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標題: | 改造人類自然殺手細胞株使其對表現癌胚抗原之腫瘤細胞具特異之毒殺作用 Retargeting of human natural killer cell line cytolytic activity to CEA-expressing cancer cells results in specific tumor cell destruction |
作者: | Kuo-Wei Huang 黃國維 |
指導教授: | 季昭華 |
關鍵字: | 崁合受器,癌胚抗原,自然殺手細胞, chimeric receptor,carcinoembryonic antigen,NK cell, |
出版年 : | 2006 |
學位: | 碩士 |
摘要: | NK92MI為一種可以快速生長,且可以自行分泌IL-2而不需額外補充的人類自然殺手細胞株,其擁有對多種腫瘤細胞具有毒殺的能力,但卻不會影響到正常的人類細胞,而也由於這些特性,使得此細胞被認為是發展免疫治療絕佳的工具之ㄧ。為了加強NK92MI對抗腫瘤的能力以及拓展其毒殺能力的範圍,在本實驗裡我們將一段崁合受器的基因序列,利用反轉錄病毒系統,將其送入NK92MI內表現。此崁合受器由對癌胚抗原具特異性的單鏈抗體、人類CD8 α hinge region及人類CD3ζ chain transmembrane和intracellular domain所構成。結果在體外細胞毒殺測試方面,可觀察到NK92MI與改造後的NK92MI皆無法對沒有表現癌胚抗原的腫瘤細胞進行毒殺作用;然而實驗發現,原本NK92MI無法毒殺的對象,若其有表現癌胚抗原,則即使在低的作用細胞與目標細胞比例下,改造後的NK92MI即可展現對此細胞高效率且特異的毒殺作用。因此在本實驗中,我們利用崁合受器在NK92MI上的表現,成功的改變了其原本的毒殺能力,而使得其對於表現癌胚抗原的細胞可以展現強大且特異的毒殺作用。若此研究可以持續發展推動,未來有極大的潛力與機會可以用於治療有大量癌胚抗原表現的腫瘤上。 The continuously growing and IL-2-independent natural killer (NK) cell line NK92MI is highly cytotoxic against malignant cells of various origins without affecting normal human cells. These properties suggest that NK92MI may be a good candidate as an immunotherapeutic agent. To further enhance the antitumoral activity of NK92MI cells and expand the range of tumor entities suitable for NK92MI–based therapies, here by transduction with a retroviral vector we have generated genetically modified NK92MI cells expressing a chimeric antigen receptor (CAR) specific for the tumor-associated carcinoembryonic antigen (CEA), which is overexpressed in many tumors of epithelial origin. The CAR consists of the CEA-specific single chain antibody (scFv), a flexible hinge region derived from CD8, and transmembrane and intracellular regions of the CD3ζ chain. Transduced NK92MI-scFv (CEA) -ζ cells express significant levels of the fusion protein on the cell surface as determined by fluorescence-activated cell-scanning (FACS) analysis. In cytotoxicity assays, there was no difference in cytotoxic activity of NK92MI and NK92MI-scFv (CEA) -ζ cells toward CEA-negative targets. However, even at low effector-to-target ratios, NK92MI-scFv (CEA) -ζ cells specifically and efficiently lysed CEA-expressing tumor cell lines that were resistant to cytolytic activity of parental NK92MI cells. These results demonstrate that efficient retargeting of NK92MI cytotoxicity can be achieved and might allow the generation of potent cell-based therapeutics for the treatment of CEA-expressing malignancies. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/32335 |
全文授權: | 有償授權 |
顯示於系所單位: | 獸醫學系 |
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