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Cloning and expression analysis of three striped trumpeter (Latris lineata) pro-inflammatory cytokines, TNF -á and IL-â, in response to infection by the ectoparasitic, Chondracanthus goldsmidi
journal contributionposted on 2023-05-17, 01:04 authored by Covello, JM, Bird, S, Morrison, RN, Battaglene, SC, Seacombes, CJ, Barbara Nowak
This study reports the cloning and sequencing of three striped trumpeter (Latris lineata Forster) proinflammatory cytokines, TNF-a, IL-1b and IL-8, as well as their differential expression in response to an infection by the ectoparasite Chondracanthus goldsmidi. The striped trumpeter TNF-a transcript consisted of 1093 bp, including a 759 bp ORF which translated into a 253 aa transmembrane peptide. The sequence contained a TACE cut site, that would produce a 167 aa soluble peptide containing the TNF ligand family signature. The IL-1b sequence consisted of 963 bp, including a 774 bp ORF which translated into a 258 aa protein. The protein lacked both a signal peptide and an ICE cleavage site, but did contain the IL-1 family signature. The sequence for the chemokine IL-8 contained 906 bp, with an ORF of 297 bp, which translated into a 99 aa protein. The protein lacked an ELR motif as is common with many teleost IL-8 sequences. The differential expression of the three cytokine genes in parasitized fish was investigated via quantitative real-time PCR. A significant up-regulation of all three pro-inflammatory cytokines was found in the gills, which were the site of parasite attachment. Examination of head kidney cells revealed a significant up-regulation of TNF-a, but not IL-1b or IL-8. Conversely, the spleen cells showed significant up-regulation of both IL-1b and IL-8, but not TNF-a. These findings allow for more detailed investigations of the striped trumpeter immune response.
Publication titleFish and Shellfish Immunology
Department/SchoolInstitute for Marine and Antarctic Studies
Place of publicationNetherlands
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