Development and Application of Monoclonal Antibodies to Peripheral Blood Monocyte Derived Macrophages of Hybrid Tilapia (Oreochromis niloticus × Oreochromis Mossambicus)
| Autor: | Yu-Jung Hsieh, 謝育錚 |
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| Rok vydání: | 2008 |
| Druh dokumentu: | 學位論文 ; thesis |
| Popis: | 96 The aim of this study is to develop and apply the monoclonal antibodies (MAbs) to the macrophages from hybrid tilapia (Oreochromis niloticus × Oreochromis Mossambicus) because there are a limited number of MAbs to fish leukocytes have been described until today. Our study results show that we are able to get eight MAbs, mAbM1~mAbM8, produced from Balb/c mice immunized with peripheral blood monocyte derived macrophages (PBM) of hybrid tilapia. Those mAbs have been screened from a lot of hybridoma clones by indirect ELISA and analyzed with macrophages from peripheral blood leucocytes (PBL). The result reveals that the antibody titers of mAbM1, mAbM2, mAbM6 and mAbM7 are higher than the others. Those four MAbs reacted with PBM (peripheral blood macrophages), HKM (head kidney macrophages) and SM (spleen mcrophages) are used to be analyzed their characterizations by the indirect immunofluorescence assay test (IIFAT) and flow cytometry (FCM). The results suggest these four MAbs have very high activities with PBM and HKM, especially mAbM6 and mAbM7. Furthermore, mAbM6 and mAbM7 are chosen for testing the cross-reactivity with peripheral blood monocyte derived macrophages from different species including eel, carp, mice, rabbit, chicken, sheep, dog, and human. The results indicate that both mAbM6 and mAbM7 have significantly higher reactivity with PBM from eel, mice, sheep and chicken. Interestingly, the capping or ring forms of the mAbM6＋ and mAbM7＋ cells are recognized and found by the confocal microscope. Therefore, we presume that both MAbs binding epitopes are on the cell surface of the macrophages. Finally, we find that our developed mAbM6 and mAbM7 MAbs could be sorted the macrophages from peripheral blood leukocytes successfully. Indeed, our MAbs could serve as a useful tool to identify fish macrophages for further studies. |
| Databáze: | Networked Digital Library of Theses & Dissertations |
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