Earticle

초록

영어

CXCL8 (interleukin-8, IL-8) is a CXC family chemokine that recruits specific target cells and mediates inflammation and wound healing. This study reports the identification and characterization of two cxcl8 homologs from rock bream, Oplegnathus fasciatus. Investigation of molecular signature, homology, phylogeny, and gene structure suggested that they belonged to lineages 1 (L1) and 3 (L3), and designated Ofcxcl8-L1 and Ofcxcl8-L3. While Ofcxcl8-L1 and Ofcxcl8-L3 revealed quadripartite and tripartite organization, in place of the mammalian ELR (Glu-Leu-Arg) motif, their peptides harbored EMH (Glu-Met-His) and NSH (Asn-Ser-His) motifs, respectively. Transcripts of Ofcxcl8s were constitutively detected by Quantitative Real-Time PCR (qPCR) in 11 tissues examined, however, at dierent levels. Ofcxcl8-L1 transcript robustly responded to treatments with stimulants, such as flagellin, concanavalin A, lipopolysaccharide, and poly(I:C), and pathogens, including Edwardsiella tarda, Streptococcus iniae, and rock bream iridovirus, when compared with Ofcxcl8-L3 mRNA. The dierences in the putative promoter features may partly explain the dierential transcriptional modulation of Ofcxcl8s. Purified recombinant OfCXCL8 (rOfCXCL8) proteins were used in in vitro chemotaxis and proliferation assays. Despite the lack of ELR motif, both rOfCXCL8s exhibited leukocyte chemotactic and proliferative functions, where the potency of rOfCXCL8-L1 was robust and significant compared to that of rOfCXCL8-L3. The results, taken together, are indicative of the crucial importance of Ofcxcl8s in inflammatory responses and immunoregulatory roles in rock bream immunity.

목차

Abstract
1. Introduction
2. Materials and Methods
2.1. Chemicals, Reagents, Experimental Fish, and Immune Stimulants
2.2. Animal Rearing and Ethics
2.3. Determination of Complementary- and Genomic-DNA (cDNA and gDNA) Sequences of Ofcxcl8 Homologs
2.4. Molecular Profiling of Rock Bream cxcl8 Homologs by Computational Tools
2.5. Transcriptional Profiling of Rock Bream cxcl8 Homologs
2.6. Functional Characterization of Rock Bream OfCXCL8 Recombinant Proteins
2.7. Statistical Analysis
3. Results
3.1. Molecular Profiling of Structural Features
3.2. Profiling of Genomic Arrangement and Promoter Sequences
3.3. Profiling of Transcriptional Gene Expression of Rock Bream cxcl8 (il-8) Homologs
3.4. Functional Characterization of Rock Bream CXCL8 Homologs Using Recombinant Proteins
4. Discussion
5. Conclusions
References

키워드

저자

• Navaneethaiyer Umasut [ Department of Marine Life Sciences & Fish Vaccine Research Center, Jeju National University, Department of Ocean Sciences, Memorial University of Newfoundland, St. John’s, NL A1C 5S7, Canada ]
• SDNK Bathige [ Department of Marine Life Sciences & Fish Vaccine Research Center, Jeju National University, Sri Lanka Institute of Nanotechnology (SLINTEC), Nanotechnology and Science Park, Mahenwatta, Pitipana, Homagama 10206, Sri Lanka ]
• William Shanthakumar Thulasitha [ Department of Marine Life Sciences & Fish Vaccine Research Center, Jeju National University, Department of Zoology, Faculty of Science, University of Jaffna 4000, Sri Lanka ]
• Minyoung Oh [ Department of Marine Life Sciences & Fish Vaccine Research Center, Jeju National University, Department of Biochemistry and Molecular Biology, Dalhousie University ]
• Jehee Lee [ Department of Marine Life Sciences & Fish Vaccine Research Center, Jeju National University, Marine Science Institute, Jeju National University, Jeju Self-Governing Province ] Corresponding Author

참고문헌

발행기관

간행물

표지보기 관심저널 등록

• 간행물명

  해양과학연구소 연구논문집 [Bulletin of the Marine Science Institute]

• 간기

  폐간

• pISSN

  1225-5734

• 수록기간

  1977~2021

• 십진분류

  KDC 454 DDC 551