Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/94758
標題: 比較分析三種異種異體腫瘤抗原之DNA在黑色素瘤小鼠上引起之抗腫瘤免疫反應
作者: 陳柏仰
關鍵字: 黑色素瘤
抗腫瘤免疫反應
異種異體腫瘤抗原
melanoma
xenogenic tumor antigens
DNA-based vaccines
immunotherapy
出版社: 獸醫學系暨研究所
摘要: 黑色素瘤所表現的分化抗原已被證實能夠引起抗腫瘤的細胞毒殺性T細胞及抗體免疫反應。本實驗室及徐維莉老師實驗室架構出三種異種異體多抗原結合位的DNA免疫治療劑:第一種是同時帶有gp100 (280V)、gp100 (210M)及tyrosinase (370D)之異種異體多抗原結合位質體DNA (ME);第二種除了上述的抗原結合位外,再加上Melan-A的質體DNA (MA);最後一種是第二種質體DNA的抗原結合位外,再加上二重複MAGE片段的質體DNA (MAGE)。本實驗的目的是比較這三種以DNA為基礎的免疫療法對B16-F10黑色素瘤小鼠身上引起之免疫反應。首先以螢光顯微鏡及西方墨點法來確認此合成之質體可於真核細胞中表現出抗原及螢光蛋白。而在小鼠試驗中可發現,免疫治療組(ME,MA和MAGE組)具有明顯抑制腫瘤生長及轉移的效果(P < 0.05)。此外西方墨點法分析顯示,所有接受免疫治療的小鼠都可引起相對應的抗原特異性抗體反應。而酵素連結免疫斑點法檢測顯示,免疫治療組的小鼠能有效引起周邊血液中抗原特異性CD8+ T細胞免疫反應。但以上的抗腫瘤特異性免疫反應在三組免疫治療組中均沒有明顯的差別(P >0.05)。總體來說,這三種異種異體重組多抗原結合位之免疫治療劑是安全的,並且具有同時引此B16-F10黑色素瘤小鼠身上的抗原特異性之細胞及體液免疫反應的潛在效力。 英文摘要 Differentiation antigens that can induce cytotoxic T cell, antibody, and antitumor response were found in melanoma cells. Three DNA plasmids expressing different multiepitopes were constructed; the ME plasmid expressing multiepitopes of gp100 (280V), gp100 (210M) and tyrosinase (370D), the MA plasmid expressing multiepitopes (ME) and full-length of Melan-A (MA), the MAGE plasmid expressing ME, MA and 2 copies of MAGE proteins. The aim of this study was to compare these three DNA immunotherapeutics in the B16-F10 murine melanoma model. Initially, protein expression of these three plasmids were confirmed by Western blot assay and fluorescent microscopy. The tumor-free interval, the tumor growths, and the metastasis rate were significantly different between the treatment groups (ME, MA, and MAGE groups) and the control groups (PEGFP and PBS groups). In addition, antigen-specific antibody responses were analyzed by Western blot assay in 16 out of 18, 12 out of 12, and 6 out of 6 immunized mice for ME, MA, and MAGE protein, respectively. The results of ELISPOT demonstrated that the number of antigen-specific CD8+ T cell in the treatment group were significant increases compare to control groups but there were no significant differences among the treatment groups. In conclusion, these three recombinant immunotherapeutics are safe, and able to stimulate tumor-specific effective immune responses.
URI: http://hdl.handle.net/11455/94758
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