Production and bioactivity of recombinant tilapia IL-$1\beta$

  • Hong, Su-Hee (Faculty of Marine Bioscience and Technology, Gangneung-Wonju National University)
  • Published : 2009.08.01

Abstract

To study the biological activity of interleukin-$1\beta$(IL-$1\beta$), a proinflammatory cytokine, in nile tilapia, Oreochromis niliticus, the recombinant tilapia IL-$1\beta$ was produced in E. coli cells based on pQE vector. Ni-NTA (nitriloacetic acid) metal affinity chromatography was used to purify recombinant protein. The eluted fractions exhibited a single band of protein with a molecular weight of about 25kDa, which is in close agreement with 25.4 kDa predicted by the cDNA sequence. The biological activity of the purified recombinant tilapia IL-$1\beta$ was tested through its effects on IL-$1\beta$ gene expression, which are known as IL-$1\beta$ inducible genes in mammals and fishes. IL-$1\beta$ gene expression induced by poly I:C, a synthetic double stranded RNA, was also assessed in tilapia head kidney cells. IL-$1\beta$ gene expression was analysed using QPCR (quantitative polymerase chain reaction). The ratio of the indicated gene expression was expressed as the relative mRNA level to $\beta$-actin mRNA level, which is constitutively expressed in macrophages. Consequently, head kidney cells incubated for three hours with rIL-$1\beta$(10, 2, 1 $\mu{g}$/ml) showed a dose dependent increase in IL-$1\beta$ mRNA levels and 1 $\mu{g}$/ml of poly I:C was also able to induce IL-$1\beta$ gene expression in head kidney in tilapia.

