魚類後天性免疫系統如同哺乳類可提供長期、高效且具記憶性之保護力，
亦可分為體液型及細胞媒介型免疫反應且由 B cell 及 T cell 合作調節。
T cell 根據表面的 CD (Cluster of differentiation) 分子可分為 Helper T
cell (TH) (CD4+/CD8-) 及 Cytotoxic T cell (TC) (CD4-/CD8+)。TH 在抵抗
胞外菌入侵時傾向分化成 TH2 啟動 B cell 及抗體反應及受到胞內菌
感染時會分化成 TH1 啟動 TC cell 增生。因此疫苗若能調節 TH1 及
TH2 的分化將可增進疫苗效率。在實務上，疫苗設計與製造過程中經常
藉由添加佐劑用以增強或調節免疫反應，然而，需要一個方便有效的工
具來評估添加這些佐劑對於免疫系統的影響．目前魚用疫苗的體外效力
評估多以抗體力價作為特異性體液型免疫反應的指標，然而在細胞型免
疫反應則較為複雜且尚未有合適的評估方式。故本研究旨在建立以流式
細胞儀平台偵測點帶石斑魚免疫器官中表現特定 T cell 表面分子的細
胞率變化來分析免疫反應的走向。結果顯示，利用此方法可測得由疫苗
引起的 TH 及 TC 細胞分化反應，且證實在細胞激素的添加下可調節此
反應的走向。本研究的成果在未來可以提供一個技術平台，作為疫苗設
計、效力評估及推估適當的注射及追加時間點的研究工具。

Adaptive immunity in teleost provides a long term, highly effective and specific protection against future encounter with the same pathogens as its counterpart in mammals. It is divided into humoral and cell-mediated immunity and T cell and B cell collaborate with each other to regulate the entire adaptive immune system. T cell is divided into helper T cell (TH, CD4+/CD8-) and cytotoxic T cell (TC, CD4-/CD8+) by different CD markers expressed on cell surface. To combat extracellular pathogen, TH predominantly develop into TH2 cells to activate B cell response and antibody production while differentiate into TH1 cells to activate TC proliferation when infected with intracellular pathogens. Therefore, it is possible to improve vaccine efficacy via regulate TH1 or TH2 differentiation. Practically, the adjuvant is often used to enhance or modulate the immune response during the design and manufacture of the vaccine. However, a convenient and effective tool is needed to evaluate the effect of the addition of these adjuvants on the immune system. During the vaccine design, antibody titer is frequently used to represent humoral mediated immunity. However, methods to assess cell-mediated immunity are much more complex and still no appropriate tools can be applied. In this study, flow cytometry is an ideal method which can characterize multiple cell type by using cell markers, which could give a detail cell population profile in the immune organs, providing a better assessment of T cell response. Results showed that in Epinephelus coioides, increase of TH and TC population induced by vaccine were detected by flow cytometry and cytokines as vaccine additive were proved to enhance these responses. In the future perspective, this flow cytometry system can be further applied to vaccine design and optimize a vaccine schedule.

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