抗弓形虫M cA b的建立及应用研究

ESTABLISHMENT AND APPLICATION OF ANTI-TOXOPLASMA GONDII McAbs

应用杂交瘤技术 ,建立了 5株分泌抗弓形虫单克隆抗体的杂交瘤细胞。其中 1E9和 4C10 2株分泌的 Mc Ab效价达 1∶ 12 80 0。该 2株单克隆抗体均属 Ig G1亚类 ,其识别的弓形虫抗原分别为 P2 2、P2 8。1E9和 4C10 Mc Ab混合后与兔抗弓形虫抗体联合 ,进行双抗体夹心 EL ISA测定弓形虫的敏感性分别为 12 .5和 2 5个虫体 / m l。检测 146份有不良孕产史妇女血清 ,11份弓形虫循环抗原 ( CAg)阳性 ( 7.5 3% )。检测 11份自然流产物 ,1份 CAg阳性 ,该流产物转种小鼠后证实确含有弓形虫。 92份育龄妇女血清 1份弓形虫 CAg阳性 ( 1.0 9% )。有不良孕产史的妇女血清弓形虫 CAg阳性率明显高于育龄妇女血清阳性率 ( P<0 .0 5 )。弓形虫经 1E9、4C10 Mc Ab处理后对小鼠腹腔巨噬细胞攻击 ,感染率分别为 7.8%和 7.5 % ,而对照组感染率为 14.5 % ,证明该

strains of hybridoma secreting monoclonal antibody against Toxoplasma gondii were established. Among the five secreted monoclonal antibodies (McAbs), McAb 1E9 and McAb 4C10 were found to be able to recognize P22 and P28 T. gondii antigens respectively. Both of them belong to IgG1. The mixture of McAb 1E9 and McAb 4C10, and rabbit antiserum against T. gondii were used in sandwich ELISA to detect specific circulating antigens. 11 of the 146 patients with abnormal pregnancy or abnormal childbearing history were positive (7.53%) and 1 of 11 samples of natural apoblema was positive, which was confirmed by mouse inoculation. 1 of 92 blood samples collected from women of childbearing age was positive (1.09%). The sensitivity of sandwich ELISA for detecting T. gondii was 12.5 parasites/ml (1E9) and 25 parasites/ml (4C10). The fact that the positive rates of mouse peritoneal macrophage (7.8%, 7.5%) challenged with McAb 1E9 or McAb 4C10treated T. gondii were lower than with untreated parasite (14.5%). It is indicated that both McAbs had inhibitory effect on T. gondii.