Karpovsky B, Titus J A, Stephany D A, Segal D M
J Exp Med. 1984 Dec 1;160(6):1686-701. doi: 10.1084/jem.160.6.1686.
Rabbit anti-2,4-dintrophenyl (DNP) antibodies or their F(ab')2 fragments were chemically cross-linked to the anti-mouse Fc gamma R monoclonal antibody 2.4G2 or to its Fab fragment. P388D1 cells were incubated with heteroaggregates between 2.4G2 and anti-DNP (anti-Fc gamma R X anti-DNP) and washed. The resulting cells lysed 2,4,6-trinitrophenyl chicken erythrocytes (TNP CRBC) in a hapten-specific manner. The lysis was inhibited by free hapten but was resistant to inhibition by immune complexes. Other cells coated with antibody heteroaggregates also mediated lysis of TNP-modified target cells. For example, mouse resident peritoneal exudate cells (PEC) lysed TNP CRBC and bacillus Calmette-Guérin-activated PEC lysed both TNP CRBC and TNP tumor targets. Human neutrophils, when incubated with heteroaggregates containing the anti-human neutrophil Fc gamma R antibody 3G8 and anti-DNP also lysed TNP CRBC and TNP-modified tumor cells. To test whether linkage to Fc gamma R was required for lysis, F(ab')2 fragments from the anti-KdDd monoclonal antibody 34-1-2 were cross-linked to anti-DNP F(ab')2 fragments. P388D1 cells (which express Kd and Dd) were then incubated with these heteroaggregates and washed, and their abilities to form conjugates and lyse TNP CRBC were compared with those of P388D1 cells treated with anti-Fc gamma R X anti-DNP. In both cases, P388D1 cells formed conjugates. However, only the cells treated with anti-Fc gamma R X anti-DNP mediated lysis to a significant extent. We conclude that heteroaggregates containing anti-Fc gamma R and anti-target cell antibodies can be used to create potent effector cells against red cell and tumor targets and that bridging of effectors with target cells directly to Fc gamma R on effector cells is required for lysis.
兔抗2,4 - 二硝基苯基（DNP）抗体或其F(ab')2片段与抗小鼠FcγR单克隆抗体2.4G2或其Fab片段进行化学交联。将P388D1细胞与2.4G2和抗DNP之间的异聚体（抗FcγR×抗DNP）一起孵育并洗涤。所得细胞以半抗原特异性方式裂解2,4,6 - 三硝基苯基鸡红细胞（TNP CRBC）。这种裂解被游离半抗原抑制，但对免疫复合物的抑制具有抗性。其他包被有抗体异聚体的细胞也介导TNP修饰靶细胞的裂解。例如，小鼠驻留腹膜渗出细胞（PEC）裂解TNP CRBC，卡介苗激活的PEC裂解TNP CRBC和TNP肿瘤靶细胞。人中性粒细胞与含有抗人中性粒细胞FcγR抗体3G8和抗DNP的异聚体一起孵育时，也裂解TNP CRBC和TNP修饰的肿瘤细胞。为了测试裂解是否需要与FcγR连接，将抗KdDd单克隆抗体34 - 1 - 2的F(ab')2片段与抗DNP F(ab')2片段交联。然后将P388D1细胞（表达Kd和Dd）与这些异聚体一起孵育并洗涤，并将它们形成结合物和裂解TNP CRBC的能力与用抗FcγR×抗DNP处理的P388D1细胞进行比较。在两种情况下，P388D1细胞都形成了结合物。然而，只有用抗FcγR×抗DNP处理的细胞在很大程度上介导了裂解。我们得出结论，含有抗FcγR和抗靶细胞抗体的异聚体可用于产生针对红细胞和肿瘤靶标的强效效应细胞，并且效应细胞与靶细胞直接桥接至效应细胞上的FcγR对于裂解是必需的。
