PUBLICATION
Molecular characterization, expression analysis, and biological effects of interleukin-8 in grass carp Ctenopharyngodon idellus
- Authors
- Wang, T.T., Song, X.H., Bao, G.M., Zhao, L.X., Yu, X., and Zhao, J.
- ID
- ZDB-PUB-140728-31
- Date
- 2013
- Source
- Fish & shellfish immunology 35(5): 1421-1432 (Journal)
- Registered Authors
- Keywords
- Gene expression pattern; Grass carp Ctenopharyngodon idellus; Interleukin-8 (IL-8); Phylogenetic analysis; Proinflammatory activity
- MeSH Terms
-
- Amino Acid Sequence
- Analysis of Variance
- Animals
- Base Sequence
- Carps/genetics*
- Carps/immunology
- Cloning, Molecular
- Cluster Analysis
- Computational Biology
- DNA Primers/genetics
- DNA, Complementary/genetics
- Gene Expression Profiling/veterinary
- Gene Expression Regulation/genetics
- Gene Expression Regulation/immunology*
- Interleukin-8/chemistry
- Interleukin-8/genetics*
- Interleukin-8/metabolism
- Models, Molecular*
- Molecular Sequence Data
- Phylogeny
- Protein Conformation*
- Real-Time Polymerase Chain Reaction/veterinary
- Sequence Alignment
- Sequence Analysis, DNA/veterinary
- PubMed
- 23994423 Full text @ Fish Shellfish Immunol.
Citation
Wang, T.T., Song, X.H., Bao, G.M., Zhao, L.X., Yu, X., and Zhao, J. (2013) Molecular characterization, expression analysis, and biological effects of interleukin-8 in grass carp Ctenopharyngodon idellus. Fish & shellfish immunology. 35(5):1421-1432.
Abstract
Interleukin-8 (IL-8) is a CXC chemokine that plays key regulatory roles in the immune and inflammatory responses implicated in many human diseases. In this study, we identified and characterized an IL-8 homologue from the grass carp, Ctenopharyngodon idellus. A sequence alignment of the full-length cDNA and genomic DNA showed that the exon/intron organization of grass carp IL-8 (gcIL-8) is identical to those of other known CXC chemokine genes. A multiple alignment analysis showed that gcIL-8 is an ELR(-)CXC chemokine, and its deduced amino acid sequence shares 81% and 36% identity with common carp IL-8s L1 (GenBank ID: ABE47600) and L2 (GenBank ID: AB470924), respectively, suggesting that it belongs to the lineage 1 group of fish IL-8 proteins. On a phylogenetic tree, gcIL-8 clustered with other teleost IL-8 proteins to form a fish-specific clade, clearly distinct from those of bird, mammal, and amphibian proteins. Real-time quantitative PCR analysis indicated that gcIL-8 is differentially expressed in various tissues under normal conditions and that the expression of gcIL-8 mRNA in immune-related tissues is clearly upregulated by Aeromonas hydrophila infection. To explore the biological effects of gcIL-8, we produced a recombinant protein, rgcIL-8, in a prokaryotic expression system. Purified rgcIL-8 was confirmed to be chemoattractive for head kidney neutrophils and mononuclear leukocytes in vitro. Our histopathological study also revealed that rgcIL-8 exerts proinflammatory effects by inducing neutrophil infiltration and erythrocyte extravasation. Overall, these results suggest that IL-8 is crucially involved in the inflammatory responses of fish.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping