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研究生:張淑菁
研究生(外文):Shu-Ching Chang
論文名稱:利用LAE技術製造抗IL-13抗體作為診斷試劑
論文名稱(外文):Generation of anti-IL13 antibodies for diagnostic agent by LAE technique
指導教授:黃昭蓮陳浩仁
學位類別:碩士
校院名稱:國立中正大學
系所名稱:分子生物研究所
學門:生命科學學門
學類:生物科技學類
論文種類:學術論文
論文出版年:2005
畢業學年度:93
語文別:中文
論文頁數:47
中文關鍵詞:介白素13線性重複抗原決定區
外文關鍵詞:IL-13、LAE
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介白素13 (IL-13) 是由第二型輔助性T淋巴球分泌的一種細胞素,參與調節一些免疫反應,例如在氣喘、過敏、皮膚性過敏及發炎症狀裡血液中的IL-13濃度會增加。此外,IL-13可促進B細胞分化,使B細胞產生IgE,進而調節一些與發炎前期相關的細胞激素產生,並可增加MHCⅡ (major histocompatibility complex Ⅱ) 分子抗原呈現。這些臨床觀察意味著血液中IL-13含量可以作為一些免疫疾病診斷之指標。本研究期望製造抗IL-13之抗體,以作為開發快速檢測細胞激素之抗體晶片的材料。根據結構模型,選擇位於IL-13分子表面之不同區段作為抗原決定區 (epitopes),利用線性排列重複抗原技術,製造含線性重複抗原決定區之多胜肽 (linear array epitope),並與綠膿桿菌外毒素A接受器結合區(PEDIa)或麩胱甘肽轉移酵素(GST)結合成融合蛋白。以此純化之融合蛋白作為免疫原,免疫實驗兎與小鼠以刺激體液性免疫反應,並在ELISA檢測發現血清中確實存在抗IL-13不同抗原辨識區之抗體。同時為了製造具高度效價及穩定性之抗體,也利用融合瘤技術,選擇合適效價之小鼠脾臟以製造抗IL-13之單株抗體。目前,已成弗N四段抗原決定區,運用線性排列重複抗原技術得到的免疫原進行兔子免疫後,得到抗IL-13的多株抗體。在所得的抗血清中,發現以IL13-A10、IL13-D7、IL13-E9為抗原所產生之抗體,其分辨原態的IL-13能力皆可達0.5 ng以上。 而在單株抗體製備方面,已完成抗PE-A10與抗PE-E9之融合瘤單株化,並冷凍保存。未來,會進行抗體的純化,將抗體作原態IL-13辨認之分析,並且製備多量抗體以作為細胞激素抗體晶片的材料來源。
Interleukin-13 (IL-13) is a cytokine produced by T helper-2 lymphocytes and plays an important role in regulating several immune responses. The serum concentration of IL-13 increased in asthma, allergy, atopy and inflammatory conditions. It can promote B cell proliferation, induce B cells to produce IgE, down-regulate the production of proinflamatory cytokines and enhance the expression of class II MHC antigens. All these clinical observations implied that the serum concentration of IL-13 could be used as a diagnostic indicator for immunological disorders. We proposed to produce the anti-IL13 antibody, which could detect sub-concentration of IL-13 and be utilized for rapid diagnosis. Base on structural modeling, several different regions that exposed at the molecular surface were selected as antigenic targets. The linear array epitope (LAE) technique was employed to generate DNA fragments encoding for LAE antigen in fusion with Pseudomonas exotoxin domain Ia (PEIa) or glutathione S-transferase (GST). The purified fusion proteins were used to immunize rabbits. Antibodies against IL-13 epitopes were proved to be specific from ELISA assays. So far, we already have generated antigen of IL-13 epitopes by LAE technique, and taken the antigen to immunize rabbit to get the polyclonal antibodies. In these antisera, we found that antibodies against IL13-A10, IL13-D7, and IL13-E9 recognized the minimum quantity of native form IL-13 was 0.5 ng. Furthermore, in order to generate antibodies with high titer and stability, we prepared to generate monoclonal antibodies against IL-13 with hybridoma technique, and accomplished the monoclonal antibody against PE-A10 and PE-E9. The hybridoma cell lines were kept by freezing. In the future, we’ll purify antibodies of ascites to be the material of the antiboby chip.
Andrew, N. J. M. (2000) Regulation of T helper type 2 cell immunity by interleukin-4 and interleukin-13. Pharmacology & Therapeutics 88, 143-151.

Aman, M. J., Tayebi, N., Obiri, N., Puri, R., Modi, W., and Leonard, W. (1996). cDNA cloning and characterisation of the human interleukin-13 receptor ?chain. J Biol Chem 271, 29265-29270.

Barner, M., Mohrs, M., Brombacher, F., and Kopf, M. (1998). Differences between IL-4Ra-deficient and IL-4-deficient mice reveal a role for IL- 13 in the regulation of Th2 responses. Curr Biol 8, 669-672.

Burd, P. R., Thompson, W. C., Max, E. E., and Mills, F. C. (1995). Activated mast cells produce interleukin-13. J Exp Med 181, 1373-1380.

Bernard, J., Treton, D., Vernot-Desroches, C., Boden, C., Horellou, P., Angevin, E., Galanaud, P., Wijdenes, J., and Richard, Y. ( 2001) Expression of interleukin 13 receptor in glioma and renal cell carcinoma. IL13Ra2 as a decoy receptor for IL13. Lab Invesl 81, 1223-1231.

Collighan, N., Giannoudis, P. V., Kourgeraki, O., Perry, S. L., Guillou, P. J., and Bellamy, M. C. (2004) Interleukin 13 and inflammatory markers in human sepsis. British Journal of Surgery 91, 762-768

Chiaramonte, M. G.., Schopf, L. R., Neben, T. Y., Cheever, A. W., Donaldson, D. D., and Wynn, T. A. (1999). IL-13 is a key regulatory cytokine for Th2 cell-mediated pulmonary granuloma formation and IgE responses induced by Schistosoma mansoni eggs. J Immunol 162, 920-930.

Cocks, B. G. (1993) IL13 induces proliferation and differentiation of human B cells activated by the CD40 ligand International Immunology 5, 657-63.
.
Doherty, M., Kastelein, K., Menon, S., Andrade, S., and Coffman, R. (1993). Modulation of murine macrophage function by IL-13. J Immunol 151, 7151-7160.

Debinski, W., Obiri, N. I., Powers, S. K., Pastan, I., and Puri, R.K. (1995) Human glioma cells overexpress receptors for interleukin 13 and are extremely sensitive to a novel chimeric protein composed of interleukin 13 and pseudomonas exotoxin. Clin Cancer Res 1, 1253-1258.
de Waal Malefyt, R., Figdor, C., Huijbens, R., Mohan-Peterson, S., Bennett, B., Culpepper, J., Dang, W., Zurawski, G., & de Vries, J. (1993). Effects of IL-13 on phenotype, cytokine production and cytotoxic function of human monocytes. J Immunol 151, 6370-6381.

de Vries, J. E. (1998) The role of IL-13 and its receptor in allergy and inflammatory responses. J Allergy Clin Immunol 102, 165-9.

de Wildt, R. M., Mundy, C. R., Gorick, B.D., and Tomlinson, I. M. (2000) Antibody arrays for high-throughput screening of antibody-antigen interactions. Nat Biotechnol 18, 989-94.

Debinski, W., Obiri, N. I., Pastan, I., and Puri, R. K. (1995) A novel chimeric protein composed of interleukin 13 and Pseudomonas exotoxin is highly cytotoxic to human carcinoma cells expressing receptors for interleukin 13 and interleukin 4. J Biol Chem 270, 16775-16780.

Donaldson, D. D., Whitters, M. J., Fitz, L. J., Neben, T. Y., Finnerty, H., Henderson, S. L., O'Hara, R. M. J., Beier, D. R., Turner, K. J., Wood, C. R., and Collins, M. (1998). The murine IL-13 receptor alpha 2: molecular cloning, characterization, and comparison with murine IL-13 receptor alpha 1. J Immunol 161, 2317-2324.

