人類腸胃道中未受控制的腸道免疫便會造成慢性IBD，就像Crohn disease與潰瘍性結腸炎 (UC)。IBD是一群慢性的疾病，它會導致大腸或小腸中的發炎與潰瘍。在之前研究中發現，介白素十重組蛋白與經基因改造細菌所分泌介白素十可以改善由糊精硫酸鈉 (DSS)所引發的老鼠結腸炎。近來，人類介白素十九被界定為介白素十的家族之ㄧ。它會影響T細胞成熟並改變Th1/Th2平衡而偏向Th2。再者，介白素十九是介白素十的轉錄活化因子，且在介白素十九刺激的周邊血單核球細胞中會引發高量的介白素十上升。在本篇研究中，我們利用減毒豬霍亂沙門氏桿菌攜帶表現老鼠介白素十九的質體以減緩DSS所引發的老鼠急性結腸炎，並減緩其致死率與減輕其許多臨床上的發炎指標。我們也發現在介白素十九所治療的結腸炎老鼠中，在早期便可以誘發介白素十的上升而達到預防DSS所引發的老鼠結腸炎。在未來利用減毒豬霍亂沙門氏桿菌攜帶表現老鼠介白素十九的治療將提供ㄧ個新的臨床應用的方法來治療人類IBD.

Uncontrolled mucosal immunity in the gastrointestinal tract of humans results in chronic inflammatory bowel disease (IBD), such as Crohn’s disease and ulcerative colitis. IBD is a group of chronic disorder that causes inflammation or ulceration in the small and/or large intestines. In previous studies, recombinant interleukin (IL)-10 and genetically modified bacteria secreting IL-10 ameliorated the dextran sulfate sodium (DSS)-induced colitis of IBD model. Recently, human IL-19 has been identified as a member of the IL-10 cytokine family. It influences the maturation of T cells and alters the balance of Th1/Th2 cells in favor of Th2. Furthermore, IL-19 is a transcriptional activator of IL-10 and IL-10 is strongly induced in IL-19-stimulated peripheral blood mononuclear cells. In this study, we investigated whether the expression of murine IL-19 gene was beneficial in ameliorating IBD. Our results showed that the use of the attenuated Salmonella choleraesuis carrying murine IL-19 expressing plasmid for IBD gene therapy reduced the mortality and many clinical markers in DSS- induced acute colitis as compared with those untreated counterparts. We also found that IL-10 was induced in IL-19 treated colitis mice and it prevented the progression at the early stage of DSS-induced colitis. The S. choleraesuis encoding IL-19 for the treatment of IBD provides a new strategy in future clinical application.

Boirivant, M., Fuss, I. J., Chu, A., and Strober, W. (1998). Oxazolone colitis: A murine model of T helper cell type 2 colitis treatable with antibodies to interleukin 4. J Exp Med 188, 1929-1939.

Bouma, G., and Strober, W. (2003). The immunological and genetic basis of inflammatory bowel disease. Nat Rev Immunol 3, 521-533.

Buelens, C., Verhasselt, V., De Groote, D., Thielemans, K., Goldman, M., and Willems, F. (1997). Human dendritic cell responses to lipopolysaccharide and CD40 ligation are differentially regulated by interleukin-10. Eur J Immunol 27, 1848-1852.

Elson, C. O. (2000). Commensal bacteria as targets in Crohn's disease. Gastroenterology 119, 254-257.

Fiocchi, C. (1997). Intestinal inflammation: a complex interplay of immune and nonimmune cell interactions. Am J Physiol 273, G769-775.

Fiocchi, C. (1998). Inflammatory bowel disease: etiology and pathogenesis. Gastroenterology 115, 182-205.

Furrie, E., Macfarlane, S., Kennedy, A., Cummings, J. H., Walsh, S. V., O'Neil D, A., and Macfarlane, G. T. (2005). Synbiotic therapy (Bifidobacterium longum/Synergy 1) initiates resolution of inflammation in patients with active ulcerative colitis: a randomised controlled pilot trial. Gut 54, 242-249.

Gallagher, G., Dickensheets, H., Eskdale, J., Izotova, L. S., Mirochnitchenko, O. V., Peat, J. D., Vazquez, N., Pestka, S., Donnelly, R. P., and Kotenko, S. V. (2000). Cloning, expression and initial characterization of interleukin-19 (IL-19), a novel homologue of human interleukin-10 (IL-10). Genes Immun 1, 442-450.

Gallagher, G., Eskdale, J., Jordan, W., Peat, J., Campbell, J., Boniotto, M., Lennon, G. P., Dickensheets, H., and Donnelly, R. P. (2004). Human interleukin-19 and its receptor: a potential role in the induction of Th2 responses. Int Immunopharmacol 4, 615-626.

Groux, H., O'Garra, A., Bigler, M., Rouleau, M., Antonenko, S., de Vries, J. E., and Roncarolo, M. G. (1997). A CD4+ T-cell subset inhibits antigen-specific T-cell responses and prevents colitis. Nature 389, 737-742.

Hans, W., Scholmerich, J., Gross, V., and Falk, W. (2000). Interleukin-12 induced interferon-gamma increases inflammation in acute dextran sulfate sodium induced colitis in mice. Eur Cytokine Netw 11, 67-74.

Ito, R., Shin-Ya, M., Kishida, T., Urano, A., Takada, R., Sakagami, J., Imanishi, J., Kita, M., Ueda, Y., Iwakura, Y., et al. (2006). Interferon-gamma is causatively involved in experimental inflammatory bowel disease in mice. Clin Exp Immunol 146, 330-338.

Jordan, W. J., Eskdale, J., Boniotto, M., Lennon, G. P., Peat, J., Campbell, J. D., and Gallagher, G. (2005). Human IL-19 regulates immunity through auto-induction of IL-19 and production of IL-10. Eur J Immunol 35, 1576-1582.

Kitajima, S., Takuma, S., and Morimoto, M. (1999). Tissue distribution of dextran sulfate sodium (DSS) in the acute phase of murine DSS-induced colitis. J Vet Med Sci 61, 67-70.

Kruis, W., Fric, P., Pokrotnieks, J., Lukas, M., Fixa, B., Kascak, M., Kamm, M. A., Weismueller, J., Beglinger, C., Stolte, M., et al. (2004). Maintaining remission of ulcerative colitis with the probiotic Escherichia coli Nissle 1917 is as effective as with standard mesalazine. Gut 53, 1617-1623.

Kuhn, R., Lohler, J., Rennick, D., Rajewsky, K., and Muller, W. (1993). Interleukin-10-deficient mice develop chronic enterocolitis. Cell 75, 263-274.

Liao, Y. C., Liang, W. G., Chen, F. W., Hsu, J. H., Yang, J. J., and Chang, M. S. (2002). IL-19 induces production of IL-6 and TNF-alpha and results in cell apoptosis through TNF-alpha. J Immunol 169, 4288-4297.

Lindsay, J., Van Montfrans, C., Brennan, F., Van Deventer, S., Drillenburg, P., Hodgson, H., Te Velde, A., and Sol Rodriguez Pena, M. (2002). IL-10 gene therapy prevents TNBS-induced colitis. Gene Ther 9, 1715-1721.

Lindsay, J. O., Ciesielski, C. J., Scheinin, T., Hodgson, H. J., and Brennan, F. M. (2001). The prevention and treatment of murine colitis using gene therapy with adenoviral vectors encoding IL-10. J Immunol 166, 7625-7633.

Lindsay, J. O., Sandison, A., Cohen, P., Brennan, F. M., and Hodgson, H. J. (2004). IL-10 gene therapy is therapeutic for dextran sodium sulfate-induced murine colitis. Dig Dis Sci 49, 1327-1334.

Mannon, P. J., Fuss, I. J., Mayer, L., Elson, C. O., Sandborn, W. J., Present, D., Dolin, B., Goodman, N., Groden, C., Hornung, R. L., et al. (2004). Anti-interleukin-12 antibody for active Crohn's disease. N Engl J Med 351, 2069-2079.

Melgar, S., Karlsson, A., and Michaelsson, E. (2005). Acute colitis induced by dextran sulfate sodium progresses to chronicity in C57BL/6 but not in BALB/c mice: correlation between symptoms and inflammation. Am J Physiol Gastrointest Liver Physiol 288, G1328-1338.

Monteleone, G., Biancone, L., Marasco, R., Morrone, G., Marasco, O., Luzza, F., and Pallone, F. (1997). Interleukin 12 is expressed and actively released by Crohn's disease intestinal lamina propria mononuclear cells. Gastroenterology 112, 1169-1178.

Moore, K. W., de Waal Malefyt, R., Coffman, R. L., and O'Garra, A. (2001). Interleukin-10 and the interleukin-10 receptor. Annu Rev Immunol 19, 683-765.

Nagalakshmi, M. L., Murphy, E., McClanahan, T., and de Waal Malefyt, R. (2004). Expression patterns of IL-10 ligand and receptor gene families provide leads for biological characterization. Int Immunopharmacol 4, 577-592.

Neish, A. S., Gewirtz, A. T., Zeng, H., Young, A. N., Hobert, M. E., Karmali, V., Rao, A. S., and Madara, J. L. (2000). Prokaryotic regulation of epithelial responses by inhibition of IkappaB-alpha ubiquitination. Science 289, 1560-1563.

Neurath, M. F., Finotto, S., and Glimcher, L. H. (2002). The role of Th1/Th2 polarization in mucosal immunity. Nat Med 8, 567-573.

Nielsen, O. H., Kirman, I., Rudiger, N., Hendel, J., and Vainer, B. (2003). Upregulation of interleukin-12 and -17 in active inflammatory bowel disease. Scand J Gastroenterol 38, 180-185.

Okayasu, I., Hatakeyama, S., Yamada, M., Ohkusa, T., Inagaki, Y., and Nakaya, R. (1990). A novel method in the induction of reliable experimental acute and chronic ulcerative colitis in mice. Gastroenterology 98, 694-702.

Omata, F., Birkenbach, M., Matsuzaki, S., Christ, A. D., and Blumberg, R. S. (2001). The expression of IL-12 p40 and its homologue, Epstein-Barr virus-induced gene 3, in inflammatory bowel disease. Inflamm Bowel Dis 7, 215-220.

Osman, N., Adawi, D., Ahrne, S., Jeppsson, B., and Molin, G. (2004). Modulation of the effect of dextran sulfate sodium-induced acute colitis by the administration of different probiotic strains of Lactobacillus and Bifidobacterium. Dig Dis Sci 49, 320-327.

Paglia, P., Terrazzini, N., Schulze, K., Guzman, C. A., and Colombo, M. P. (2000). In vivo correction of genetic defects of monocyte/macrophages using attenuated Salmonella as oral vectors for targeted gene delivery. Gene Ther 7, 1725-1730.

Parronchi, P., Romagnani, P., Annunziato, F., Sampognaro, S., Becchio, A., Giannarini, L., Maggi, E., Pupilli, C., Tonelli, F., and Romagnani, S. (1997). Type 1 T-helper cell predominance and interleukin-12 expression in the gut of patients with Crohn's disease. Am J Pathol 150, 823-832.

Pestka, S., Krause, C. D., Sarkar, D., Walter, M. R., Shi, Y., and Fisher, P. B. (2004). Interleukin-10 and related cytokines and receptors. Annu Rev Immunol 22, 929-979.

Pletnev, S., Magracheva, E., Kozlov, S., Tobin, G., Kotenko, S. V., Wlodawer, A., and Zdanov, A. (2003). Characterization of the recombinant extracellular domains of human interleukin-20 receptors and their complexes with interleukin-19 and interleukin-20. Biochemistry 42, 12617-12624.

Podolsky, D. K. (2002). Inflammatory bowel disease. N Engl J Med 347, 417-429.
Sartor, R. B. (2004). Therapeutic manipulation of the enteric microflora in inflammatory bowel diseases: antibiotics, probiotics, and prebiotics. Gastroenterology 126, 1620-1633.
Schmidt, C., Giese, T., Ludwig, B., Mueller-Molaian, I., Marth, T., Zeuzem, S., Meuer, S. C., and Stallmach, A. (2005). Expression of interleukin-12-related cytokine transcripts in inflammatory bowel disease: elevated interleukin-23p19 and interleukin-27p28 in Crohn's disease but not in ulcerative colitis. Inflamm Bowel Dis 11, 16-23.

Shiau, A. L., Chen, C. C., Yo, Y. T., Chu, C. Y., Wang, S. Y., and Wu, C. L. (2005). Enhancement of humoral and cellular immune responses by an oral Salmonella choleraesuis vaccine expressing porcine prothymosin alpha. Vaccine 23, 5563-5571.

Shiau, A. L., Chen, Y. L., Liao, C. Y., Huang, Y. S., and Wu, C. L. (2001). Prothymosin alpha enhances protective immune responses induced by oral DNA vaccination against pseudorabies delivered by Salmonella choleraesuis. Vaccine 19, 3947-3956.

Steidler, L., Hans, W., Schotte, L., Neirynck, S., Obermeier, F., Falk, W., Fiers, W., and Remaut, E. (2000). Treatment of murine colitis by Lactococcus lactis secreting interleukin-10. Science 289, 1352-1355.

Stevceva, L., Pavli, P., Husband, A. J., and Doe, W. F. (2001). The inflammatory infiltrate in the acute stage of the dextran sulphate sodium induced colitis: B cell response differs depending on the percentage of DSS used to induce it. BMC Clin Pathol 1, 3.

Uhlig, H. H., and Powrie, F. (2003). Dendritic cells and the intestinal bacterial flora: a role for localized mucosal immune responses. J Clin Invest 112, 648-651.

van Deventer, S. J., Elson, C. O., and Fedorak, R. N. (1997). Multiple doses of intravenous interleukin 10 in steroid-refractory Crohn's disease. Crohn's Disease Study Group. Gastroenterology 113, 383-389.

Weiss, S., and Chakraborty, T. (2001). Transfer of eukaryotic expression plasmids to mammalian host cells by bacterial carriers. Curr Opin Biotechnol 12, 467-472.

Yen, D., Cheung, J., Scheerens, H., Poulet, F., McClanahan, T., McKenzie, B., Kleinschek, M. A., Owyang, A., Mattson, J., Blumenschein, W., et al. (2006). IL-23 is essential for T cell-mediated colitis and promotes inflammation via IL-17 and IL-6. J Clin Invest 116, 1310-1316.

Yuhua, L., Kunyuan, G., Hui, C., Yongmei, X., Chaoyang, S., Xun, T., and Daming, R. (2001). Oral cytokine gene therapy against murine tumor using attenuated Salmonella typhimurium. Int J Cancer 94, 438-443.
Zdanov, A. (2004). Structural features of the interleukin-10 family of cytokines. Curr Pharm Des 10, 3873-3884.