T淋巴细胞清除促进脐血造血细胞的体外扩增

Elimination of T Lymphocytes Promotes the Expansion of Hematopoietic Cells in vitro

采用抗CD3或抗CD8单克隆抗体的补体细胞毒方法清除脐血单个核细胞 (MNC )中T淋巴细胞 ,CD34免疫亲和柱纯化MNC中CD34 +细胞 ,流式细胞技术 (FACS )分析细胞表面标志。将CD34 +细胞中加入含多种造血生长因子的培养基进行体外液态扩增 ,并观察粒 巨噬集落 (CFU GM )和多向祖细胞集落 (CFU GEMM )形成能力。结果CD3+或CD8+细胞清除组和MNC经CD34免疫亲和柱纯化后 ,CD34 +细胞分别为 5 9 5 2 %、 5 6 70 %和 5 0 72 % ,比未纯化组 (1 0 7% )纯度大幅度提高。抗CD3单抗清除 +CD34纯化组和单纯CD34纯化组经造血生长因子刺激培养第 14天 ,细胞总数分别扩增 110 40倍和 87 0 0倍。抗CD3单抗清除 +CD34纯化组、抗CD8单抗清除 +CD34纯化组和单纯CD34纯化组的CFU GM产率分别为 2 86 5 0± 12 0 2、2 88 5 0± 17 68和 2 19 5 0± 5 3 0 3,前者虽高于后者 ,但差异无显著性。CFU GEMM产率分别为 376 67± 43 2 4、 438 33± 36 73和 311 0 0± 40 11,抗CD3单抗清除 +CD34纯化组无显著差异 ,抗CD8单抗清除 +CD34纯化组显示出显著性作用 (P <0 0 5 )。

The T lymphocytes in mononuclear cells (MNC) of cord blood were eliminated by using anti CD3 or anti CD8 monoclonal antibodies with complement as well as by purification of CD34 + cells with immunoaffinity columns,and cell surface antigens were analyzed by means of FACS The hematopoietic cells were expanded in a liquid cultural system with hematopoitic growth factors added,and the numbers of CFU GM and CFU GEMM were determined later on The experimental results showed that,after treatment with anti CD3 or anti CD8 monoclonal antibodies followed by immunoaffinity columns purifications The purities of CD34 + cells were 59 52% and 56 70% respectively,while those of the controls (those treated with immunoaffinity columns alone) was 50 72%. The total number of cells expanded by 110 40 and 87 00 folds in the group treated with anti CD3 monoclonal antibody and the control respectively after cultivation of cells for 14 days In these three groups of experiments,the numbers of CFU GM and those of CFU GEMM were 286 50±12 02,288 50±17 68,219 50±53 03 as well as 376 67±43 24,438 33±36 73,311 00±40 11 respectively There was significantly difference in the numbers of CFU GEMM between the groups treated with anti CD8 monoclonal antibody and the controls