Emson, C. L., Bell, S. E., Jones, A., Wisden, W., and McKenzie, A. N. J. (1998). Interleukin (IL)-4-independent induction of immunoglobulin (Ig)E, and perturbation of T-cell development in transgenic mice expressing IL-13. J Exp Med 188, 399-404.

Fallon, P. G., Richardson, E. J., McKenzie, G. J., and McKenzie, A. N. J. (2000). Schistosome infection of transgenic mice defines distinct and contrasting roles for IL-4 and IL-13: IL-13 is a profibrotic agent. J Immunol 164, 2585-2591.

Finkelman, F., Shea-Donohue, T., Goldhill, J., Sullivan, C., Morris, S., Madden, K., Gause,W., and Urban, J. (1997). Cytokine regulation of host defence against parasitic gastrointestinal nematodes: lessons from studies with rodent models. Annu Rev Immunol 15, 505-533.

Frazer, K. A., Ueda, Y., Zhu, Y., Gifford, V. R., Garofalo, M. R., Mohandas, N., Martin, C. H., Palazzolo, M. J., Cheng, J. F., & Rubin, E. M. (1997). Computational and Biol analysis of 680 kb of DNA sequence from the human 5q31 cytokine gene cluster. Genome Res 7, 495-512.

Franklin, J. M., Elizabeth, D., James, W., and Robert, P. (2001) Solution structure of human IL-13 and inflammatory markers in human sepsis. J. Mol. Biol 310, 219-230.
Grunig, G., Warnock, M., Wakil, A. E., Venkayya, R., Brombacher, F., Rennick, D. M., Sheppard, D., Mohrs, M., Donaldson, D. D., Locksley, R. M., and Corry, D. B. (1998). Requirement for IL-13 independently of IL-4 in Exp asthma. Science 282, 2261- 2263.

Holmgren, J., Lycke, N., and Czerkinsky, C. (1993) Cholera toxin and cholera B subunit as oral-mucosal adjuvant and antigen vector systems. Vaccine 11, 1179–1184.

Hilton, D. J., Zhang, J. G., Metcalf, D., Alexander, W., Nicola, N., and Wilson, T. (1996). Cloning and characterisation of a binding subunit of the interleukin-13 receptor that is also a component of the interleukin-4 receptor. Proc Natl Acad Sci USA 93, 497-501.

Hsu, C. T., Ting, C. Y., Ting, C. J., Chen, T. Y., Lin, C. P., Jacqueline, W. P., and
Hwang, J. (2000) Vaccination against Gonadotropin-releasing Hormone (GnRH) Using Toxin Receptor-binding Domain-conjugated GnRH Repeats. Cancer Res 60, 3701-3705.

Husain, S. R., Joshi, B. H., and Puri, R.K. (2001) Interleukin-13 receptor as a unique target for anti-glioblastoma therapy. Int J Cancer 92, 168-175.

Husain, S. R., and Puri, R. K.(2003) Interleukin-13 receptor-directed cytotoxin for malignant glioma therapy: from bench to bedside. J Neurooncol 65, 37-48.

He, X.Y., Verma, N., Plain, K., and Hall, B. M. (1999) Interleukin 13 Cloning From DA Rats. Transplantation Proceedings 31, 1572–1573.

Keegan, A., Johnston, J., Tortolani, P., McReynolds, L., Kinzer, C., O'Shea, J., and Paul, W. (1995). Similarities and differences in signal transduction by interleukin-4 and interleukin-13: analysis of Janus kinase activation. Proc Natl Acad Sci USA 92, 7681-7685.

Kawakami, M., Leland, P., and Kawakami, K. (2001) Mutation and functional analysis of IL-13 receptors in human malignant glioma cells. Oncol Res 12, 459-467.

Kawakami, K., Kioi, M., Liu, Q., Kawakami, M., and Puri, R. K. (2005) Evidence that IL-13R alpha2 chain in human glioma cells is responsible for the antitumor activity mediated by receptor-directed cytotoxin therapy. J Immunother 28, 193-202.

Joshi, B. H., Plautz, G. E., and Puri, R.K. (2000) Interleukin-13 receptor alpha chain: a novel tumor-associated transmembrane protein in primary explants of human malignant gliomas. Cancer Res 60, 1168-72.

