用抗人T淋巴细胞单克隆抗体去除骨髓中成熟T细胞的影响因素

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。

FACTORS IN DEPLETING MATURE T LYMPHOCYTES FROM BONE MARROW CELLS BY McAb

A monoelonal antibody against T lymphocyles, designated as McAb P81, has been used to separate T lymphocytes from bone marrow cells (BMC). After incubation with McAb P81, the cells were added to a Rabbit antimouse (RAM)-Sepharose 6MB affinity chromatoraphy column, which was made by coupling lg of CNBr-Sepharose 6MB with 20mg of RAM IgG. T cells were specifieally bound to the column and non-bound cells Were eluted. The efficiency of depleting of T cells from BMC was compared under different conditions as temperature ranges, numbers of BMC added to the Column and different McAb P81 concentrations used. It was observed that all the three factors mentioned above were the major effectors on the results of T-cell depleting. Under the optimal condition which we found in incubating BM cells with 20 μl McAb P81(1:500) at. 4℃ the elimination rate of immunocompetent cells from BMC was 82.0％ (S.D.±3.2％), the viability of which was over 95％. Using CFU-cin detection revaled no injury to the eluted cells.