共表达膜型IL-15和RAE的细胞克隆的筛选和鉴定  

Preparation and identification of a monoclonal cell for co-expression of mouse membrane-bound form of IL-15 and RAE

作　　者：钱莉[1] 陆家辉[1] 潘兴元[1] 田芳[1] 龚卫娟[1] 季明春[1] 

机构地区：[1]扬州大学医学院病原生物学与免疫学教研室,江苏225001

出　　处：《齐齐哈尔医学院学报》2011年第9期1367-1368,共2页Journal of Qiqihar Medical University

基　　金：国家自然科学基金(81001308);江苏省自然科学基金(BK2010315)

摘　　要：目的筛选能够共表达膜型IL-15和RAE的细胞株,为扩增和活化特定免疫细胞提供有效的实验工具。方法前期工作中已通过潮霉素筛选获得共表达膜型IL-15和RAE的BaF3细胞,但阳性细胞所占的比例较低。本实验通过有限稀释法先筛选高表达RAE的单克隆细胞株,再筛选高表达膜型IL-15的单克隆细胞株,最终获得了1株共表达膜型IL-15和RAE的单细胞克隆。将该细胞株与NK细胞共培养后,通过IFN-γ的分泌验证其生物学活性。结果流式细胞仪证实该细胞株能够高水平共表达膜型IL-15和RAE。并具有很强的活化NK细胞的效应。结论成功地制备了共表达膜型IL-15和RAE的单克隆细胞株,为活化特定免疫细胞,如IKDC和NK细胞提供实验工具。Objective To prepare and identify a monoclonal cell that co-expressed of mouse membrane-bound form of IL-15（mb15） and RAE. Methods BaF3 cell that co-expressed of mouse mbIL-15 and RAE was cloned by limiting dilution.Flow cytometry（FCM） analysis was then used to identify the clone.Moreover,we analyzed the IFN-γ secretion by NK cells after their interaction with BaF3/mb15/RAE cells for 24 h.Results The cell clone with the highest expression level of RAE and mb15 was selected and it could promote the biological function of NK cells manifested as induced IFN-γ production.Conclusions Monoclonal cell that co-expressed of mb15 and RAE has been prepared successfully and can be served as a useful tool for the proliferation and activation of certain subsets of lymphocytes,such as NK cells and IKDCs in vitro.

关 键 词：IL-15 RAE 克隆细胞 

分 类 号：R392.11[医药卫生—免疫学]