自体LAK高效扩增法的建立及活性诱导实验  

HIGH EFFICIENCY AMPLIFICATION METHOD FOR AUTOLAK CELLS AND ACTIVITY INDUCING EXPERIMENT

作　　者：强新[1,2,3,4] 张顺[1,2,3,4] 于建宪 刘继民[1,2,3,4] 庞新建 宋修歧 

机构地区：[1]青岛医学院微生物学教研室 [2]青岛医学院附属医院普外科 [3]青岛市市立医院病理科 [4]青岛医学院第二附属医院外科

出　　处：《青岛医学院学报》1998年第3期160-162,共3页Acta Academiae Medicinae Qingdao Universitatis

基　　金：山东省卫生厅青年科学基金

摘　　要：目的建立体外高效扩增自体ＬＡＫ细胞方法，并测定诱导扩增的ＬＡＫ细胞是否具有杀瘤细胞作用及表型变化。②方法取２８例不同类型的肿瘤病人外周静脉血液，经密度梯度离心获取ＬＡＫ细胞前体，以白细胞介素２（ＩＬ－２）作为诱导剂，应用独特的液相三步扩增方法扩增自体ＬＡＫ细胞。分别用间接免疫荧光（ＩＦＡ）和四甲基偶氮唑盐（ＭＴＴ）法，测定细胞表型和对肿瘤细胞的杀伤活性，并与对照组细胞进行比较。③结果以该法诱导扩增４～１２ｄ，细胞数量增加１２０倍，对Ｋ５６２和Ｒａｊｉ细胞杀伤活性最高达６１．７％和４０．１％（效∶靶＝５０∶１），与对照组细胞比较，差异有极显著性（ｔ＝４．５～４．７，Ｐ＜０．０１）。自体ＬＡＫ细胞具有成熟Ｔ细胞表型特点（ＣＤ３和ＣＤ８）。④结论此法可扩增出高数量和高效杀肿瘤细胞活性的自体ＬＡＫ细胞。Objective To establish high effective amplification method for Auto-LAK cells and study the tumor cell killing activity and phenotype of the amplified auto-LAK cells Methods The peripheral blood was obtained from 28 patients with different tumors. LAK cells were separated by density gradient centrifugation and amplified with liquid threestep method in the presence of IL-2. The cell phenotype and killing activity were studied with IFA and MTT methods and compared with the control group. Results The LAK cells were increased to 120 folds after culture for 4-12d. The killing rates of LAK cell for K_562 and Raji cells were 61.7% and 40.1%(effect∶target=50∶1), which were significantly greater than those of the control group (t=4.5￣4.7,P<0.01). AutoLAK cell had phenotype of mature Tcells(CD3,CD8). Conclusion Liquid threestep amplification method could produce large amount of autoLAK cells which had great killing activity for tumor cells.

关 键 词：液相三步扩增法 肿瘤 LAK细胞 诱导实验 

分 类 号：R73－35[医药卫生—肿瘤]