Pathological and biochemical significance of the complex between kininogens and thiol proteinases

制备抗高分子量激肽原-钙蛋白酶I复合体(CL复合体)的单抗。获得的5株单抗分别命名为肝癌-1、2、3、4和5，可与CL复合体反应，但不与其新生成分、高分子激肽原或钙蛋白酶I反应。其免疫球蛋白类型为IgM，轻链型为Kappa。只有HCC-3和-4能识别非交联型高分子量激肽原-钙蛋白酶I复合体(N复合体)，而另一种不能识别。肝细胞癌-3对N复合体的识别能力强于肝细胞癌-4。HCC-3还识别其他N复合体：高分子激肽原-钙蛋白酶II复合体、低分子激肽原-钙蛋白酶I复合体和低分子激肽原-钙蛋白酶II复合体。Hcc-3识别的表位可能是高分子量激肽原与calain I相互作用形成的特殊结构，以及四种激肽原-calain复合体共有的结构。用夹心ELISA法测定肝癌细胞的CL复合体含量可达1 ng/ml，N复合体含量可达100 ng/ml。正常血浆(n=17)中N-复合体水平<100 ng/ml，而肝细胞癌(MAX)患者10份血浆中有4份可检测到N-复合体。426 ng/ml)，3例中有2例为肝硬变(最大311 ng/ml)，1例肝母细胞瘤(290 ng/ml)。这是首次报道在体内存在与激肽原和钙蛋白酶之间的复合物的单抗。

Monoclonal antibodies were raised against cross-linked high molecular weight kininogen- calpain I complex (CL complex). Five monoclonal antibodies obtained were designated as HCC- 1, 2, 3, 4 and 5, and found to react with CL complex, but not with its nascent components, high molecular weight kininogen or calpain I either. Their immunoglobulin class and light chain type are IgM and kappa, respectively. Only HCC-3 and -4 recognized non cross-linked high molecular weight kininogen-calpain I complex (N complex), but the other did not. HCC-3 recognized the N complex stronger than HCC-4. HCC-3 also recognized other N complexes, high molecular weight kininogen-calpain II complex, low molecular weight kininogen-calpain I complex and low molecular weight kininogen-calpain II complex. The epitope recognized by HCC-3 would be the specific structure formed by the interaction between high molecular weight kininogen and calpain I, and its structure common to four kininogen-calpain complexes. Employing sandwich ELISA assay system with HCC-3, it was possible to estimate the amount of the complex up to 1 ng/ml for CL complex and 100 ng/ml for N complex. The level of N complex in normal plasma (n=17) was less than 100 ng/ml. However, we could detect the complex in 4 of 10 plasma samples from the patients with hepatocellular carcinoma (Max. 426 ng/ml), 2 of 3 with liver cirrhosis (max. 311 ng/ml), and 1 of 1 with hepatoblastoma (290 ng/ml).This is the first report which indicated the presence of the complex in vivo with the monoclonal antibody against a complex between kininogen and calpain.