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題名 | 以B型肝炎表面抗原離體免疫人類B淋巴球 |
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作 者 | 黃俊奇; 周千慧; 曾哲明; | 書刊名 | 師大生物學報 |
卷期 | 26 1991.12[民80.12] |
頁次 | 頁11-19 |
分類號 | 414.82 |
關鍵詞 | B型肝炎表面抗原; B淋巴球; 人類; 離體免疫; HBsAg; In vitro immunization; |
語文 | 中文(Chinese) |
中文摘要 | B型肝炎的感染與帶原者的比率,在台灣地區仍是非常高,而目前預防與治療方面的研究方向之一,是製造出人類抗B型肝炎表面抗原(HBsAg)的單株抗體。製造單株抗體首要步驟是以離體免疫法增加抗HBsAg抗體製造細胞。報告指出在離體免疫以前,以Leu-OMe去除週進血液單核細胞(PBMN)中之富含溶小體細胞(lysosome-rich cells)或以OKT-8加補體去除CD8+T細胞,可增加人類淋巴球對多種抗原離體免在之效率;本實驗在探討同樣的處理,是否可促進HBsAg離體免疫人類B淋巴球的效率,而增加抗HBsAg抗體製造細胞的數目,再以Filter Immunoplaque assay (FIPA)測定抗體製造細胞的數目。實驗結果顯示,處理過Leu-OMe或OKT-8加補體,6天後,抗HBsAg抗體製造細胞的數目,顯著增加,而Leu-OMe或OKT-8加補體並不影響離體免在過程中,其他免疫細胞之存活率。經Leu-OMe處理後之淋巴球對淋巴球活化劑(PWM: Pokeweed mitogen)之反應,並不隨PWM濃度增加而增加抗HBsAg抗體製造細胞數目,但是其最佳濃度為10ug/ml。總之,經Leu-OMe或OKT-8加補體處理PBMN後,應可對隨後之製造人類抗HBsAg單株抗體有所助益。 |
英文摘要 | The researches on prophylaxis and diagnostics of Hepatitis B (HB) are still in the first priority due to the high percentage of chronic HB virus carrier in Taiwan. The current trend in HB research is to generate the human monoclonal antibodies against HB surface antigen (HBsAg), which is generally initiated by an in vitro immunization process for increasing the number of antigan-specific antibody forming cells. Several reports demonstrated that depletion of lysosome-rish cells by Leu-OMe or elimina-tion of CD8 + T lymphocytes by OKT8-mediated complement reaction resulted in an increase in the efficiency of in vitro immunization. In this report, we examine the ef-fect of the same treatments on the efficiency of HBsAg-mediated in vitro immuniza-tion by measuring the number of spot-forming cells (SFC) on the filter of a Filter Immunoplaque Assay (FIPA). The data indicated that depletion of Leu-OMe sensi-tive cells or OKT8+ cells from peripheral blood mononuclear cells (PBMN) signifi-cantly increase the number of SFC after six days of in vitro immunization. The via-bility of the rest of immune cells were not affected by the treatments during the sub-sequent immunization period. After Leu-OMe treatment, the cells responded nor-mally to pokeweed mitogen (PWM), a nonspecific B cell activator with an optimal dose at 10ug PWM per ml. therefor, our studies suggested that depletion of lysosome-rich or CD8+ cell population did enhance the number of HBsAg-specific antibody forming cells, thus would increase the probability of cloning out the anti-HBsAg anti-body producer. |
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