BISPECIFIC F(AB')2 MONOMER PREPARED WITH ANTI-CD3 AND ANTI-TUMOR MONOCLONAL-ANTIBODIES IS MOST POTENT IN INDUCTION OF CYTOLYSIS OF HUMAN T-CELLS
BISPECIFIC F(AB')2 MONOMER PREPARED WITH ANTI-CD3 AND ANTI-TUMOR MONOCLONAL-ANTIBODIES IS MOST POTENT IN INDUCTION OF CYTOLYSIS OF HUMAN T-CELLS
复制标题
DOI：
10.1002/eji.1830190814
复制
发表时间：
1989-08-01
影响因子：
5.4
通讯作者：
OKUMURA, K
中科院分区：
文献类型：
--
作者：
NITTA, T;YAGITA, H;OKUMURA, K
制备由抗CD3单抗和抗肿瘤单抗Fab‘’片段组成的双特异性F(ab‘)2，研究其诱导外周血单核细胞对靶细胞的杀伤活性。在用二硫苏糖醇还原这些片段的交换二硫键后，加入硫醇-二硫键链间试剂5，5‘-二硫双-2-硝基苯甲酸，将其中一个Fab’‘片段的自由SH基团转化为混合的二硫代衍生物。然后将这些片段与其他含有游离SH基团的Fab‘’片段偶联，生成高产率的双特异性杂交物F(ab‘’)2单体。双特异性F(ab‘)2单体能使未激活的外周血单核细胞对自然杀伤细胞耐药肿瘤细胞株产生1微克/毫升的细胞毒作用，用交联剂N-succinimidyl-3-(2-pyridyldithiol)-propionate(SPDP)或S-乙酰硫代琥珀酸酐(SAMSA)制备的F(ab’)2片段的聚合体诱导细胞杀伤活性低于单体F(ab‘)2片段。双特异性F(ab‘)2单体由于制备简单，而且由于其小尺寸(100-110 kDa)而在诱导细胞杀伤活性和高组织通透性方面具有很高的效率，因此将其用于癌症免疫治疗是可行的。此外，网状内皮系统细胞通过Fc-Fc受体相互作用将这两种类型的细胞连接起来而导致的外周血单核细胞重定向细胞溶解活性没有发生。
Induction of cytolytic activity of peripheral blood mononuclear cells towards target cells was studied by preparing bispecific F(ab'')2 which was composed of two Fab'' fragments, one of which was derived from anti-CD3 monoclonal antibody and the other from anti-tumor monoclonal antibody. After reduction of the interchange disulfide bonds of these fragments by dithiothreitol, a thiol-disulfide interchain reagent, 5,5''-dithiobis-2-nitrobenzoic acid, was added to convert the free SH groups of one of the Fab'' fragments to mixed disulfide derivatives. These were then coupled with the other Fab'' fragment bearing free SH groups, producing a bispecific hybird F(ab'')2 monomer of high yield. The bispecific F(ab'')2 monomers were able to render nonactivated periphral blood mononuclear cells cytotoxic against natural killer-resistant tumor cell lines at doses as low as 1 .mu.g/ml. The polymeric forms of the F(ab'')2 fragments prepared by use of cross-linking reagents such as N-succinimidyl-3-(2-pyridyldithiol)-propionate (SPDP) or S-acetylmercaptosuccinic acid anhydride (SAMSA) were less efficient for induction of cytolytic activity than the monomeric one. It may be feasible to use bispecific F(ab'')2 monomers in cancer immunotherapy because of the ease of preparation, as well as the efficiency in inducing cytolytic activity and their high tissue permeability due to their small size (100-110 kDa). In addition, redirected cytolytic activity towards peripheral blood mononuclear cells by reticuloendothelial system cells, resulting from linking these two types of cells through Fc-Fc receptor interactions, does not occur.