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- 저자 : Jee-Youn Hwang (검색결과 158 건)
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Hwang, Jee Youn,Kwon, Mun Gyeoung,Jung, Sung-Hee,Park, Myoung Ae,Kim, Dong-Wook,Cho, Wang Sik,Park, Jeong Woo,Son, Maeng-Hyun Elsevier 2017 FISH AND SHELLFISH IMMUNOLOGY Vol.62 No.-
<P><B>Abstract</B></P> <P>Viral hemorrhagic septicemia (VHS) is a highly contagious disease of cultured flounder caused by VHS virus (VHSV). To develop effective VHSV vaccines, it is essential to understand the molecular mechanisms underlying the host's protective response against VHSV. The purpose of this study is to clarify which genes are involved in the protective response of olive flounder after VHSV vaccination. We first injected olive flounder intraperitoneally with 10<SUP>7</SUP> TCID<SUB>50</SUB> heat-inactivated VHSV vaccine and evaluated the vaccine efficacy at 20 °C. Fish vaccinated with heat-inactivated VHSV were significantly protected compared to non-vaccinated fish, with a relative percentage survival of 83%. To analyze the vaccination-induced changes in the expression profiles of genes, kidneys were collected from control and vaccinated fish at days 1, 3, and 7 after vaccination and global gene expression profiling was carried out by RNA sequencing. The analysis revealed that 15,001 genes were differentially expressed by at least 2-fold between vaccinated fish and non-vaccinated controls. Of these, 58 genes clustered into the acute phase response, Toll-like receptor, interferon-inducible/regulatory proteins, and apoptosis pathways. These data provided insights into the molecular mechanisms underlying the protective immune response of olive flounder against heat-inactivated VHSV vaccine and might aid future studies to develop a highly immunogenic vaccine against VHSV in flounder.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Viral hemorrhagic septicemia virus (VHSV) is a major pathogen of cultured flounder. </LI> <LI> We analyzed the transcriptomic response to heat-inactivated VHSV in olive flounder. </LI> <LI> In total, 15,001 genes were differentially expressed after vaccination. </LI> <LI> Data analysis delineated 4 functional pathways responsive to VHSV vaccination. </LI> <LI> These findings can aid the future development of highly immunogenic VHSV vaccines. </LI> </UL> </P>
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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>
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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>
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Hwang, Jee Youn,Kwon, Mun-Gyeong,Kim, Yu Jin,Jung, Sung-Hee,Park, Myoung-Ae,Son, Maeng-Hyun Elsevier 2017 Fish & shellfish immunology Vol.60 No.-
<P><B>Abstract</B></P> <P>Vaccination by immersion is suitable for mass vaccination of small size fish. However, no viral vaccine has been developed for immersion applications, because of low efficacy. In this study, we evaluated the efficacy and safety of immersion vaccine against viral hemorrhagic septicemia (VHS) containing Montanide IMS 1312 VG adjuvant in olive flounder (<I>Paralichthys olivaceus</I>). Healthy fish were vaccinated by an immersion method with a heat-inactivated FP-VHS2010-1 strain of VHS virus (VHSV) in combination with Montanide IMS 1312 VG for 5 min at 20 ± 2 °C. The control group was vaccinated with sterile PBS. No toxicity of immersion vaccine with Montanide IMS 1312 VG adjuvant was observed by hematological and histopathological analysis. Immersion vaccine with adjuvant enhanced gene expression of immune-associated genes, i.e., genes encoding interleukin (IL)-1β, IL-6, IL-8, and Toll-like receptor (TLR) 3. Relative percent survival (RPS) of fish was measured on weeks 4 and 8 post vaccination. In fish vaccinated with adjuvant, RPS was significantly higher than that of fish vaccinated without adjuvant. The results of the present study provide evidence that the VHSV immersion vaccine with Montanide IMS 1312 VG induces protective immunity in olive flounder against VHS.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Immersion vaccination of heat-inactivated VHSV with Montanide 1312 VG protected olive flounder from VHS. </LI> <LI> Immersion vaccination inceased the levels of several immune genes. </LI> <LI> Addition of Montanide 1312 VG further increased the levels of immune genes. </LI> <LI> Duration of protective effect of immersion vaccination was short. </LI> </UL> </P>
