摘要
建立一种特异快速地诱导扩增人外周血γδT细胞的方法。用 5～ 10ml外周血通过CD3细胞的富集、γδ单抗的诱导以及肿瘤细胞刺激的培养体系进行γδT细胞的体外扩增。结果显示 ,γδT细胞能在短时间内快速大量扩增到 10 9～ 10 10 的数量级 ;诱导 4周的CD8、γδ阳性的细胞百分率分别为 42 6 9%和 5 8 6 7% ,明显高于诱导前的 2 3 97%和 6 6 2 % ;γδT细胞对NK敏感的K5 6 2细胞和NK抵抗的Daudi细胞均有较高的杀伤活性 ;γδT细胞对经抗HSP单抗封闭后的Daudi细胞的杀伤率明显下降。该培养体系是一种用血量少、特异、经济、快速的人外周血γδT细胞体外扩增方法。
Although γδT cells make up no more than 10% of the peripheral blood mononuclear(PBM)cells,they appear to play an important role in host defense against tumor growth.In order to evaluate their functional activity against tumor and clinical adoptive immunotherapy,large number of γδT cells are required.In this paper,we describe a simple and rapid method for the isolation and expansion of γδT cells from the peripheral blood.The method for purification and expansion from only 5～10ml of peripheral blood using solid phase anti CD3 antibody,anti γδ antibody and irradiated Daudi Burkitts lymphoma cells. Experimental results revealed: First,10 9～10 10 γδT cells could be gathered using this approach in about two weeks.Second,after 4 weeks,the percentages of CD8 and γδT cells in the expanded cells reached 42 69% and 58 67%,compared with 23 97% and 6 62% before expanding respectively. Third,after 4 weeks,purified γδT cells were shown significant cytotoxic activity against K562 and Daudi cells.Fourth,the cytotoxic activity against Daudi cells of γδT cells could be blocked by anti HSPantibody.This method is simple,special and rapid one for expanding and purification of γδT cells form 5～10ml of peripheral blood in vitro.
出处
《上海免疫学杂志》
CSCD
北大核心
2000年第1期47-49,共3页
Shanghai Journal of Immunology
