Hoshino T, Hara A, Inoue M, Honda J, Imai Y, Oizumi K, Yokoyama M M
Department of Immunology, Kurume University School of Medicine, Fukuoka, Japan.
J Clin Lab Immunol. 1991 Sep;36(1):39-43.
A simple and fast method to measure the natural killer (NK) cell cytotoxicity has been established. In our novel assay, following the incubation of K562 target and effector cells, the cells were stained with FITC-conjugated anti-transferrin receptor (CD71) mAb. The K562 targets alone were labeled with anti-CD71 mAb, since CD71 is highly expressed on K562 cells but not on mononuclear cells. The fluorescence intensity of CD71 on K562 cells were analyzed using flow cytometry, and the cytotoxicity was evaluated. In our assay, designated as a "CD71 assay", no spontaneous release of anti-CD71 mAb was observed from labeled K562 cells and living K562 cells were able to be divided from dead K562 cells. The data obtained by this assay was well correlated with that by the standard 51Cr-release assay as well as the C-FDA assay. In addition, the CD71 assay is a safe and simple analytic procedure since no isotope is required.
已建立一种简单快速的方法来测量自然杀伤（NK）细胞的细胞毒性。在我们的新检测方法中，将K562靶细胞和效应细胞孵育后，用异硫氰酸荧光素（FITC）偶联的抗转铁蛋白受体（CD71）单克隆抗体对细胞进行染色。单独的K562靶细胞用抗CD71单克隆抗体标记，因为CD71在K562细胞上高表达，而在单核细胞上不表达。使用流式细胞术分析K562细胞上CD71的荧光强度，并评估细胞毒性。在我们称为“CD71检测”的方法中，未观察到标记的K562细胞有抗CD71单克隆抗体的自发释放，并且活的K562细胞能够与死的K562细胞区分开来。通过该检测获得的数据与标准的51Cr释放检测以及C-FDA检测的数据高度相关。此外，CD71检测是一种安全简单的分析方法，因为不需要同位素。
