An EST-based approach for identifying genes expressed in the gills of olive flounder Paralichthys olivaceus

이정호,노재구,김현철,박철지,민병화,김영옥,김종현,김경길,명정인,김우진 한국어병학회 2009 한국어병학회지 Vol.22 No.3

Analysis of expressed sequence tags (ESTs) is an efficient approach for gene discovery, expression profiling, and development of resources useful for functional genomics studies. As part of studies on the immune system of olive flounder, a total of 251 EST sequences from gill cDNA library were generated to identify and characterize important genes in the immune machanisms of olive flounder. Of the 251 clones, 126 clones (50.2%) were identified as orthologues of known genes from olive flounder and other organisms. Among the 126 EST clones, 16 clones (12.7%) were representing 9 unique genes identified as homologous to the previously reported olive flounder ESTs, 100 clones (79.4%) representing 103unique genes were identified as orthologs of known genes from other organisms. We also identified several kinds of immune associated proteins, indicating EST as a powerful method for identifying immune related genes of fish as well as identifying novel genes. Further studies using cDNA microarrays are needed to identify the differentially expressed transcripts after disease infection. Analysis of expressed sequence tags (ESTs) is an efficient approach for gene discovery, expression profiling, and development of resources useful for functional genomics studies. As part of studies on the immune system of olive flounder, a total of 251 EST sequences from gill cDNA library were generated to identify and characterize important genes in the immune machanisms of olive flounder. Of the 251 clones, 126 clones (50.2%) were identified as orthologues of known genes from olive flounder and other organisms. Among the 126 EST clones, 16 clones (12.7%) were representing 9 unique genes identified as homologous to the previously reported olive flounder ESTs, 100 clones (79.4%) representing 103unique genes were identified as orthologs of known genes from other organisms. We also identified several kinds of immune associated proteins, indicating EST as a powerful method for identifying immune related genes of fish as well as identifying novel genes. Further studies using cDNA microarrays are needed to identify the differentially expressed transcripts after disease infection.

Immune gene mining by pyrosequencing in the rockshell, <i>Thais clavigera</i>

Rhee, Jae-Sung,Kim, Bo-Mi,Jeong, Chang-Bum,Horiguchi, Toshihiro,Lee, Young-Mi,Kim, Il-Chan,Lee, Jae-Seong Elsevier 2012 FISH AND SHELLFISH IMMUNOLOGY Vol.32 No.5

<P><B>Abstract</B></P><P>The rockshell, <I>Thais clavigera</I> (Gastropoda: Muricidae) has been shown to be a useful species as a potential indicator for diverse pollution in the marine environment. However, their genetic information is still not widely available. Here, we performed an extensive transcriptome analysis of <I>T. clavigera</I> using the pyrosequencing method, and selected innate immune-related genes. Among the unigenes obtained in this species, we annotated a number of immune system-related genes (e.g. adhesive protein, antimicrobial protein, apoptosis- and cell cycle-related protein, cellular defense effector, immune regulator, pattern recognition protein, protease, protease inhibitor, reduction/oxidation-related protein, signal transduction-related protein and stress protein), which are potentially useful for immunity research in this species. To confirm the usefulness of potential immune-biomarker genes, we checked the transcript level of specific immune genes in both different tissues and LPS-exposed rockshells within the <I>T. clavigera</I> transcript database. This study would be helpful to extend our knowledge on the immune system of rockshell in comparative aspects. Also it would be useful to develop the rockshell as a potential test organism for monitoring of marine environment quality.</P> <P><B>Highlights</B></P><P>► Novel investigation of extensive immune-relevant genes in the rockshell, <I>Thais clavigera</I> as the first report. ► Confirm the usefulness for comparative analysis of specific immune gene in different developmental tissues of rockshell. ► Development of potential immune-biomarker gene upon bacterial challenge. ► Contribution on immune-relevant gene information resource for comparative gene expression analysis within aquatic invertebrates.</P>

Identification of a novel immune-related gene in the immunized black soldier fly, Hermetia illucens (L.)

