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将骨髓细胞预先用抗人全部T细胞的单克隆抗体P81(MeAb P81)处理,再将这种处理过的骨髓细胞悬液通过兔抗小鼠IgG—Sepharose 6MB亲和层析柱。由于兔抗小鼠IgG抗体与MeAb P81包被细胞的特异性结合,将P81包被细胞滞留在柱内,使骨髓细胞中免疫活性细胞被清除。本文比较了三种不同温度对亲和层析柱分离效率的影响;观察了上柱的骨髓细胞数;同时摸索了P81的用量。清除率用间接免疫荧光法检测。实验表明,用亲和层析柱去除骨髓中成熟T细胞的最佳条件为:10~7个骨髓细胞,经20ml 1:500倍稀释的MeAb P81孵育,在4℃条件下,可使骨髓中免疫活性细胞的清除率达82±3.2％。用CFU—e测定表明细胞通过柱体后对骨髓粒系母细胞并无损伤。此法操作简便,省时,不需要昂贵的设备,重复性好,柱体亦可反复使用,可能有临床应用的潜力。
Bone marrow cells were pre-treated with monoclonal anti-human T cell monoclonal antibody P81 (MeAb P81), and the treated myeloid cell suspension was passed through a rabbit anti-mouse IgG-Sepharose 6 MB affinity column. Due to the specific binding of the rabbit anti-mouse IgG antibody to the MeAb P81-coated cells, the P81-coated cells were retained in the column and the immunocompetent cells in myeloid cells were cleared. In this paper, the effect of three different temperatures on the separation efficiency of affinity chromatography was compared. The number of myeloid cells on the column was observed. The amount of P81 was also explored. Clearance rate was detected by indirect immunofluorescence. Experiments show that the affinity chromatography column to remove bone marrow mature T cells in the best conditions: 10 ~ 7 bone marrow cells, 20ml 1: 500 dilution of MeAb P81 incubated at 4 ℃ conditions, can make the bone marrow Immunocompetent cells clearance rate of 82 ± 3.2%. The results of CFU-e assay showed that there was no damage to myeloid granulosa cells after the cells passed through the column. This method is simple, time-saving, does not require expensive equipment, good reproducibility, the cylinder can be reused, may have the potential for clinical application.