小鼠脾内注射日本血吸虫病患者尿液循环抗原制备单克隆抗体

目的： 探讨用一种快速免疫方法制备抗人尿液日本血吸虫循环抗原的单克隆抗体。方法： 将日本血吸虫病患者尿液用１５％ 三氯醋酸提取循环抗原后采用脾内直接注射法免疫 Ｂ Ａ Ｌ Ｂ／ｃ小鼠制备单克隆抗体。结果： 建立了 ２ 株分泌抗人尿日本血吸虫循环抗原的单克隆细胞株２ Ｅ６ 及２ Ｂ１１，两者免疫球蛋白鉴定均属 Ｉｇ Ｍ 类， Ｅ Ｌ Ｉ Ｓ Ａ测得单抗对尿液循环抗原（ Ｉ Ｈ Ｕ Ｃ Ａｇ），效价分别为２．５６×１０５ 和６．４×１０４ 。琼脂双扩散试验显示２ 株单抗与正常人尿三氯醋酸提取物均不能形成沉淀线，而２ Ｅ６ 与日本血吸虫病患者尿液循环抗原能产生沉淀线。分别用２ Ｂ１１ 和２ Ｅ６ 腹水为捕捉抗体， Ｈ Ｒ Ｐ Ｈ１１ 为酶标抗体检测 １２ 例急性血吸虫病患者尿液，依次为６ 例和３ 例阳性，８ 例正常人尿液均为阴性。结论： 用血吸虫病患者尿液提取的循环抗原经脾脏注射免疫 Ｂ Ａ Ｌ Ｂ／ｃ 小鼠制备单克隆抗体用于检测日本血吸虫病患者尿液中循环抗原是可行的。

PREPARATION OF MONOCLONAL ANTIBODIES BY INTRASPLENIC IMMUNIZATION OF MICE WITH URINE CIRCULATING ANTIGEN FROM PATIENTS INFECTED WITH SCHISTOSOMA JAPONICUM

AIM: To explore an efficient method for preparing monoclonal antibodies against circulating antigen of S. japonicum in the urine of schistosomiasis patients. METHODS: The urine of schistosomasis patients were precipitated with 5% trichloroacetic acid, and then used to immunize the BALB/c mice by intrasplenic immunization to prepare McAb. RESULTS: Two monoclonal cell lines against the CAg in the urine of schistosomiasis patients have been established. The immunoglobulin subclass of the 2 McAbs (2E6 and 2B11) were identified as IgM. The titres of 2E6 and 2B11 were 2.56 x 10(5) and 6.4 x 10(4). On immunodiffusion, McAb 2E6 showed one precipitation line with IHU-CAg, however, both McAbs did not react with NHU-CAg. Using ascitic fluid of 2B11 and 2E6 as capture antibody separately, and HRP-H11 as labelled antibody to detect the urine of 12 acute schistosomasis patients, 2B11 2E6 gave 6 and 3 positive, respectively, whereas 8 normal urine were all negative. CONCLUSION: It is feasible to use CAg from schistosomasis patients' urine to prepare anti-CAg McAb by intrasplenic immunization in mice.