( Seong-tae Jung ),( Tae-won Goo ),( Seong Ryul Kim ),( Gwang-ho Choi ),( Sung-wan Kim ),( Pham Thi Nga ),( Seung-won Park ) 한국잠사학회 2018 International Journal of Industrial Entomology Vol.36 No.2

The larvae of Hermetia. illucens have a high probability of coming into contact with microorganisms such as bacteria and fungi. Therefore, the survival of H. illucens is primarily the protection of their own against microbial infection. This effect depends on the development of the innate immune system. Antimicrobial Peptides (AMPs) exhibit antimicrobial activity against other bacterial strains and can provide important data to understand the basis of the innate immunity of H. illucens. In this study, we injected larvae with Enterococcus. faecalis (gram-positive bacteria) and Serratia. marcescens as (gram-negative bacteria) to test the hypothesis that H. illucens is protected from infection by its immune-related gene expression repertoire. To identify the inducible immune-related genes, we performed and cataloged the transcriptomes by RNA-Seq analysis. We compared the transcriptomes of whole larvae and obtained a DNA fragment of 465 bp including the poly (A) tail by RACE as a novel H. illucens immune-related gene against bacteria. A novel target mRNA expression was higher in immunized larvae with E. faecalis and S. marcescens groups than non-immunized group. We expect our study to provide evidence that the global RNA-Seq approach allowed for the identification of a gene of interest which was further analyzed by quantitative RT-PCR, together with genes chosen from the available literature.

Identification of a novel immune-related gene in the immunized black soldier fly, Hermetia illucens (L.)

Jung, Seong-Tae,Goo, Tae-Won,Kim, Seong Ryul,Choi, Gwang-Ho,Kim, Sung-Wan,Nga, Pham Thi,Park, Seung-Won Korean Society of Sericultural Science 2018 International Journal of Industrial Entomology Vol.36 No.2

The larvae of Hermetia. illucens have a high probability of coming into contact with microorganisms such as bacteria and fungi. Therefore, the survival of H. illucens is primarily the protection of their own against microbial infection. This effect depends on the development of the innate immune system. Antimicrobial Peptides (AMPs) exhibit antimicrobial activity against other bacterial strains and can provide important data to understand the basis of the innate immunity of H. illucens. In this study, we injected larvae with Enterococcus. faecalis (gram-positive bacteria) and Serratia. marcescens as (gram-negative bacteria) to test the hypothesis that H. illucens is protected from infection by its immune-related gene expression repertoire. To identify the inducible immune-related genes, we performed and cataloged the transcriptomes by RNA-Seq analysis. We compared the transcriptomes of whole larvae and obtained a DNA fragment of 465 bp including the poly (A) tail by RACE as a novel H. illucens immune-related gene against bacteria. A novel target mRNA expression was higher in immunized larvae with E. faecalis and S. marcescens groups than non-immunized group. We expect our study to provide evidence that the global RNA-Seq approach allowed for the identification of a gene of interest which was further analyzed by quantitative RT-PCR, together with genes chosen from the available literature.

Sequencing and comparison of head louse whole genome sequences with those of body louse genome

Jae Soon Kang,Young-Jun Cho,Ju Hyeon Kim,Si Hyeock Lee 한국응용곤충학회 2011 한국응용곤충학회 학술대회논문집 Vol.2011 No.05

The differences in the immune response between body lice, Pediculus humanus humanus, and head lice, Pediculus humanus capitis, were regarded as primary factors determining their differential vector competence. To find any differences in genetic components in immune system between body and head lice, whole genome sequences of head lice were determined by both SBS [sequencing by synthesis, Illumina Genome Analyzer (Illumina-GA)] and pyrosequencing (Roche GS FLX), and compared with the reference genome sequences of body lice. The short DNA reads from Illumina-GA (an average mapping depth of 50-fold) were aligned first to the body louse reference genome, to which Roche GS FLX DNA reads (an average depth of 2.5-fold) were subsequently assembled to make up gaps between mapped consensus. Total consensus showed a size of 114 Mb and a coverage of 96% of the published body louse genome sequences. From this head louse genome sequences, a total of 12,651 genes were predicted and used for comparing with the 10,775 genes previously reported from the body louse genome. The homolog analysis identified 873 head louse-specific genes and 422 body lice-specific genes. Comparison of immune response genes between both louse species showed head lice have more number of immune-related genes than body lice. Head lice were determined to possess all of the 107 immune-related genes reported in the previous study (Kim et al., 2011), suggesting that there is no difference in genetic make-up in terms of the 107 immune-related genes between body and head lice.

