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頁籤選單縮合
題名 | Separation of Penaeus Monodon Hemocytes by Negative Selection=利用負篩選分離草蝦的血球細胞 |
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作者 | 宋宏紅; 孫咸義; Sung, Hung-hung; Sun, Ryan; |
期刊 | 臺灣水產學會刊 |
出版日期 | 20001200 |
卷期 | 27:4 2000.12[民89.12] |
頁次 | 頁295-301 |
分類號 | 381.8 |
語文 | eng |
關鍵詞 | 血球的分離; 草蝦; 單株抗體; 負篩選; Hemocyte separation; Penaeus monodon; Monoclonal antibody; Negative selection; |
中文摘要 | 不族群血球細胞的分離,對草蝦晚疫性細胞之影響因子的研究是必要的。本研究利 用四株具有辨識草蝦血球細胞的專一性單株抗體,將其分別連結到磁後,當做工具以負篩選 方法進行不同次族群血球細胞的分離。晚疫金染色與免疫螢光染色的結果顯示,每一種次族 群中(包括透明球、半顆粒球及顆粒球)都有一些血球細胞能分別被這四種抗體標幟,且所有 被辨識的抗原都同時出現在細胞表面及細胞質內的顆粒與細胞核中。利用連接單株抗體的磁 以單一負篩選(single selection)後,血球樣本的透明球密度會增加至90%;利用連接Z6A5或 Z6H8的磁珠處理,透明球甚可高達95%。然而,利用重負篩選(double selection)增加的透明 球密度程度與單一負篩選無差異。 |
英文摘要 | In order to study immune cellular effectors of tiger shrimp (Penaeus monodon), it is necessary to separate hemocytes into different subpopulations. In this study, four monoclonal antibodies (mAbs) against tiger shrimp hemocytes were conjugated individually to magnetic beads as tools to separate hemocytes via negative selection. Results from the immunofluorescent and immunogole stains showed that each subpopulation of hemocytes including hyaline cells (HC), semigranular cells (SGC) and granular cells (GC)) could be separately labeled by the four mAbs as followed: 13% HC, 23% SGC and 6% GC labeled by Z5E10, 4% HC, 7% SGC and 1% GC labeled by Z6A5, 6%HC, 21% SGC and 3% GC labeled by Z6A6, and 5% HC, 3% SGC and 3% GC labeled by Z6H8. Percentages of total hemocytes recognized by Z5E10, Z6A5 and Z6H8 were 41%, 12%, 30% and 11%, respectively. The Z6A5- or Z6H8-negative hemocytes whowed even higher percentage, primarily HC (74% and 73%, respectively). However, mAb Z5E10 mainly reacted with granulocytes (including SGC and GC). The mAb-positive hemocytes observed by using a transmission electron microscope (TEM) via immunoglod staining showed that all of the antigens recognized by the four mAbs simultaneously present on the surface of cells and in cytoplasmic granules and cell nucleus. All of the antigens recognized by the four mAbs were indentically presented on the cell surface and in granules and nucleus of hemocytes. Folloing single-negative selection, the HC density in hemocyte samples treated with each mAb was increased higher than 90%, or even up to95% when samples treated by Z6A5- or Z6H8-neads. However, the incressing level of HC density showed on difference between samples treated by single negative or double negative selection. |
本系統之摘要資訊系依該期刊論文摘要之資訊為主。