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Hwang, Jee Youn,Jeong, Ji-Min,Kwon, Mun-Gyeong,Seo, Jung Soo,Hwang, Seong Don,Son, Maeng-Hyun,Bae, Jin-Sol,Choi, Kwang-Min,Cho, Dong-Hee,Jeswin, Joseph,Park, Chan-Il Elsevier 2018 FISH AND SHELLFISH IMMUNOLOGY Vol.73 No.-
<P>Coinhibitory pathways in the B7-CD28 family provide critical inhibitory signals that regulate immune homeostasis, defense and protect tissue integrity. CD276 (B7-H3) is an important immune checkpoint member of this family, which is induced on antigen-presenting cells (APCs), and plays an important role in the inhibition of T cell function. We have characterized the CD276 gene of olive flounder, Paralichthys olivaceus. OfCD276 has an ORF of 912 bp that codes for 303 amino acids with a predicted molecular mass of 33 kDa. It is a type I trans membrane protein with a single extracellular V- and C-like Ig domains, a transmembrane region, and a highly diverse cytoplasmic tail. This gene was distinctly expressed in gill, spleen, and skin, and sparsely expressed in other tissues. Pathogen stimulation by VHSV revealed that transcription of OfCD276 was induced on early hours in liver and expressed late in head kidney, spleen, intestine and gill tissues. Flow cytometry analysis of leukocytes revealed the percentage of granulocytes and lymphocytes that expressed OfCD276 molecules on their cell surface was 85.1% and 3.1%, respectively. Our study shows a significant role played by this coinhibitory molecule that participate in the regulation of the cell mediated immune response.</P>
Hwang, Jee Youn,Ahn, Sang Jung,Kwon, Mun-Gyeong,Seo, Jung Soo,Hwang, Seong Don,Son, Maeng-Hyun Elsevier 2017 FISH AND SHELLFISH IMMUNOLOGY Vol.66 No.-
<P><B>Abstract</B></P> <P>Interferon-inducible protein 56 (<I>IFI56,</I> also known as ISG56/<I>IFIT1</I>, interferon-induced protein with tetratricopeptide repeats 1) is strongly induced in response to interferon and a potent inhibitor of viral replication and translational initiation. Here, we describe the identification of <I>IFI56</I> (<I>OfIFI56</I>) in olive flounder, its characteristic features, and expression levels in various tissues before and after viral hemorrhagic septicemia virus (VHSV) infection. The full-length <I>OfIFI56</I> sequence was identified from rapid amplification of cDNA ends PCR. The complete coding sequence of <I>OfIFI56</I> is 1971 bp in length and encodes 431 amino acids. The putative <I>OfIFI56</I> protein has multiple tetratricopeptide (TPR) motifs, which regulate diverse biological processes, such as organelle targeting, protein import, and vesicle fusion. Based on sequence analysis, the <I>Larimichthys crocea</I> IFI56 protein (61%) had the highest sequence homology to <I>OfIFI56.</I> In healthy olive flounder, <I>OfIFI56</I> mRNA expression was detected in many tissues such as intestine, gill, head kidney, heart, spleen, and trunk kidney tissues. After VHSV challenge, <I>OfIFI56</I> mRNA was significantly up-regulated in these tissues. Additionally, <I>OfIFI56</I> expression was induced by poly I:C but not by <I>Streptococcus parauberis</I> and <I>S. iniae</I> infection or lipopolysaccharide injection in kidney and spleen tissues of olive flounder. These results demonstrate that piscine <I>OfIFI56</I> expression is not induced by bacterial infection but is selectively induced by viral infection, especially VHSV, and that <I>OfIFI56</I> may play an important role in the host response against VHSV infection.</P> <P><B>Highlights</B></P> <P> <UL> <LI> The fish IFI56/IFIT1 family including OfIFI56 grouped with IFI56 and not fish IFI58. </LI> <LI> OfIFI56 mRNA was up-regulated in all tissues except muscle after VHS virus challenge. </LI> <LI> OfIFI56 expression was not induced by <I>Streptococcus parauberis</I> and <I>S. iniae</I> infection and LPS injection in olive flounder. </LI> <LI> OfIFI56 mRNA expression markedly increased in all tissues after poly I: C infection. </LI> </UL> </P>
Hwang, Jee Youn,Markkandan, Kesavan,Kwon, Mun Gyeong,Seo, Jung Soo,Yoo, Seung-il,Hwang, Seong Don,Son, Maeng-Hyun,Park, Junhyung Elsevier 2018 FISH AND SHELLFISH IMMUNOLOGY Vol.76 No.-
<P><B>Abstract</B></P> <P>Olive flounder (<I>Paralichthys olivaceus</I>) is one of the most valuable marine aquatic species in South Korea and faces tremendous exposure to the viral hemorrhagic septicemia virus (VHSV). Given the growing importance of flounder, it is therefore essential to understand the host defense of <I>P. olivaceus</I> against VHSV infection, but studies on its immune mechanism are hindered by the lack of genomic resources. In this study, the <I>P. olivaceus</I> was infected with disease-causing VHSV isolates, ADC-VHS2012-11 and ADC-VHS2014-5 which showed moderate virulent (20% mortality) and high virulent (65% mortality), in order to investigate the effect of difference in pathogenicity in head kidney during 1, 3, 7 days of post-infection using Illumina sequencing. After removing low-quality sequences, we obtained 144,933,160 high quality reads from thirty-six libraries which were further assembled into 53,384 unigenes with an average length of 563 bp with a range of 200 to 9605 bp. Transcriptome annotation revealed that 30,475 unigenes with a cut-off e-value of 10<SUP>−5</SUP> were functionally annotated. In total, 10,046 unigenes were clustered into 26 functional categories