磁柱分选技术在CMV特异性CD8^+ T细胞纯化中的应用研究  

Study on the purification of CMV-specific CD8^+ T cells by using the technology of MS column

作　　者：王新超[1,2] 张伟[2] 邸旭[2] 姜智[3] 陈宝安[1] 

机构地区：[1]东南大学医学院,江苏南京210009 [2]天津医科大学四中心临床学院肿瘤外科,天津300140 [3]江苏省肿瘤医院妇瘤科,江苏南京210009

出　　处：《东南大学学报（医学版）》2010年第4期413-416,共4页Journal of Southeast University(Medical Science Edition)

基　　金：国家高技术研究发展计划(863计划)项目(207AA022007)

摘　　要：目的:探索联合磁选技术和巨细胞病毒(CMV)连锁状多聚体磁珠纯化CMV特异性CD8+T细胞,从而用于CMV疾病的过继性免疫治疗。方法:运用CMV连锁状多聚体MHC-磁珠标记CMV特异性CD8+T细胞,然后用MS磁柱分选,从HLA-A2/CMV血清阳性健康志愿者的外周血标本中纯化CMV特异性CD8+T细胞。为了优化分选效果,分别将标记后的标本进行1次和两次MS磁柱分选,比较纯化后的CMV特异性CD8+T细胞的纯度。结果:通过MS磁柱分选,CMV特异性CD8+T细胞的最高纯度达到90.45%。经过1次和两次MS磁柱分选后,CMV特异性CD8+T细胞的纯度有显著差异,但经过两次磁柱分选较1次分选的细胞丢失严重。结论:联合磁柱分选技术和CMV连锁状多聚体磁珠纯化CMV特异性CD8+T细胞效果良好,有利于对CMV疾病的过继性免疫治疗。Objective：To investigate whether the technology of MS column and the complex of cytomegalovirus（CMV） streptamer MHC-magnetic beads could offer the advantage of the purification of CMV-specific CD8 ＋ T cells,therefore benefit for the adoptive immunotherapy of CMV disease.Methods：CMV-specific CD8 ＋ T cells were selected by magnetic column from the peripheral blood of HLA-A2 /CMV sero-positive healthy donors by using the complex of CMV streptamer MHC-magnetic beads labeling.For best results,we compared the advantage of purification by using MS column between two columns selection and one column selection.Results：After positive selection by MS column,a purity with up to 90.45% was achieved for CMV-specific CD8 ＋ T cells.The frequency of CMV-specific CD8 ＋ T cells had a significant difference on the purity of selected T cells when purified by different times of MS column selection,but the loss of CMV-specific CD8 ＋ T cells was severe when purified by two columns selection.Conclusion：It is concluded that the technology of MS column and the complex of CMV streptamer MHC-magnetic beads could be used to purify CMV-specific CD8 ＋ T cells for adoptive immunotherapy.

关 键 词：MS磁柱 连锁状多聚体 巨细胞病毒 获得性免疫治疗 T细胞 

分 类 号：R373.9[医药卫生—病原生物学] R457.2[医药卫生—基础医学]