References

  1. Alvi, S.A., Rajasingam, D., Brown, N.L., Elder, M.G., Bennett, P.R. and Sullivan, M.H.: Theproduction of IL-1beta from human fetal membranes is not obligatory for increased prostaglandin output. Immunology, 97: 249-256, 1999 https://doi.org/10.1046/j.1365-2567.1999.00769.x
  2. Bird, S., Zou, J. and Secombes, C.J.: Evolution ofIL-1$\beta$gene in vertebrates. EMBL accessionNo. AJ010640, 1998
  3. Blecha, F., Reddy, D.N., Chitko-Mckown, C.G., Mcvey, D.S., Chengappa, M.M., Goodband, R.D. and Nelssen, J.L.: Influence of recombinant bovine interleukin-1$\beta$and interleukin-2 in pigs vaccinated and challenged with Streptococcus suis. Vet. Immunol. & Immunopathol., 44: 329-346, 1995 https://doi.org/10.1016/0165-2427(94)05301-8
  4. Cannon, J.G., Clark, B.D., Wingfield. P., Schmeissner,U., Losberger, C., Dinarello, C.A. and Shaw, A.R.: Rabbit IL-1: Cloning, expression, biological properties, and transcription during endotoxemia. The journal ofimmunology, 142(7): 2299-2306, 1989
  5. Clem, L.W., Sizemore, R.C., Ellsaesser, C.F. and Miller, N.W.: Monocytes as accessory cells in fish immune responses. Developmental and comparative Immunology, 9: 803-809, 1985 https://doi.org/10.1016/0145-305X(85)90046-1
  6. Diaz, A., Chepenik, K.P., Korn, J.H., Reginato, A.M. and Jimenez, S.A.: Differential regulation of cycloxygenases 1 and 2 by interluekin-1 beta, Tumor necrosis factor-alpha, and transforming growth factor-beta 1 in human lung fibroblasts. Experimental cell research, 241: 222-229, 1998 https://doi.org/10.1006/excr.1998.4050
  7. Dinarello, C. .: Interleukin 1. Rev. Infect. Dis., 6:51-95, 1984 https://doi.org/10.1093/clinids/6.1.51
  8. Dinarello, C.A., Ikejima, T., Warner, S.J.C., Orencole, S.F., Lonneman, G., Cannon, J.G. and Libby, P.: Interleukin 1 induces interleukin1. I. Induction of circulating interleukin 1 in rabbits in vivo and in human mononuclear cells in vitro. J. immunology, 139: 1902, 1987
  9. Flodstrom, M., Chen, M.- C., Smismans, A., Schuit, F., Pipeleers, D.G. and Eizirik, D.L.: IL-1beta increases arginin accumulation and activates the citrulline-NO cycle in rat pancreatic $\beta$-cells. Cytokine, 11(6):400-407, 1999 https://doi.org/10.1006/cyto.1998.0446
  10. Fuhlbrigge, R.C., chaplin, D. D., Kielly, J.M. and Unanue, E.R.: Regulation of IL-1 gene expression by adherence and LPS. J. Immunol., 138:3799-3802, 1987
  11. Fujiki, K., Nakao, M., Shin, D. and Yano, T.: cDNAcloning of carp homologue of mammalianIL-1$\beta$. EMBL accession No. AB010701,1998
  12. Haskill, S., Johnson, C., Eierman, D., Becker, S.and Warren, K.: Adherence induces selective mRNA expression of monocyte mediators and proto-oncogenes. J. Immunol.,140:1690-1694, 1988
  13. Holland, J.: The generation of eicosanoids in rainbow trout with particular reference to gills and leukocytes. Ph. D thesis, University of Wales, 1997
  14. Jobling, S.A., Auron, P.E., Gurka, G., Webb, A.C., Mc donald, B., Rosenwasser, L.J. and Gehrke, L.: Biological activity and receptor binding of human prointerleukin-1beta and subpeptids. J. Biol. Chem., 31:16372-16378, 1988
  15. Komaki, G., Arimura, A. and Koves, K.: Effects of intravenous injection of IL-1$\beta$on PGE2 levels in several brain areas as determined by microdialysis. Am. J. Physiol., 262:E246- E251, 1992
  16. Laing, K.J., Grabowski, P.S., Belosevic, M., Secombes, C.J.: A partial sequence for nitric oxide synthase from a goldfish (Carassius auratus) macrophage cell line. Immunol. Cell Biol., 74(4):374-379, 1996 https://doi.org/10.1111/j.1440-1711.1996.tb03930.x
  17. Lee, D.-S., Hong, S., Lee, H.-J., Jun, L.-J., Chung, J.-K., Kim, K.-H. and Jeong, H.-D.: Molecular cDNA cloning and analysis of the organization and expression of the IL-1$\beta$gene in the nile tilapia, Oreochromis niliticus. Com. Bio. Phy. partA, 143:307-314, 2006 https://doi.org/10.1016/j.cbpa.2005.12.014
  18. Schindler, R., Clark, B.D. and Dinarello, C.A.: Dissociation between IL-1$\beta$mRNA and protein synthesis in human peripheral blood mononuclear cells. J. Biol. Chem. 265:10232-10234, 1990
  19. Secombes, C., Zou, J., Daniels, G., Cunningham,C., Koussounadis, A. and Kemp, G.: Rainbowtrout cytokine and cytokine receptorgenes. Immunological reviews, 166: 333-340, 1998 https://doi.org/10.1111/j.1600-065X.1998.tb01273.x
  20. Sigel, M.M., Hamby, B.A. and Huggins, E.M.Jr.: Phylogenetic studies on lymphokines. Fish lymphocytes respond to human IL-1 and epithelial cells produce an IL-1-like factor. Verterinary immunology and immuno pathology, 12:47-58, 1986 https://doi.org/10.1016/0165-2427(86)90109-1
  21. Smith, K.A., Lachman, L.B., Oppenheim, J.J. andFavata, M.M.: The functional relationship ofthe interleukins. J. Exp. Med. 151:1551-1556, 1980 https://doi.org/10.1084/jem.151.6.1551
  22. Verburg Van Kemenade, B.M.L., Weyts, F.A.A., Debets, R. and Flik, G.: Carp macrophages and neutrophil granulocytes secrete an IL-1- like factor. Developmental and comparative Immunology, 19:59-70, 1995 https://doi.org/10.1016/0145-305X(94)00047-J
  23. Warner, S.J.C., Auger, K.R. and Libby, P.: Carp macrophages and neutrophil granulocytes secrete an IL-1- like factor. The journal of immunology, 139(6):1911-1917, 1987
  24. Weyts, F.A.A., Rombout, J.H.W.M., Flik, G. and Verburg Van Kemenade, B.M.L.: A commoncarp leucocyte cell line shares morphologic and functional characteristic withmacrophages. Fish and shell fish immunology,7:123-133, 1997 https://doi.org/10.1006/fsim.1996.0069
  25. Zou, J., Grabowski, P.S., Cunningham, C. and Secombes,C.J.: Molecular cloning of interleukin-1beta from Rainbow trout reveals noevidence of an ice cut site. Cytokine, 11(8):552-560, 1999 https://doi.org/10.1006/cyto.1998.0470
  26. Zou, J., Cunningham, C. and Secombes, C.J.: The Rainbow trout Oncorhynchus mykiss interleukin- 1$\beta$gene has a different organization to mammals and undergoes incomplete splicing. Eur. J. Biochem., 259(3):901-908, 1999 https://doi.org/10.1046/j.1432-1327.1999.00121.x