by searching against the eggNOG database, and 22,233 unigenes to 52 GO terms. In addition, 12,985 unigenes were grouped into 387 KEGG pathways. Among the 13,270 differently expressed genes, 6578 and 6692 were differentially expressed only in moderate and high virulent, respectively. Based on our sequence analysis, many candidate genes with fundamental roles in innate immune system including, pattern recognition receptors (TLRs & RLRs), Mx, complement proteins, lectins, and cytokines (chemokines, IFN, IRF, IL, TRF) were differentially expressed. Furthermore, GO enrichment analysis for these genes revealed gene response to defense response to virus, apoptotic process and transcription factor activity. In summary, this study identifies several putative immune pathways and candidate genes deserving further investigation in the context of novel gene discovery, gene expression and regulation studies and lays the foundation for fish immunology especially in <I>P. olivaceus</I> against VHSV.</P> <P><B>Highlights</B></P> <P> <UL> <LI> First RNA-seq based transcriptome of moderate and high virulent VHSV-infected <I>P. olivaceus</I> head kidney. </LI> <LI> The assembly contig length is more compared to the other olive flounder transcriptome published so far. </LI> <LI> Genes involved in innate immune response were showed significantly differentially upregulated during day 1 of high virulent and day 3 of moderate virulent. </LI> <LI> This <I>de novo</I> transcriptome provides useful information for evaluating the molecular mechanisms underlying host-virus interaction. </LI> </UL> </P>
Hwang, Jee Youn,Lee, Seongdo,Priyathilaka, Thanthrige Thiunuwan,Yang, Hyerim,Kwon, Hyukjae,Kwon, Mun Gyeong,Hwang, Seong Don,Kim, Myoung-Jin,Lee, Jehee Elsevier 2018 Aquaculture Vol.484 No.-
<P><B>Abstract</B></P> <P>Viral hemorrhagic septicemia is a serious disease that can assume epidemic proportions in cultivated olive flounders in Korea. The causal agent of this disease is viral hemorrhagic septicemia virus (VHSV), and it leads to vast economic losses in the olive flounder aquaculture industry. Therefore, the rapid and accurate detection of VHSV is paramount. At present, the manual of the World Organization for Animal Health's Office International des Epizooties (OIE) for pathogen detection is being followed for VHSV detection in Korea. However, in that manual, the primers for VHSV detection are based on the European VHSV genotype Ia. In this study, we identified 5 VHSV strains from olive flounders in Korea, and the nucleotide and amino acid sequences of VHSV genes (N, P, M, and G) were compared to investigate the genetic variation of VHSV genotypes. As expected, VHSV isolates from Korea were highly related to genotype IVa, clearly differing from the 3 European genotypes. In addition, the N gene showed low genetic variation and therefore might be considered a useful marker for VHSV detection, regardless of genotype. In contrast, the other VHSV genes (P, M, and G) had different nucleotides within genotypes. They might be suitable for designing specific primers for distinguishing between various VHSV genotypes. Duplex RT-PCR using the newly designed primers successfully detected all VHSV isolates and validated the genotype (I and IV) without sequence analysis. According to the results, all the VHSV isolates were successfully detected, and their genotypes were validated by duplex RT-PCR using the newly designed primers. Collectively, our findings suggest that duplex PCR is a convenient and appropriate method for the diagnosis of VHSV isolates in Korean aquaculture systems.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Nucleotide and amino acid sequence variation of VHSV genes (N, P, M, and G) were identified. </LI> <LI> A newly developed duplex-PCR simultaneously detect VHSV and distinguish the genotypes of VHSV without sequence analysis. </LI> <LI> Evaluation of newly designed primers using Korean VHSV isolates. </LI> </UL> </P>
Hwang, Jee Youn,Markkandan, Kesavan,Han, Kyudong,Kwon, Mun Gyeong,Seo, Jung Soo,Yoo, Seung-il,Hwang, Seong Don,Ji, Bo Young,Son, Maeng-Hyun,Park, Jun-hyung Springer-Verlag 2018 Genes & Genomics Vol.40 No.3
<P>Olive flounder (Paralichthys olivaceus) is one of the most economically important aquaculture fish. However, its production is often affected by various diseases, especially viral hemorrhagic septicemia virus (VHSV) that cause serious economic losses. In this study, we sequenced the whole transcriptome of the P. olivaceus using Illumina RNA-sEq. De novo assembly of control and virus-infected cDNA libraries of head kidney at 13 and 20 A degrees C was accomplished with 2,007,532,438 raw reads, resulting in 244,578 unigenes with an average length of 533 bp and found 65,535 candidate coding unigenes with homology to other species by BLAST analysis. DEG analysis among control and virus-infected head kidney samples of 13 and 20 A degrees C revealed that 1290 up-regulated and 162 down-regulated genes (p ae<currency> 0.01), linked to metabolism, virulence factors, adhesion and immune-response. We constructed an expressed gene catalog for the P. olivaceus to serve as a resource for marine environmental genomic and immuno-genetic/genomic studies focused on uncovering the molecular mechanisms underlying the responses of P. olivaceus to VHSV under different temperature.</P>