Lamprey (Lethenteron japonicum) IL-17 upregulated by LPS-stimulation in the skin cells
- 260 Downloads
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.
KeywordsCyclostomes Lamprey Skin Cytokine IL-17
This work was supported by JSPS Grant-in Aid for Young Scientists (Start Up) 18880025.
- Asahida T, Kobayashi T, Saitoh K, Nakayama I (1996) Tissue preservation and total DNA extraction from fish stored at ambient temperature using buffers containing high concentration of urea. Fish Sci 62:727–730Google Scholar
- Endo Y, Nonaka M, Saiga H, Kakinuma Y, Matsushita A, Takahashi M, Matsushita M, Fujita T (2003) Origin of mannose-binding lectin-associated serine protease (MASP)-1 and MASP-3 involved in the lectin complement pathway traced back to the invertebrate, amphioxus. J immunol 170:4701–4707PubMedGoogle Scholar
- Fossiez F, Djossou O, Chomarat P, Flores-Romo L, Ait-Yahia S, Maat C, Pin JJ, Garrone P, Garcia E, Saeland S, Blanchard D, Gaillard C, Das Mahapatra B, Rouvier E, Golstein P, Banchereau J, Leubecque S (1996) T cell interleukin-17 induces stromal cells to produce proinflammatory and hematopoietic cytokines. J Exp Med 183:2593–603PubMedCrossRefGoogle Scholar
- Gonzalez SF, Huising MO, Stakauskas R, Forlenza M, Verburg-van Kemenade BML, Buchmann K, Nielsen E, Wiegertjes GF (2006b) Real-time gene expression analysis in carp (Cyprinus carpio L.) skin: Inflammatory responses to injury mimicking infection with ectoparasites. Dev Comp Immunol 31:244–254PubMedCrossRefGoogle Scholar
- Good RA, Finstad J, Litman GW (1972) Immunology. In: Hardisty MV, Potter IC (eds) The biology of lampreys, Vol. 2. Academic, London, pp 405–432Google Scholar
- Hymowitz SG, Filvaroff EH, Yin JP, Lee J, Cai L, Risser P, Maruoka M, Mao W, Foster J, Kelley RF, Pan G, Gurney AL, de Vos AM, Starovasnik MA (2001) IL-17s adopt a cystine knot fold: structure and activity of a novel cytokine, IL-17F, and implications for receptor binding. EMBO J 20:5332–5341PubMedCrossRefGoogle Scholar
- Potter IC, Percy LR, Barber DL, Macey DJ (1972) The morphology, development and physiology of blood cells. In: Hardisty MV, Potter IC (eds) The biology of lampreys, Vol. 2. Academic, London, pp 233–289Google Scholar
- Shi Y, Ullrich SJ, Zhang J, Connolly K, Grzegorzewski KJ, Barber MC, Wang W, Wathen K, Hodge V, Fisher CL, Olsen H, Ruben SM, Knyazev I, Cho YH, Kao V, Wilkinson KA, Carrell JA, Ebner R (2000) A novel cytokine receptor-ligand pair. Identification, molecular characterization, and in vivo immunomodulatory activity. J Biol Chem 275:19167–19176PubMedCrossRefGoogle Scholar
- Terajima D, Yamada S, Uchino R, Ikawa S, Ikeda M, Shida K, Arai Y, Wang HG, Satoh N, Satake M (2003) Identification and sequence of seventy-nine new transcripts expressed in hemocytes of Ciona intestinalis, three of which may be involved in characteristic cell-cell communication. DNA Res 10:203–212PubMedCrossRefGoogle Scholar
- Whitear M (1984) Epidermis. In: Bereiter-Hahn J, Matoltsy AG, Richards KS (eds) Biology of the integument, 2 Vertebrates. Springer-Verlag, Tokyo, pp 8–38Google Scholar