Lai, Y. H., and Mosmann, T. R. (1999). Mouse IL-13 enhances antibody production in vivo and acts directly on B-cells in vitro to increase survival and hence antibody production. J Immunol 162, 78-87.

Liu, H., Jacobs, B. S., Liu, J., Prayson, R. A., Estes, M. L., Barnett, G.H., and Barna, B. P. (2000) Interleukin-13 sensitivity and receptor phenotypes of human glial cell lines: non-neoplastic glia and low-grade astrocytoma differ from malignant glioma. Cancer Immunol Immunother 49, 319-324.

Lu, F. M., Chou, C. C., Chiang, B. L., and Hsieh, K. H., (1998) Immunologic changes during immunotherapy in asthmatic children: increased IL-13 and allergen-specific IgG4 antibody levels. Ann Allergy Asthma Immunol 80, 419-23.

Mintz, A., Gibo, D. M., Slagle, W. B., Christensen, N.D., and Debinski, W. (2002) IL-13Ra2 is a glioma-restricted receptor for interleukin-13. Neoplasia 4, 388-399.

Mitomu, K., Koji, k., and Raj, K. P. (2004) Mechanism of action of interleukin-13 antagonist (IL-13E13K) in cells expressing various types of IL-4R. Cellular Immunology 229, 41-51.

Matthews, D. J., Emson, C. L., McKenzie, G. J., Jolin, H. E., Blackwell, J. M., and McKenzie, A. N. (2000). IL-13 is a susceptibility factor for leishmania major infection. J Immunol 164, 1458-1462.

McKenzie, G., Bancroft, A., Grencis, R., & McKenzie, A. (1998a). A McKenzie, G.,

McKenzie, A., Culpepper, J., de Waal Malefyt, R., Briere, F., Punnonen, J., Aversa, G., Sato, A., Dang, W., Cocks, B., Menon, S., de Vries, J., Banchereau, J., and Zurawski, G. (1993a). Interleukin-13, a T-cell-derived cytokine that regulates human monocyte and B-cell function. Proc Natl Acad Sci USA 90, 3735-3739.

McKenzie, A., Li, X., Largaespada, D., Sato, A., Atsushi, K., Zurawski, S.,Doyle, E., Milatovich, A., Francke, U., Copeland, N., Jenkins, N., and Zurawski, G. (1993b). Structural comparison and chromosomal locationof the human and mouse IL-13 genes. J Immunol 150, 5436-5444.

Minty, A., Chalon, P., and Derocq, J. M. (1993) Interleukin-13 is a new human lymphokine regulating inflammatory an immune responses. Nature 362, 248-250.

Puri, R. K., Leland, P., Obiri, N. I., Husain, S. R., Kreitman, R. J., Haas, G. P., Pastan, I., and Debinski, W. (1996) Targeting of interleukin-13 receptor on human renal cell carcinoma cells by a recombinant chimeric protein comprised of interleukin 13 and pseudomonas exotoxin. Blood 87, 4333-4349.

Sylvie, L., Natalio, V., Renee, R., Daniel, C., and Pascual, F. (1995) IL-13 and IL-4 share signal transduction elements as well as receptor components in TF-1 cells. FEBS Letter 366, 122-126.

Wills, K. M., Luyimbazi, J., Xu, X., Schofield, B., Neben, T. Y., Karp, C. L., and Donaldson, D. D. (1998). Interleukin-13: central mediator of allergic asthma. Science 282, 2258-2261.

Wills, K. M., and Chiaramonte, M. (2003) Interleukin-13 in asthma. Curr Opin Pulm Med 9, 21-7.

Yang, G.., Volk, A., Petley, T., Emmell, E., Giles-Komar, J., Shang, X., Li, Jian., Anuk, M., Das., Dave S., Don, E., Griswold., and Li Li. (2004) Anti-IL-13 monoclonal antibody inhibits airway hyperresponsiveness, inflammation and airway remodeling. Cytokine 28, 224-32.

Zurawski, G., and de Vries, J. E. (1994) Interleukin 13, an interleukin 4-like cytokine that acts on monocytes and B cells, but not on T cells. Immunology Today 15,19-26.
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