Masucci G, Lindemalm C, Frödin J E, Hagström B, Mellstedt H
Department of Oncology, Karolinska Hospital, S-104 Stockholm, Sweden.
Hybridoma. 1988 Oct;7(5):429-40. doi: 10.1089/hyb.1988.7.429.
Peripheral blood mononuclear cells (PBMC) from healthy individuals were studied for their lytic capability in ADCC using SW948 (a human colorectal carcinoma cell line) as target cells. Three monoclonal antibodies (MAbs) were used: two mouse MAbs (IgG2A) against the antigenic structures CO17-1A and BR55-2 respectively and one chimeric MAb 17-1A (IgG1) (mouse-human). Three kinds of effector cells were prepared. PBMC were purified on a Ficoll-Isopaque gradient (FIP cells) (a mixture of lymphocytes and monocytes). To obtain pure monocytes (greater than 90%), PBMC were centrifuged on a Nycodenz gradient (Nycodenz cells). Highly purified lymphocytes (greater than 98%) were obtained by treatment of FIP cells with iron powder and removal of phagocytic cells (PBL cells). Monocytes had the highest lytic capability. FIP cells were less effective than monocytes. PBL cells had the poorest killing activity. In reconstitution experiments addition of increasing amount of monocytes to PBL resulted in an augmented cytotoxicity. The numbers of Leu-M3+ cells, Leu-M5+ cells (monocytes) and CD16+ cells correlated positively to cytotoxicity. Higher concentration of MAb 17-1A was required to reach the same level of cytotoxicity using FIP cells as effector cells as compared to monocytes. MAb BR55-2 induced the same cytotoxic activity as MAb 17-1A. Combination of these two MAbs did not increase the lytic capability. Chimeric MAb 17-1A mediated ADCC in a dose-dependent fashion. The chimeric MAb was consistently more effective than the mouse MAb.
以SW948（一种人结肠癌细胞系）作为靶细胞，研究了健康个体外周血单个核细胞（PBMC）在抗体依赖的细胞介导的细胞毒性作用（ADCC）中的裂解能力。使用了三种单克隆抗体（MAb）：两种分别针对抗原结构CO17 - 1A和BR55 - 2的小鼠MAb（IgG2A）以及一种嵌合MAb 17 - 1A（IgG1）（鼠 - 人）。制备了三种效应细胞。PBMC在Ficoll - Isopaque梯度上纯化（FIP细胞）（淋巴细胞和单核细胞的混合物）。为获得纯度大于90%的纯单核细胞，PBMC在Nycodenz梯度上离心（Nycodenz细胞）。通过用铁粉处理FIP细胞并去除吞噬细胞获得高度纯化的淋巴细胞（大于98%）（PBL细胞）。单核细胞具有最高的裂解能力。FIP细胞的效果不如单核细胞。PBL细胞的杀伤活性最差。在重组实验中，向PBL中添加越来越多的单核细胞会导致细胞毒性增强。Leu - M3 +细胞、Leu - M5 +细胞（单核细胞）和CD16 +细胞的数量与细胞毒性呈正相关。与单核细胞相比，使用FIP细胞作为效应细胞时，需要更高浓度的MAb 17 - 1A才能达到相同水平的细胞毒性。MAb BR55 - 2诱导的细胞毒性活性与MAb 17 - 1A相同。这两种MAb的组合并未增加裂解能力。嵌合MAb 17 - 1A以剂量依赖方式介导ADCC。嵌合MAb始终比小鼠MAb更有效。