유산균 생산 후 폐기되는 부산물 첨가 사료의 급이가 잉어(Cyprinus carpio)의 성장률, 선천성 면역, 면역연관 유전자 발현 및 항균효과에 미치는 영향

최재혁,정상목,양은총,장태원,이찬흔,박관하,최상훈,Choi, Jae Hyeok,Jung, Sang Mok,Yang, Eun Chong,Jang, Tae Won,Lee, Chan Heun,Park, Kwan Ha,Choi, Sanghoon 한국어병학회 2022 한국어병학회지 Vol.35 No.1

This study has been performed to investigate the potential effects of by-product discarded after probiotics production (BPPP) on growth performance, immune gene expression, innate-immunity status, and disease resistance of carp, Cyprinus carpio. For 3 weeks, carp were fed four diets containing different levels of BPPP at 0, 0.1, 0.2 and 0.5% per kg of normal diet. Every 7 days of feeding, immune-related gene expression, serum lysozyme activity and ACH₅₀ were analyzed. Growth rates and challenge test with E. tarda were conducted after 3 weeks of BPPP feeding. Both lysozyme activity and ACH₅₀ were significantly (p<0.05) increased in all BPPP supplemented groups compared to the control at every 7 day for 3 weeks of feeding trial. The gene expression of pro-inflammatory cytokines, IL-1β and TNF-α was significantly (p<0.05) up-regulated until 21 days of feeding in all groups except for 0.2% group on day 7 post feeding. The anti-inflammatory cytokine IL-10 gene expression was only significantly (p<0.05) increased in 0.1% group on day 7 and decreased (p<0.05) on day 14 in all BPPP supplemented groups. On day 21, the IL-10 gene expression was augmented (p<0.05) in all groups. SOD gene expression was significantly (p<0.05) increased compared to the control on day 14 and 21 post feeding, whereas no significant difference was observed on day 7. In challenging test, 0.2%, 0.1%, 0.5% and control group showed 80%, 70%, 60% and 40% of survival rate, respectively. Feed conversion rate was only improved in 0.5% group. In conclusion, the present study indicates that dietary BPPP suplementation improved growth performance, innate immune response and bactericidal activity in carp.

Role of Bacillus subtilis VSG4-derived biosurfactant in mediating immune responses in Labeo rohita

Giri, S.S.,Sen, S.S.,Jun, J.W.,Sukumaran, V.,Park, S.C. Academic Press 2016 FISH AND SHELLFISH IMMUNOLOGY Vol.54 No.-

<P>This study aimed to isolate biosurfactant from CO2-sequestering Bacillus subtilis VSG4 and to evaluate its immunostimulatory effect in Labeo rohita fingerlings. Fish were injected intraperitoneally (i.p.) with 0.1 mL of phosphate-buffered saline (PBS) containing the water-soluble fraction of purified biosurfactant at 50 (S50), 100 (S100), 200 (S200), or 300 (S300) mu g mL(-1). Fish injected with PBS served as controls. Various immunological parameters, including immune-related gene expression, were measured at 14, 21, and 28 days post administration (dpa). At 28 dpa, the.fish were challenged with Aeromonas hydrophila and mortality was recorded up to 14 days. Among the immune parameters tested, lysozyme levels (36.32 +/- 1.79 U mL(-1)), alternative complement pathway activity (76.26 +/- 2.18 U mL(-1)), phagocytic activity (32.18 +/- 0.67%), and serum bactericidal activity (73.2 +/- 4.7%) were significantly higher (P < 0.05) in the 5200 group at 21 dpa than in the controls. Respiratory burst activity (0.386 +/- 0.008 OD630nm) was the highest in the 5200 group at 28 dpa. Of the immune-related genes examined, pro-inflammatory cytokines (TNF-alpha and IL-1 beta) were significantly down-regulated in the S200 and 5300 groups. Expression of anti-inflammatory cytokines (IL-10 and TGF-beta) as well as IKB-alpha was higher (P < 0.05) in the S100-S300 groups at 21 dpa. The expression of NF-kappa B p65, IKK-beta, MAPKp38, and Myd88 was down-regulated in the treated groups when compared to the controls. Fish in the 5200 group exhibited the highest post challenge relative survival rate (67.88%). Collectively, these results suggest that secondary metabolite (biosurfactant) isolated from B. subtilis VSG4 at 200 mu g mL(-1) can positively influence immune responses, enhance disease resistance, and stimulate immune-related gene expression in L rohita. (C) 2016 Elsevier Ltd. All rights reserved.</P>

