Lamprey (Lethenteron japonicum) IL-17 upregulated by LPS-stimulation in the skin cells.
We report here the first evidence for interleukin-17, a pro-inflammatory cytokine, in cyclostomes. To detect the novel molecules involved in the immune response in the skin of the lamprey Lethenteron japonicum, subtractive hybridization was performed with 6-h-cultured skin cells with or without lipopolysaccharide (LPS). In approximately 100 partially sequenced clones analyzed, we identified an interesting sequence similar to that of the IL-17 genes in teleosts and mammals. Subsequent rapid amplification of cDNA ends was used to obtain the cDNA of lamprey IL-17 (LampIL-17) that contains a 519-bp open reading frame encoding a mature protein of 154 amino acids and a 19-residue NH2-terminal signal peptide. The phylogenetic tree indicated that LampIL-17 is clustered into IL-17D, which is a subgroup of the IL-17 family. Southern blot analysis showed that the lamprey harbors a single copy of the LampIL-17 gene in its genome. The LampIL-17 gene was constitutively expressed in most tissues examined as well as in the skin, where the basal layer epithelial cells expressed LampIL-17 mRNA. Real-time-polymerase chain reaction (RT-PCR) demonstrated that the LampIL-17 gene expression in LPS-stimulated skin cells tended to be greater than that in non-stimulated cells. These results suggest that LampIL-17 is responsible for defense against bacterial infections in the lamprey skin.
Tsutsui
Shigeyuki
S
School of Fisheries Science, Kitasato University, Sanriku, Ofunato, Iwate 022-0101, Japan. tsutsui@kitasato-u.ac.jp
Nakamura
Osamu
O
Watanabe
Tasuku
T
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D017931:DNA Primers / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics; D018628:Gene Dosage; D066298:In Vitro Techniques; D020381:Interleukin-17 / Q000235:genetics*; D007798:Lampreys / Q000033:anatomy & histology / Q000235:genetics* / Q000276:immunology*; D008070:Lipopolysaccharides / Q000494:pharmacology; D008969:Molecular Sequence Data; D010802:Phylogeny; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D012867:Skin / Q000166:cytology / Q000187:drug effects / Q000276:immunology*; D015854:Up-Regulation / Q000187:drug effects
D017931:DNA Primers; D018076:DNA, Complementary; D020381:Interleukin-17; D008070:Lipopolysaccharides; D012333:RNA, Messenger