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- Title
Cloning and expression of the first nonmammalianinterleukin-11gene in rainbow troutOncorhynchus mykiss.
- Authors
Wang, Tiehui; Holland, Jason W.; Bols, Niels; Secombes, Christopher J.
- Abstract
Interleukin (IL)-11 is a multifunctional cytokine that stimulates hematopoietic progenitor cells and exerts a series of important immunomodulatory effects. It was believed to be restricted to mammals, but here we report the first nonmammalianIL-11gene, in rainbow trout (Oncorhynchus mykiss). A trout IL-11 cDNA clone was isolated that contains a 5′-untranslated region (UTR) of 400 bp, an open reading frame of 612 bp and a large 3′-UTR of 1924 bp. Analysis of a genomic DNA clone from a trout lambda library revealed that the troutIL-11gene has the same five exon/four intron gene organization, as well as the same intron phase, as mammalianIL-11genes. The 204 amino acid trout IL-11 translation has a predicted signal peptide of 26 amino acids and mature peptide of 178 amino acids, with a calculated molecular mass of 20.5 kDa and a theoretical pIof 9.83. The mature peptide contains a cysteine residue and a potential N-linked glycosylation site that are not present in mammals. Phylogenetic analysis clearly grouped trout IL-11 with IL-11 molecules from other species and separated from other members of the IL-6 family. TheIL-11gene is highly expressed in intestine and gills in healthy fish and its expression can also be detected in spleen, head kidney, brain, skin and muscle. Bacterial infection of rainbow trout markedly up-regulatesIL-11expression in liver, head kidney and spleen.IL-11expression is also up-regulated in RTS-11 cells (a trout macrophage cell line), which constitutively expressed the lowest level ofIL-11of the four trout cell lines examined, after stimulation with bacteria, lipopolysaccharide, poly(I:C) and recombinant trout IL-1β. Only a single transcript of 3.2 kb could be detected in lipopolysaccharide or recombinant IL-1β-stimulated RNA samples by northern blotting. The expression results, showing that IL-11 is widely distributed and modulated by infection and other cytokines, suggest that fish IL-11 is an active player in the cytokine network and the host immune response to infection.
- Subjects
INTERLEUKINS; CYTOKINES; CELLS; HEMATOPOIETIC growth factors; RNA; GENES; MAMMALS
- Publication
FEBS Journal, 2005, Vol 272, Issue 5, p1136
- ISSN
1742-464X
- Publication type
Article
- DOI
10.1111/j.1742-4658.2005.04544.x