摘要
目的 研究杀伤细胞对人白血病多药耐药（ＭＤＲ） 细胞系的杀伤作用。方法 （１） 采用体外药敏试验（ＭＴＴ法） 检测两株ＭＤＲ 细胞系，即Ｋ５６２／ＶＣＲ和Ｋ５６２／ＨＨＴ 细胞的耐药性。（２） 用优选方案诱导效应细胞：５０ ｎｇ／ｍｌ 的抗ＣＤ３ 单克隆抗体（ＣＤ３ＭｃＡｂ） 和５０ Ｕ／ｍｌ 的ＩＬ２ 诱导的ＣＤ３ＡＫ 细胞（ＣＤ３ＭｃＡｂａｃｔｉｖａｔｅｄ ｋｉｌｌｅｒｃｅｌｌ）；４００ Ｕ／ｍｌ 的ＩＬ２ 诱导淋巴因子激活的杀伤细胞（ｌｙｍｐｈｏｋｉｎｅ ａｃｔｉｖａｔｅｄ ｋｉｌｌｅｒｃｅｌｌ，ＬＡＫ） 。（３） 以３ＨＴｄＲ掺入法测定效应细胞的增殖活性。（４） 以３ＨＴｄＲ 释放法测定效应细胞对Ｋ５６２ 细胞及其两个ＭＤＲ细胞系的杀伤活性。结果 （１） Ｋ５６２／ＶＣＲ、Ｋ５６２／ＨＨＴ经冻存复苏后无药培养１～２ 个月仍具有高度耐药性。（２） 成功地诱导出ＣＤ３ＡＫ＋／ － 细胞及ＬＡＫ 细胞。（３）ＣＤ３ＡＫ＋ 细胞的增殖活性较ＬＡＫ细胞强；ＣＤ３ＡＫ＋ 细胞对Ｋ５６２ 细胞的杀伤活性较ＬＡＫ 细胞强，且维持时间较ＬＡＫ 细胞长。（４）ＣＤ３ＡＫ＋ 细胞对Ｋ５６２／ＶＣＲ 细胞的杀伤活性强于对敏感细胞Ｋ５６２，其对Ｋ５６２?
Objective To study the cytotoxicities of the killer cells to human multidrug resistant (MDR) leukemic cell lines Methods (1) The sensitivities of two MDR cell lines (K562/VCR and K562/HHT) reacted to vincristine (VCR), high harringtonine (HHT) and daunorubicin (DNR) were examined by MTT assay (2) For induction of effector cells, 50 ng/ml of CD 3 monoclonal antibody (CD 3 McAb) and 50 U/ml of IL 2 were selected as the optimal condition for the induction of CD 3McAb activated killer cells (CD 3AK cells); 400 U/ml of IL 2 was used for the generation of lymphokine activated killer cells (LAK cells) (3)Proliferations of the effector cells were determined by 3H TdR uptake assay (4) Cytotoxicities of the killer cells to K562 as well as the two MDR cell lines of K562 were determined by 3H thymidine ( 3H TdR) release assay Results (1) The multi drug resistance consistently existed in both cell lines of K562/VCR and K562/HHT thawed after freezing and cultured for 1～2 months with drug free medium (2) CD 3AK +/- and LAK cells were successfully induced (3) The proliferation of CD 3AK + cells was much stronger than that of LAK cells The cytolysis of K562 cells by CD 3AK + cells was higher than that of CD 3AK - and LAK cells The duration of the cytolytic activity of CD 3AK + cells was longer than that of the LAK cells (4) The hilling activing of CD 3AK + cells showed significantly higher to K562/VCR cells than to K562 cells, while there was no difference of cytotoxicity between K562 /HHT and K562 cells No significant difference was found in LAK mediated cell killing either to K562/VCR and K562 /HHT cells or to K562 cells The cell mediated killing of K562/VCR cells by CD 3AK + cells was significantly higher than that by LAK cells Conclusion CD 3AK + cells might be used as effector cells in adoptive immunotherapy of drug resistant leukemic patients
出处
《中华儿科杂志》
CSCD
北大核心
1999年第9期543-546,共4页
Chinese Journal of Pediatrics
基金
浙江省自然科学基金
关键词
白血病
单克隆抗体
杀伤细胞
多药抗药性
Leukemia
Antibodies
monoclonal
Killer cells
Drug resistance
multiple
