免疫删减法制备共刺激分子CD80单克隆抗体及其生物学活性鉴定  

The Production of monoclonal antibody to CD80 by subtrative immunizationtechniques and identification of its biological activity

作　　者：周璟[1] 马宝骊[1] 严俊[1] 张继英[1] 张汉明[1] 

机构地区：[1]上海第二医科大学免疫学教研室,上海200025

出　　处：《中国免疫学杂志》2000年第1期35-38,共4页Chinese Journal of Immunology

摘　　要：目的：制备共刺激分子ＣＤ８０单抗并鉴定其生物学活性。方法；采用给经ＭＤＡ４５３细胞免疫的小鼠注射环磷酰胺１００ｍｇ／（ｋｇ·次）诱导对ＭＤＡ４５３细胞的免疫耐受基础上，再用已建立能稳定表达ｈＣＤ８０分子的ＭＤＡ４５３细胞免疫小鼠，取经检测抗体阳性强的小鼠脾细胞，按常规方法制备单抗。通过Ｔ增殖阻断实验鉴定其活性。结果：ＣＥＬＬ－ＥＬＩＳＡ、ＦＡＣＳ鉴定获得一株抗ＣＤ８０单抗，腹水效价为１０－６，属ＩｇＧ１亚类。Ｔ细胞增殖阻断试验显示，此单抗能部分阻断Ｔ细胞对相应刺激的增殖反应。结论：应用环磷酰胺免疫删减法成功制备膜抗原ＣＤ８０单抗，该单抗有一定应用前景。Objective: Preparing CD80 McAb and identifying its biological activity. Methods: Mice were exposed to the MDA453 cell(tolerogen ) followed at 15 min,24 h, 48 h by injection of cyclophosphamide(CY). The MDA453 tolerized mice then immunized with stable hCD80 trandfectant MDA453, cell. Results: One stable hybridoma cell line secreting McA-b anti-CDSO has been established. The specificity ofMcAb were identified by cell-ELISA and FACS. The McAb belongs to IgG1 subclass. The bier of McAb in ascites was 10-6. The McAb couldpartly inhabit T lymphocyte proliferation to the proper stimulations. Conclusion: Subtractive immunization techniques can be employed to produce CD80 McAb. The McAb has a potential use.

关 键 词：环磷酰胺 免疫耐受 单克隆抗体 CD80 

分 类 号：R392－33[医药卫生—免疫学]