology 34 (2013) 1692–1752
* Corresponding author.
P-326.
E-mail address: [email protected] (M.K. Raida)
Atlantic salmon interferon receptors: Gene clusters and expression B. Sun 1, B. Koop 2, B. Robertsen 1, *.
P-334. The immune response of giant freshwater prawn (Macrobrachium resenbergii de Man) against Lactococcus garvieae infection N. Suanyuk*, M. Dangwetngam, C. Tantikitti. Kidchakan Supamattaya Aquatic Animal Health Research Center, Department of Aquatic Science, Faculty of Natural Resources, Prince of Songkla University, Hat Yai, Songkhla 90112, Thailand Abstract Bacterial diseases caused by Lactococcus garvieae becoming an increasing problem in aquaculture. Between 2009 and 2011, this bacteria was isolated from giant freshwater prawn (Macrobrachium rosenbergii de Man) from both cultured and naturally environments in Patthalung and Songkhla provinces, southern Thailand. Identifications by conventional and rapid identification systems, as well as genetic and phylogenetic characterization indicated that the bacteria was L. garvieae. Analysis of 16S rRNA revealed 100% homology to the 16S rRNA sequence of several L. garvieae strains in GenBank. Studies on hemato-immunological parameters of giant freshwater prawn infected with L. garvieae showed the reduction in total hemocyte count, phenoloxidase activity, ratio of oxyhemocyanin to protein and glucose level. Hemolymph protein was in the similar range among L. garvieae-infected and noninfected prawn. The present study indicated that L. garvieae caused depression in the immune response of giant freshwater prawn. * Corresponding author. E-mail address: [email protected] (N. Suanyuk)
P-221. Identification and expression analysis of IFN-gamma related (IFN-grel) genes in Atlantic salmon Salmo salar T. Summathed 1, J. Zou 1, *, S. Wadsworth 2, C. Secombes 1.
1
Norwegian College of Fishery Science, University of Tromsø, 9037 Tromsø, Norway; 2 Centre for Biomedical Research, University of Victoria, BC, Canada Abstract In mammals, type I interferons (IFNs) all mediate antiviral responses by binding to a common receptor complex containing two chains of IFNAR1 and IFNAR2, which belong to the class II cytokine receptor family (CRFB). In zebrafish, it has been suggested that type I IFNs bind to two distinct receptor complexes: group I IFNs with 2 cysteines bind to receptor complex containing CRFB1 and CRFB5 while group II IFNs with 4 cysteines bind to receptor complex of CRFB2 and CRFB5. The CRFB gene clusters in Atlantic salmon seem to be more complicated than in zebrafish. In this work we identified two CRFB1, one CRFB2 and six CRFB5 genes. CRFB1a is a soluble CRFB without transmembrane region and CRFB1b and CRFB2 are intact receptor side chains. CRFB5a, CRFB5b, CRFB5c and CRFB5d are intact receptor subunits, while CRFB5t has a short transmembrane, and CRFB5s is a soluble CRFB without transmembrane region. In mammals, IFNAR transcripts encoding three isoforms are generated from the same gene by exon skipping, alternative splicing, and differential usage of polyadenylation sites while in salmon the soluble form and truncated form CRFBs are encoded by separate genes. The expression of different CRFBs in TO cells after IPN virus or ISA virus was investigated, and it is interesting to find out soluble CRFB1a could be induced by both virus. Gene transcription of CRFBs were examined in different tissues: head kidney, kidney, spleen, heart, intestine, brain, ovary, muscle, gill and liver. The mRNA of two high affinity binding chains CRFB1b and CRFB2 have preferred tissue distribution. There are more CRFB2 mRNA in heart and ovary, while CRFB1b is the dominant affinity biding chain in intestine and muscle. The expression of CRFBs were also investigated in isolated head kidney leucocytes after poly I:C or imidazoquinoline R848 stimulation. * Corresponding author. E-mail address: [email protected] (B. Robertsen)
1
Scottish Fish Immunology Research Centre, Institute of Biological and Environmental Sciences, University of Aberdeen, Tillydrone Avenue, Aberdeen AB24 2TZ, UK; 2 EWOS Innovation A.S., Dirdal, N-4335, Norway Abstract Atlantic salmon (Salmo salar L.) is the most important commercial species in the Scottish aquaculture industry. Many viral diseases can impact on the industry, leading to economic losses. Interferons (IFNs) are cytokines involved in antiviral defence, and study of their regulation and function may shed light on how fish can defend themselves against virus attack. Two types exist in fish, type I and type II. The latter include two members in teleost fish, IFN-g and IFN-grel. In this study, the IFN-grel genes have been characterized in salmonids for the first time, by in silico analysis of the whole genome shotgun sequence database of Atlantic salmon and subsequent confirmation by cloning from the salmon head kidney cell line SHK-1. The salmon IFN-grel proteins share low sequence homology with IFN-g and can be divided into two phylogenetic groups. Homology protein structure modelling revealed that both IFN-g and IFN-grel are alpha-helical proteins similar to human IFN-g, but have some major structural differences at the N- and C-terminal region, which in human IFN-g is known to be involved in receptor binding. Putative binding sites for transcription factors such as Tbet, STAT, IRF9, NF-kB and NFAT are found in the IFN-grel promoter. Expression analysis found upregulation of IFN-grel gene in PHA treated SHK-1 cells and primary head kidney leukocytes. The regulation of IFN-grel gene expression by antiviral and proinflammatory cytokines will be further analysed. * Corresponding author. E-mail address: [email protected] (J. Zou)
P-225. Transcriptomic analysis on the initiation of acute WSSV infection caused by temperature change Y. Sun, F. Li*, Z. Sun, J. Xiang*. Key Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences, Qingdao 266071, China Abstract White spot syndrome virus (WSSV) is the most severe epidemic pathogen to the shrimp culture industry worldwide. In order to understand the biological process of the host at the initial stage of the rapid propagation of WSSV and the initiation mechanisms of the acute infection caused by temperature rising shortly from low temperature (18 C) to high temperature (25 C), RNA-Seq was used to analyze the differential transcriptional expressions between challenged shrimp cultured at 18 C and that cultured at 25 C. Three groups of Exopalamon carincauda Holthuis, respectively named as Group T, Group W and Group WT were set. Shrimp in Group T were cultured at 18 C without any treatment; Shrimp in Group W and Group WT were cultured at 18 C and then injected with WSSV. At 24 hours post challenge, the water temperature of Group T and Group WT was increased to 25 C. Cephalothorax samples were collected from Group W and Group WT at 12 hours post temperature rising and RNA-seq was performed. Totally 61,886 unigenes with an average length of 626 bp were obtained, among which about 6,421 were differentially expressed genes (DEGs). 4689 DEGs were down-regulated in Group WT and the other 1,732 DEGs were up-regulated, of which 173 were WSSV genes. According to the Gene Ontology analysis, the DEGs were involved in various biological