Calcitonin gene-related peptide is a promising candidate for immune privilege maintenance and restoration via the suppression of ectopic MHC class I

( Long Quan Pi ),( Xing Hai Jin ),( Han Nah Hong ),( Sung Yul Lee ),( Yoon Hee Lee ),( Sungjoo Tommy Hwang ),( Won Soo Lee ) 대한피부과학회 2011 초록집 Vol.49 No.20

e-Poster : Effects of calcitonin gene-related peptide on the immune privilege of human hair follicles (초)

( Long Quan Pi ),( Youn Duk Kim ),( Xing Hai Jin ),( Yoon Hee Lee ),( Jae Hong Ji ),( Sung Yul Lee ),( Han Nah Hong ),( SungJooTommy Hwang ),( Won Soo Lee ) 대한피부과학회 2010 초록집 Vol.48 No.20

Suppressive Effect of Clozapine through Regulation of Chemokine (C-C motif) Ligand Genes in SH-SY5Y Human Neuroblastoma Cells

Hae Jeong Park,Ah Rang Cho,Jin Kyung Park,Won Sub Kang,Jong Woo Kim 대한스트레스학회 2010 스트레스硏究 Vol.18 No.3

비정형 항정신병 치료제인 clozapine은 정신분열병 치료에 매우 효과적인 약물이다. 그러나 clozapine으로 치료 받는 환자의 0.5∼2%에서 무과립구증과 같은 면역계 관련 부작용이 발생한다고 보고되어 왔으며, 따라서 clozapine의 사용은 제한되어 왔다. 본 연구는 microarray 분석법을 이용하여 사람 SH-SY5Y neuroblastoma 세포에서 clozapine (10 μM, 24 h)에 의해 변화되는 유전자의 발현 양상을 알아보고자 하였다. 실험 결과, clozapine에 의해 165개의 유전자 발현이 증가하였으며, 306개의 유전자 발현이 감소하였다. 특히, 면역 관련 유전자의 발현 변화가 현저했으며, chemokine(C-C motif) ligand 3 (CCL3, 15.596-fold)가 가장 크게 증가하였고, retinoic acid early transcript 1E (RAET1E, 0.171-fold)가 가장 크게 감소하는 것을 확인하였다. Clozapine은 특히 CCL3 (MIP-1α), CCL2 (MCP-1) 및 CCR3과 같은 chemokine 유전자의 발현을 증가시켰다. 이러한 결과는 clozapine의 면역계 관련 부작용에 대한 기전을 이해하는데 중요한 자료를 제시할 것으로 생각한다. Clozapine, an atypical antipsychotic agent, has proven to be effective in the treatment of refractory schizophrenia. As the idiosyncratic clozapine-induced adverse effects such as dyslipidemia and agranulocytosis occur in 0.5∼2% of the treated patients, the use of clozapine has been limited. In this study, we assessed the patterns of gene regulation by clozapine (10 μM, 24 h) in SH-SY5Y human neuroblastoma cells through microarray analysis. Clozapine upregulated the expressions of 165 genes, and downregulated the expressions of 305 genes. Of these genes, clozapine potently increased the level of chemokine (C-C motif) ligand 3 gene (CCL3, 15.596-fold) and decreased the level of retinoic acid early transcript 1E (RAET1E, 0.171-fold). Both of these genes belong to the category of immune response-related genes. We also found the changes of the expressions of immune response-related genes were most remarkable. Especially, clozapine increased the levels of chemokine genes, such as CCL3 (also known as MIP-1α), CCL2 (also known as MCP-1), and chemokine (C-C motif) receptor 3 (CCR3). This result may contribute to understanding of adverse effects of clozapine. (Korean J Str Res 2010;18:229∼235)