家兔对小鼠抗血吸虫靶抗原单克隆抗体的抗抗体应答

本文报告从分泌具血吸虫保护性IgG_(2a)表型1E2McAb的杂交瘤培养上清液中,应用蛋白A-葡聚糖柱层析法纯化的McAb免疫家兔,采用纯化McAb分别进行Dot-ELISA、免疫双扩散和免疫印迹试验、ELISA观察免疫血清中抗McAb和抗-抗McAb的产生及其动态变化规律。主要结果表明:(1)1E2McAb免疫后,20d开始出现抗McAb,若不加强免疫,抗McAb逐渐减弱:若加强免疫,则抗McAb水平较高,且持续30-50d。(2)免疫血清中的抗-抗McAb,能起抗-抗Id的作用,可识别不同的血吸虫抗原及血吸虫抗原中不同抗原决定簇(90,68和45kD_a),而且免疫后不同时期抗-抗McAb识别不同抗原的水平不同(滴度1:100-1:1600)。对抗血吸虫肥抗原McAb免疫后抗抗体应答的观察,可为研制不同抗独特型McAb创造条件。

ANTI-ANTIBODY RESPONSES OF RABBIT TO MOUSE MONOCLONAL ANTIBODY AGAINST SCHISTOSOME "TARGET ANTIGEN"

The supernatant of protective McAb (isotype IgG2a) secreted by 1E2 hybridoma line was purified by protein A-sepharose CL-4B column and rabbits were immunized with purified 1E2 McAb. Dot-ELISA, immunodiffusion assay, Western blotting and ELISA were used to observe the presence of anti-McAb and anti-anti-McAb. The main results are: 1. anti-McAb antibody could be detected as early as 20 days post immunization in the immune sera and would be decreased gradually without boosting; otherwise, it would be increased quickly and the liters of anti-McAb were increased within 5 days postboosting and could be maintained at a plateau for 30-50 days, and then decreased gradually to the normal; 2. there were different anti-anti-McAbs in the immune sera. Anti-anti-McAb, like anti-anti-idiotype antibody, can recognize different schistosome antigens and 90, 68 or 45 kDa antige.nic determinants. The level of anti-anti-McAbs varied from tilers 1:100 to 1:1 600 at different time post-immunization with 1E2. The results provide evidence to understand the appearance of anti-McAb antibody and the way to develop anti-idiotype and anti-anti-idiotype McAb for research purpose.