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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>
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NV gene variations of viral hemorrhagic septicemia virus decrease cellular energy metabolism
Hwang, Jee Youn,Cho, Mi Young,Hwang, Seong Don,Park, Myoung Ae,Cho, Hyun Kook,Kim, So Young,Sung, Mi So,Kim, Dong Uk,Cheong, JaeHun Elsevier Scientific Pub. Co 2016 Aquaculture Vol.464 No.-
<P><B>Abstract</B></P> <P>The viral hemorrhagic septicemia virus (VHSV) is a viral pathogen to cause severe losses in farming industry of olive flounder. Among six-encoding VHSV proteins, NV protein has been shown to have an impact on virulence. Thirteen VHSV strains are isolated from olive flounders showing VHS symptoms and mortality in Korea, and then molecular variation of nucleotide and amino acid sequences of the NV genes were compared. Although common mutations were detected at four sites in nucleotide sequences of NV gene, there was no difference in the amino acid sequences among 13 VHSV variants. We identified substitution of one base of 34 positions within the NV ORF of 13 VHSV isolates and these point mutations resulted in changes in amino acid sequences at 6 residues of NV (8, 56, 81, 88, 117, and 119). Based on finding of NV-mediated decrease of intracellular ATP level, protein expression of six NV variants in flounder cells decreased more than control NV protein. The strongest decreasing activity on cellular ATP level was shown by No. 56 (serine→leucine) of NV variation. The variation changes the hydropathy index significantly. The results imply that we can use appearance of some amino acid variations as virulence markers in VHSV-derived mortality.</P> <P><B>Statement of relevance</B></P> <P>1. Recently VHSV has infected >80 kinds of fish species. The number of infection host of VHSV is rapidly increased. Among the VHSV-infected fishes, olive flounder is one of the most important fish of aquaculture in Korea, Japan, and China. In every year, we found the massive VHSV-derived mortality of olive flounder aquaculture in Korea. To overcome this VHSV pathogenesis, we need to identify high-pathogenic VHSV appearance at aquaculture in earlier time.</P> <P>2. Since VHSV gene is highly mutated in time-dependent manner, the efficient vaccine for VHSV-derived disease prevention is not well achieved. Therefore, the other approaches for prevention of epidemics by VHSV should be required. Our research goal is to develop prevention of epidemics propagation of VHSV infection by high pathogenic VHSV prediction with gene variation analysis.</P> <P>3. In this study, we identified several gene variations of VHSV NV protein from 13 VHSV specimens of severe VHS-showing olive flounders in Korea. The gene variations are associated with amino acid changes of VHSV NV proteins. The amino acid variants of NV protein show the inhibitory activity of fish cellular energy (ATP) generation. The cellular energy deprivation might give a big impact on fish health and mortality. The DNA mutation on specific amino acid changes is possibly applied to develop the PCR diagnosis system for early detection of high pathogenic VHSV appearance. The early pathogenic gene diagnosis of VHSV gives more efficient prevention of epidemics.</P> <P><B>Highlights</B></P> <P> <UL> <LI> We have identified nucleotide and amino acid variations of VHSV NV gene. </LI> <LI> VHSV NV gene expression in olive flounder cells induces cellular energy deprivation. </LI> <LI> Four different amino acid variants of VHSV NV protein show differential inhibition of cellular ATP generation intensity. </LI> <LI> The most decreased cellular ATP level is shown by No. 56 (S -> L) variant, showing the most dramatic change of the hydropathy index. </LI> </UL> </P>
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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>
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