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標題: 小鼠IL-18s重組蛋白可以刺激小鼠脾臟細胞及EL-4細胞表現IFN-γ
Recombinant Mouse IL-18s can Stimulate Mouse Splenocytes and EL-4 Cells to Express IFN-γ
作者: 林庭而
Lin, Tin-Er
關鍵字: IL-18s;小鼠介白素-18s;IFN-γ
出版社: 獸醫微生物學研究所
引用: Akira, S. (2000). The role of IL-18 in innate immunity. Current opinion in immunology 12, 59-63. Bailey, S.L., Gossner, A., Dalziel, R., and Hopkins, J. (2000). Cloning of Ovine interleukin 18 cDNA. Veterinary Pathology. Conti, B., Jahng, J.W., Tinti, C., Son, J.H., and Joh, T.H. (1997). Induction of interferon-gamma inducing factor in the adrenal cortex. The Journal of biological chemistry 272, 2035-2037. Fournout, S., Dozois, C.M., Yerle, M., Pinton, P., Fairbrother, J.M., Oswald, E., and Oswald, I.P. (2000). Cloning, chromosomal location, and tissue expression of the gene for pig interleukin-18. Immunogenetics 51, 358-365. Gracie, J.A., Robertson, S.E., and McInnes, I.B. (2003). Interleukin-18. Journal of leukocyte biology 73, 213-224. Irmler, I.M., Gajda, M., and Brauer, R. (2007). Exacerbation of antigen-induced arthritis in IFN-gamma-deficient mice as a result of unrestricted IL-17 response. J Immunol 179, 6228-6236. Nakanishi, K., Yoshimoto, T., Tsutsui, H., and Okamura, H. (2001). Interleukin-18 regulates both Th1 and Th2 responses. Annual review of immunology 19, 423-474. Okamura, H., Nagata, K., Komatsu, T., Tanimoto, T., Nukata, Y., Tanabe, F., Akita, K., Torigoe, K., Okura, T., Fukuda, S., et al. (1995a). A novel costimulatory factor for gamma interferon induction found in the livers of mice causes endotoxic shock. Infection and immunity 63, 3966-3972. Okamura, H., Tsutsi, H., Komatsu, T., Yutsudo, M., Hakura, A., Tanimoto, T., Torigoe, K., Okura, T., Nukada, Y., Hattori, K., et al. (1995b). Cloning of a new cytokine that induces IFN-gamma production by T cells. Nature 378, 88-91. Strausberg, R.L., Feingold, E.A., Grouse, L.H., Derge, J.G., Klausner, R.D., Collins, F.S., Wagner, L., Shenmen, C.M., Schuler, G.D., Altschul, S.F., et al. (2002). Generation and initial analysis of more than 15,000 full-length human and mouse cDNA sequences. Proceedings of the National Academy of Sciences of the United States of America 99, 16899-16903. Venkatachalam, K., Mummidi, S., Valente, A.J., Friehs, I., del Nido, P.J., and Chandrasekar, B. (2008). Induction of IL-18 and upregulation of its receptor in a rabbit model of pressure-overload hypertrophy. Medicine. Yang, Y.J., Wang, Z.Y., Chen, S.H., and Ge, X.R. (2005). Cloning and characterization of a new isoform of mouse interleukin-18. Acta biochimica et biophysica Sinica 37, 826-834. Zou, J., Bird, S., Truckle, J., Bols, N., Horne, M., and Secombes, C. (2004). Identification and expression analysis of an IL-18 homologue and its alternatively spliced form in rainbow trout (Oncorhynchus mykiss). European journal of biochemistry / FEBS 271, 1913-1923. 劉雅琳 (2006). 小鼠介白素-2、3、6與18重組蛋白之表現與應用. 碩士論文 中興大學獸醫微生物學研究所。台灣省台中市。中華民國。.
The purposes of this study were to confirm that mouse interleukin-18s (small or short form of interleukin-18, IL-18s) possesses the activity in stimulating mouse immune cells to express IFN-γ gene, to compare the expression of IL-18 and IL-18s mRNAs in mouse organs, and to explore whether other mammals (in particular, human, sheep, and rabbit) also have similar IL-18 isoform.
Interleukin-18 (IL-18) is a multifunctional cytokine. In addition to stimulating the production of IFN-γ, IL-18 can also enhance cytotoxic activity of NK cells, and promote the differentiation of T cells. In recent ten years, researchers have identified a smaller isoform of IL-18 (IL-18s), which is resulted from alternatively spliced IL-18 mRNAs, in rainbow trout, rats, and mice.
In 2005, we and Yang et al. respectively cloned mouse IL-18s and expressed recombinant mouse IL-18s (rmIL-18s) protein. Yang et al. reported that rmIL-18s, unlike IL-18, could not stimulate mouse splenocytes to express IFN-γ. However, in our previous study, rmIL-18s expressed in our laboratory could stimulate mouse splenocytes to express
IFN-γ. In this study, we used rmIL-18 and rmIL-18s expressed in our laboratory to stimulate mouse splenocytes, respectively, and detected the expression of IFN-γ mRNA by reverse transcription-polymerase chain reaction (RT-PCR). Our results confirmed that rmIL-18s could induce mouse splenocytes to express IFN-γ with a comparable activity as recombinant mouse IL-18 (rmIL-18). Moreover, the rmIL-18s could stimulate EL-4 cells (derived from mouse T lymphoma) to express IFN-γ as well.
In addition, we detected and compared the expression of IL-18 and IL-18s mRNAs in mouse organs by RT-PCR. IL-18 mRNA was detected unanimously in all organs examined. However, the distribution of IL-18s mRNA in muse organs was variable among mice. In general, all mice expressed IL-18s mRNA in the muscle and gastrointestinal tract.
Furthermore, we extracted RNA from peripheral blood mononuclear cells (PBMCs) of human, sheep, and rabbit and explored the presence of IL-18s mRNA. We did not observe any alternatively spliced IL-18 mRNA in human and sheep. Interestingly, two alternatively spliced mRNAs of IL-18, which were not in-frame, were identified in rabbit. We expect that the two alternatively spliced IL-18 mRNAs can not produce functional proteins.
其他識別: U0005-2707200916152300
Appears in Collections:微生物暨公共衛生學研究所

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