用Epstein-Barr病毒转化技术建立分泌人单克隆抗体B淋巴细胞系的条件  

CONDITIONS FOR ESTABLISHMENT OF HUMAN MONOCLONAL ANTIBODY PRODUCING B LYMPHOBLASTOID CELL LINES USING EPSTEIN-BARR VIRUS

作　　者：房强[1] 郁文芳 陈伯权[1] 刘琴芝[1] 宋光运 陶三菊[1] 杨冬荣[1] 

机构地区：[1]中国预防医学科学院病毒学研究所

出　　处：《病毒学报》1992年第2期179-183,共5页Chinese Journal of Virology

摘　　要：用以去除T淋巴细胞的E—玫瑰花形成法中所用的羊红细胞(SRBC),其保鲜程度对EB病毒转化B淋巴细胞的结果有一定影响,用保存1天的SRBC者,转化细胞的抗体分泌时间较长(8～10周),并且能形成传代细胞系;用保存17天的SRBC,转化细胞的抗体分泌时间较短,未能无限传代。分离外周血单个核细胞时,保留自体粘附细胞作为饲养细胞,转化的细胞二个月的成活率可达75％,而不保留粘附细胞者,转化细胞成活率仅40％。能产生抗-HBc抗体的EBV转化细胞,自然分泌抗体时间一般不超过10周,高峰期在3周以内,第2周时最高。EB病毒感染后的转化细胞5天开始克隆,一般细胞难成活,在14天克隆的,成活率可达100％。21天开始克隆的转化细胞,免疫球蛋白自然分泌维持12周以上,而在35天开始克隆的细胞,维持分泌的时间都不超过10周。以未经γ线照射的小鼠腹腔渗出细胞作饲养细胞,可支持转化3周的细胞克隆扩增,克隆成活率可达100％,而对照组克隆成活率仅为64.7％。We separated human T cells from non-T cells by E-rosette test.When sheep erythrocytes ( SRBC)stored at 4℃ for one day was used in the test, EBV-transformed cells maintained anti-HBc secretion for 8-10 weeks and an immortalized rate of 100%. In another group in which SRBC stored at 4℃ for 17 days was used, EBV-transformed cells secreted anti-HBc for only 6 week and there was no immortalization. EBV-transformed cells with adhesive mononuclear cells as feeder survived 75% for two months, while the survival rate of the control without the feeder cells wat 40%. Anti-HBc antibody-producing transformed cells maintained Ig secretion for less than 10 weeks and the highest level was within 3 weeks. Maintenance of Ig secretion of EBV-transformed cells cloned at 21 days after EBV infection was above 12 weeks,but those cloned at day 35,less than 10 weeks. Peritoneal exudate cells(PC)from 6-week-old swiss mice were used as feeder cells to support EBV-transformed cell cloning at day 21 post EBV infection. The group with PC had higher survival rate than the control without PC ( 100% vs 64.7%) . Additionally, EBV-transformed cells survived not more than 12.5% when clonied on day 5 and 100% when cloned on day 14.

关 键 词：E-B病毒 转化细胞系 人单克隆抗体 

分 类 号：R392－33[医药卫生—免疫学]