Tsoi M S, Aprile J, Dobbs S, Goehle S, Storb R
J Immunol Methods. 1982 Sep 30;53(3):293-305. doi: 10.1016/0022-1759(82)90176-4.
A cell separation method using immunoglobulin (Ig)-coated plates, originally devised for murine spleen cells, was modified and adapted for enrichment (and depletion) of cellular subpopulations from human peripheral blood. For the direct separation of B and T cells, F(ab')2 fragments of anti-human Ig were used to coat the plates. For indirect separation, the cells were first incubated with monoclonal antibodies to cell surface antigens and then separated in plates coated with anti-mouse Ig. Plates were first coated with poly-L-lysine to facilitate the adherence of anti-Ig antibodies, and finally with bovine serum albumin to mask free poly-L-lysine. Cells which did not react with the anti-Ig antibodies or which were nonadherent to the plate were pipetted off; cells which reacted with the anti-Ig antibodies or which were adherent were eluted after incubation with excess serum. T, non-T, T4+, T4-, T8+, and T8- lymphocytes were separated with high viability, purity, and yield. The method was used to study suppressor activity of a patient who was treated by bone marrow transplantation for myelofibrosis. Strong suppressor activity was associated with unfractionated peripheral blood mononuclear leukocytes, monocytes, T, T8+, and T4- cells but not with B, T8-, and T4+ cells.
一种最初为小鼠脾细胞设计的使用免疫球蛋白（Ig）包被板的细胞分离方法经过改进，适用于从人外周血中富集（和去除）细胞亚群。为了直接分离B细胞和T细胞，使用抗人Ig的F(ab')2片段包被板。为了间接分离，细胞首先与细胞表面抗原的单克隆抗体孵育，然后在抗小鼠Ig包被的板中分离。板首先用聚-L-赖氨酸包被以促进抗Ig抗体的粘附，最后用牛血清白蛋白覆盖游离的聚-L-赖氨酸。不与抗Ig抗体反应或不粘附于板的细胞用移液管吸出；与抗Ig抗体反应或粘附的细胞在与过量血清孵育后洗脱。T细胞、非T细胞、T4+细胞、T4-细胞、T8+细胞和T8-细胞以高活力、纯度和产量被分离。该方法用于研究一名接受骨髓移植治疗骨髓纤维化患者的抑制活性。强抑制活性与未分级的外周血单个核白细胞、单核细胞、T细胞、T8+细胞和T4-细胞有关，而与B细胞、T8-细胞和T4+细胞无关。
