CHARACTERIZATION OF A NEW MONOCLONAL ANTI-FC-GAMMA-RII ANTIBODY, AT10, AND ITS INCORPORATION INTO A BISPECIFIC F(AB')2 DERIVATIVE FOR RECRUITMENT OF CYTOTOXIC EFFECTORS
CHARACTERIZATION OF A NEW MONOCLONAL ANTI-FC-GAMMA-RII ANTIBODY, AT10, AND ITS INCORPORATION INTO A BISPECIFIC F(AB')2 DERIVATIVE FOR RECRUITMENT OF CYTOTOXIC EFFECTORS
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DOI：
10.1016/0161-5890(91)90011-8
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发表时间：
1991-11-01
影响因子：
3.6
通讯作者：
GLENNIE, MJ
中科院分区：
文献类型：
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作者：
GREENMAN, J;TUTT, AL;GLENNIE, MJ
单核细胞上的Fc-γ-RII（CDw 32）能够触发包被有适当同种型抗体的鸡红细胞（CRBC）的吞噬作用和裂解作用。在这份报告中，我们描述了生产和表征的小鼠单克隆IgG 1抗体特异性Fc-γ RII和比较其在结合研究，组织分布和重定向细胞毒性（RCC）的活性，与以前确定的抗Fc-γ-RII抗体KB 61和IV.3。免疫组织化学和流式细胞术分析表明，AT 10结合非常强烈的Fc-γ RII正常单核细胞，但只有弱的淋巴细胞上表达。该模式不对应于用KB 61或IV. 3观察到的染色，并且似乎给出中间特征。AT 10的Fab'片段的结合常数（Ka）计算为5.3 × 10（8）M-1，比KB 61的结合常数（1.4 × 10（8）M-1）高4倍。从AT 10或KB 61的Fab'片段与来自抗CRBC单克隆抗体的Fab'硫醚连接构建双特异性F（ab'）2抗体。这些双特异性衍生物与单克隆或多克隆抗鸡红细胞抗体一样有效地指导单核细胞对CRBC的细胞毒性。双特异性F（ab '）2抗体具有优于常规试剂的明显优势，因为它们在3.5mg/ml（与血清中IgG提供的浓度相当的浓度）的人Fc-γ存在下不被阻断。因此，用高亲和力抗Fc-γ RII抗体AT 10构建的双特异性衍生物可用作体内靶向肿瘤细胞裂解的治疗试剂。
Fc-gamma-RII (CDw32) on monocytes is capable of triggering both phagocytosis and lysis of chick red blood cells (CRBC) coated with antibody of the appropriate isotype. In this report we describe the production and characterization of a mouse monoclonal IgG1 antibody specific for Fc-gamma RII and compare its activity in binding studies, tissue distribution and redirected cellular cytotoxicity (RCC), with the previously identified anti-Fc-gamma-RII antibodies KB61 and IV.3. Immunohistochemical and flow cytometry analyses demonstrated that AT10 binds very strongly to Fc-gamma RII on normal monocytes, but only weakly to that expressed on lymphocytes. This pattern does not correspond to the staining seen with either KB61 or IV.3, and appears to give an intermediate profile. The binding constant (Ka) for the Fab' fragment of AT10 was calculated at 5.3 x 10(8) M-1, four times higher than that for KB61 (1.4 x 10(8) M-1).Bispecific F(ab')2 antibodies were constructed from Fab' fragments of AT10 or KB61 thioether-linked to Fab' from an anti-CRBC monoclonal antibody. These bispecific derivatives directed monocyte cytotoxicity against CRBC as efficiently as either a monoclonal or polyclonal anti-chick erythrocyte antibody. The bispecific F(ab')2 antibodies had a distinct advantage over the conventional reagents, in that they were not blocked in the presence of human Fc-gamma at 3.5 mg/ml (a concentration comparable with that provided by IgG in serum). Therefore, bispecific derivatives constructed with the high affinity anti-Fc-gamma RII antibody, AT10, may be used as therapeutic reagents for targeting tumour cell lysis in vivo.