石斑魚為本地重要經濟魚種，為了進一步應用免疫反應控制疾病，相關檢測工具也需要進一步的研發。本論文旨在說明如何在新物種，未知免疫基因的狀況下，篩選出合適免疫細胞標示蛋白應用於免疫研究。論文中的標記蛋白主要透過受到LPS免疫刺激以及沒有受到刺激的白血球所萃取合成的cDNA透過扣減雜合反應後所篩選出來與免疫相關的候選蛋白。第一個細胞標的為 CD53，CD53屬於tetraspanin的一員，發現 CD53 專一性表現在類白血球細胞上，因此可以做為石斑魚血球的標記。同時我們也發現石斑魚受到免疫刺激時，CD53 基因表現量有下降的趨勢，顯示與石斑魚的免疫反應可能有相關。另一個細胞標示基因為 CCL4，CCL4 為趨化素的一員，受到免疫刺激時會有非常高量的表現，因此分析血液中 CCL4 的表現量，可以監控養殖場內的魚隻是否有因為病原入侵而誘發非特異性免疫反應。在本論文中除了這兩個分子的應用分析外，此二分子在免疫上可能扮演的角色亦被進一步的探討，將發現的結果運用在增加石斑魚的免疫力及降低發炎所產生的傷害，做為提升石斑魚整體的產量方法。

Identifying immune cell markers and developing appropriate tools (such as antiserums) to detect those markers can greatly benefit the study of a species’ immune response. Grouper (Epinephelus spp.) is an important economic fish species in Taiwan. In order to control disease outbreak in grouper aquaculture, better understanding of the grouper immune system is needed. However, a good marker of immune system is important. In this thesis, we describe how to screen for and select appropriate markers for a novel species. We also prepare novel immune tools to benefit immunology research. Candidate protein markers were selected. The first marker we identified is CD53, a tetraspanin. Findings from this research revealed thatCD53 is highly expressed in the leukocyte-like cells of immune organs, which suggests that CD53 can be used as a marker to identify leukocytes. Moreover, CD53 expression decreased in LPS stimulated groupers, which further confirms that CD53 is related to grouper immune response. In contrast to CD53, the grouper CCL4 gene was highly expressed following LPS treatment. Specifically, CCL4 is a chemokine which is highly expressed in groupers after treatment and can potentially be used to help monitor fish health. The biofunctions of CD53 and CCL4 markers were also evaluated, and their functional roles were found to differ in grouper fish. The results can be found in the application of the increase in the grouper immune system and reduce the harm caused by inflammation, then as a policy to increase the production of groupers.

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