臺灣博碩士論文加值系統

雖然不斷有新的藥劑應用於治療氣喘疾病，在過去二十年間氣喘疾病的發生率仍一直在增加當中。現在的藥物雖能提供疾病症狀的部分舒解，但目前仍企需開發更具專一性的藥劑來遏止過敏性氣喘的發炎反應產生。因此，本研究宗旨即研發IL-12對家塵過敏原Der p 1所引發氣喘發炎動物模式免疫調節機制的探討。首先，利用小鼠建立起家塵過敏原Der p 1誘發的氣管發炎的動物模式，以提供進一步研究IL-12 在生物體的反應。在初步的研究中發現：當用過敏原Der p 1致敏老鼠，初期時即以IL-12注射老鼠時，不僅可促進老鼠體內Th1細胞的免疫反應，亦可抑制由呼吸道給予過敏原致敏所造成的呼吸道發炎現象。更進一步的，我們探討當經IL-12治療的老鼠是否仍可承受多次的過敏原致敏而不引發氣喘症狀的產生；以及測試那些經過敏原致敏導致Th2細胞反應而病發的老鼠，是否仍能因後期接受IL-12注射後而對於病情有改善的現象。由研究資料顯示：IL-12仍可用於治療已病發的老鼠，使呼吸道發炎症狀減輕。然而，另一結果顯示：在老鼠致敏初期注射IL-12並不能永久有效預防過敏性氣喘的發生。此外，為能更進一步的將IL-12應用在基因治療法上，我們利用了基因工程的方法，除了將組成IL-12蛋白質的p35單元體基因及p40單元體基因分別鑲嵌入同一真核載體外，亦另外將設計好的P35和p40組合成的融合基因一起鑲嵌入真核表現載體中。這兩個構築好的真核表現載體分別以pIL-12及pscIL-12代號稱呼之。測試這些質體在生物體內及體外的活性，發現pscIL-12質體不但確實能表現出具有生物活性功能的IL-12融合蛋白質，而且不必擔心有p40單元體的干擾。因此，將來利用pscIL-12質體來進行對過敏疾病的治療可能不失為一更適合的治療方式。

The incidence of asthma has increased substantially in the last two decades, despite increased variety of therapeutic agents. Existing drugs offer partial relief of symptoms in such disease, but there is a great need to develop more specific agents to suppress the inflammatory response in allergic asthma. In this study, we develop a novel therapeutic approach for Der p 1-induced allergic asthma with IL-12 treatment. First, an animal model of airway inflammation induced by house dust mite Der p 1 allergen was established. Then, the in vivo effects of IL-12 on the murine model of asthma were examined. The data demonstrated that IL-12 treatment during antigen sensitization not only promoted Th1 immune responses but also inhibited airway inflammation after a single inhaled antigen challenge. Moreover, the studies were extended to investigate the effects of IL-12 administration to maintain immune responses after multiple allergen challenge and to redirect immune responses after Th2-dominated conditions. The data demonstrated that IL-12 was useful as a therapeutic agent to abolish eosinophil recruitment, despite Th2-associated responses were existed. However, IL-12 could not prevent allergic asthma to maintain a stable and dominated Th1 responses for a long-term time. Furthermore, to apply IL-12 gene in gene therapy, plasmid expressing vectors, expressing coordinately p35 and p40 subunits or a single-chain IL-12 protein, was constructed and designated as pIL-12 or pscIL-12, respectively. The bioactivity of recombinant IL-12 was analyzed both in vitro and in vivo studies. These data revealed that pscIL-12 could encode functional single-chain IL-12 fusion protein and the free p40 subunits which can behave as IL-12 antagonist were negligible. Thus, such pscIL-12 may provide a more appropriate tool in future application of gene therapy for allergic diseases.

封面
中文摘要
英文摘要
誌謝
目錄
縮寫
Chapter I
1-1
1-2
1-3
1-4
1-5
1-6
1-7
Chapter 2 Animal model of airway inflammation induced by house dust mite Der p I allergen
2-2
2-3
2-4
2-5
Chapter 3 Administration of IL-12 prevents mite Der p I allergen-lgE antibody production and airway eosinophils infiltration in animal model of airway inflammation
3-1
3-2
3-3
3-4
3-5
Chapter 4 Administration of interleukin-12 exerts a therapeutic instead of a long-term preventive effect on mite Der p I allergen induced animal model of airway inflammation
4.1
4-2
4-3
4-4
4-5
Chapter 5 Construction of vectors expressing bioactive heterodimeric and single-chain murinem interleukin-12 for gene therapy
5-1
5-2
5-3
5-4
5-5
Chapter 6 Discussion and future work
6-1
6-2
6-3
6-4
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