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National Chung Hsing University Institutional Repository - NCHUIR > 獸醫學院 > 獸醫學系所 > 依資料類型分類 > 碩博士論文 >  表現異種異體黑色素分化抗原之DNA對於黑色素瘤小鼠的抗腫瘤免疫反應

Please use this identifier to cite or link to this item: http://nchuir.lib.nchu.edu.tw/handle/309270000/155282

標題: 表現異種異體黑色素分化抗原之DNA對於黑色素瘤小鼠的抗腫瘤免疫反應
Antitumor immune responses in melanoma-bearing mice received DNA expressing xenogeneic melanosomal differentiation antigens
作者: 徐子涵
Hsu, Tz-Han
Contributors: 張仕杰
Shih-Chieh Chang
獸醫學系暨研究所
關鍵字: 黑色素瘤小鼠;異種異體;DNA疫苗;抗原特異性免疫反應;抗腫瘤效力
Melanoma-bearing mice;Xenogeneic;DNA vaccine;Antigen-specific immune response;Antitumor efficacy
日期: 2012
Issue Date: 2013-11-21 11:49:19 (UTC+8)
Publisher: 獸醫學系暨研究所
摘要: 黑色素瘤細胞所表現的分化抗原(例如Melan-A、tyrosinase及gp100)已證實能引起毒殺性T細胞及抗體反應。本實驗室先前架構出一種同時帶有gp100 (280V)、gp100 (210M)及tyrosinase (370D)之異種異體多抗原結合位的DNA免疫治療劑。本實驗的研究目的為評估此多抗原結合位免疫治療劑,對於B16-F10黑色素瘤小鼠的抗原特異性免疫反應及抗腫瘤效力。首先,以肌肉注射方式給予黑色素瘤小鼠10 (ME 10組)、50 (ME 50組)及250 μg (ME 250組)三種不同劑量之多抗原結合位質體DNA,一週一次,共四次。結果發現ME 50及250組有顯著的抑制腫瘤生長效果(P < 0.05)。而酵素連結免疫斑點法檢測顯示,ME 250組能有效引起周邊血液中抗原特異性CD8+ T細胞免疫反應。此外酵素連結免疫吸附法及西方墨點法分析顯示,在17隻給予多抗原結合位質體DNA中的7隻小鼠發現有抗原特異性抗體反應。而存活分析試驗中發現,ME 250組的存活時間比起對照組顯著延長(P < 0.05)。總結,本研究發現異種異體重組多抗原結合位之免疫治療劑是安全的,並具有同時引起抗原特異性之細胞及體液免疫反應的潛在效力,而此抗腫瘤免疫效力能抑制B16-F10黑色素瘤生長且延長小鼠存活率及存活時間。
Melanoma cells have been found to express differentiation antigens (e.g., Melan-A, tyrosinase and gp100) that can induce cytotoxic T-cell and antibody responses. Previously, the DNA-based immunotherapeutics with genes encoding xenogeneic multiepitope (ME) comprising of gp100 (280V), gp100 (210M) and tyrosinase (370D) was constructed in our laboratory. The goal of this study was to evaluate the antigen-specific immune responses and antitumor efficacy of the ME immunotherapeutics in the B16-F10 murine melanoma model. Melanoma-bearing mice were intramuscularly injected with ME plasmid DNA in various doses including 10 (ME 10 group), 50 (ME 50 group) and 250 μg (ME 250 group) weekly for a total of four injections. Results demonstrated that significant inhibition of tumor growth was found in the ME 50 and 250 groups (P < 0.05). Antigen-specific CD8+ T-cells in peripheral blood were detected by enzyme-linked immunospot assay in all the mice of the ME 250 group. In addition, antigen-specific antibody responses were analyzed by both enzyme-linked immunosorbent assay and Western blot assay in 7 out of 17 immunized mice. The survival times of the ME 250 group were significantly longer than that of control groups (P < 0.05). In conclusion, the findings of this study demonstrated that xenogeneic recombinant ME immunotherapeutics was safe and was potential to elicit both antigen-specific cellular and antibody immune responses. This antitumor immunity had an inhibitory ability against the B16-F10 melanoma growth to improve the survival rate and survival time of mice.
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