Molecular characterization of a CXCL8-like protein from ayu and its effect on chemotaxis of neutrophils and monocytes/macrophages |
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Authors: | Chang-Qing Chu Xin-Jiang LuChang-Hong Li Jiong Chen |
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Affiliation: | Laboratory of Biochemistry and Molecular Biology, School of Marine Sciences, Ningbo University, Ningbo 315211, China |
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Abstract: | CXCL8, a CXC-type chemokine, plays a crucial role in acute inflammation by recruiting and mediating neutrophils and other cells. In this study, the cDNA and genomic DNA sequence of a CXCL8-like protein (PaCXCL8l) from ayu (Plecoglossus altivelis) was determined. Sequence analysis showed that PaCXCL8l represented the typical structure of animal CXCL8s. Phylogenetic tree analysis indicated that PaCXCL8l was closest to CXCL8 of Atlantic cod (Gadus morhua). Constitutive expression of PaCXCL8l was detected in all tested tissues and monocytes/macrophages, and its expression dramatically increased upon Listonella anguillarum infection. In vitro, recombinant PaCXCL8l exhibited a significant chemotactic effect on neutrophils at 0.1 μg/ml and on monocytes/macrophages at 1.0 μg/ml. In vivo, the numbers of peritoneal neutrophils and monocytes/macrophages were both up-regulated following intraperitoneal administration of recombinant PaCXCL8l. These results suggest that PaCXCL8l is crucially involved in the immune response of ayu by mediating chemotaxis of neutrophils and monocytes/macrophages. |
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Keywords: | aa, amino acids bp, base pairs CFU, colony-forming units CXCL8, CXC Chemokine Ligand 8 hpi, hours post-infection ip, intraperitoneally kDa, kilodaltons MOI, multiplicity of infection NTA, nitrilotriacetic acid ORF, open reading frame RT-qPCR, real-time quantitative PCR SDS&ndash PAGE, sodium dodecyl sulfate polyacrylamide gel electrophoresis |
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