健康人和肿瘤患者CIK细胞的生物学特性比较及应用

The comparison and application of the biological characteristics of CIK cells from healthy donors and tumor patients

目的比较健康人和肿瘤患者外周血来源的CIK细胞的体外扩增速度、细胞表型和杀伤活性;观察健康人CIK细胞对裸鼠体内移植瘤的预防和治疗作用。方法健康人和肿瘤患者外周血单个核细胞各10份,定向诱导成CIK细胞,直接活细胞计数法比较两者的扩增速度,流式细胞仪检测比较两者细胞表型,MTT法比较两者对K562和LOVO细胞株的杀伤活性。取30只裸鼠分为3个组,预防组:先尾静脉注射CIK细胞,后背部皮下接种A549细胞。治疗组:裸鼠于第1天接种A549细胞,第2天,根据注射方案不同,分为局部注射CIK细胞(局部注射组)、尾静脉注射CIK细胞(尾静脉注射组)、局部及尾静脉均注射CIK细胞(联合治疗组)3组,均连续治疗5 d。对照组:背部皮下接种A549细胞,注射生理盐水,连续5 d。4周后裸鼠,测量肿瘤大小,称重,计算肿瘤体积、抑瘤率,并做病理检查。结果健康人CIK细胞的扩增速度显著高于肿瘤患者CIK细胞(P<0.05),流式细胞仪检测显示健康人CIK细胞CD3+CD8+、CD3+CD56+细胞百分比显著高于患者CIK细胞(P<0.05),健康人CIK细胞对K562、LOVO的杀伤活性强于患者CIK细胞(P<0.05)。对照组各裸鼠背部皮下移植瘤出现较早,并且以相近速率快速生长,体积较大;CIK细胞预防组及治疗组中局部注射组和尾静脉注射组的移植瘤出现较晚,生长较慢,体积较小,联合治疗组治疗效果更佳(P<0.05)。结论体外实验中,健康人CIK细胞体外扩增快,CD3+CD8+、CD3+CD56+细胞比例高,对肿瘤细胞的杀伤活性强于肿瘤患者CIK细胞。动物实验中,健康人CIK细胞对裸鼠移植瘤有预防和治疗作用。

Objective To compare the proliferation, phenotypes and cytotoxicity of CIK cells between healthy donors and tumor patients, and to evaluate the preventive and curative effects of CIK cells on transplanted tumor cells. Methods Each 10 doses of peripheral mononuelear cells from healthy donors and tumor patients were collected and sub- sequently induced into CIK cells. The proliferation activity and phenotypes of CIK cells were assessed by cell count and flow eytometry, respectively. The cytotoxie activities to K562 cells and LOVO cells of CIK ceils were determined by MTr method. Thirty nude mice were randomly divided into prevention, treatment, and control groups. In prevention group, everyday caudal injection of CIK cells was performed for 5 days. On the 6th day, subcutaneous injection of A549 cells on the back was carried. In the treatment group, A549 cells were injected on back subcutaneously in 18 nude mice on the 1st day, which were randomly assigned into subgroups with different treatment from the 2nd day. In the control group, CIK cell injection was substituted by saline as control. All subjects were sacrificed at week 4th for assessment of tumor size, weighing, tumor volume, inhibitory rate, and pathological changes. Results The proliferation velocity of CIK cells was significantly higher from healthy donors than that from tumor patients （ P 〈 0. 05 ）, so were the CD3 ＋ CD8 ＋ and CD3 ＋ CD56 ＋ positive rates of CIK cells （ P 〈 0. 05 ）, and the anti - tumor effect on K562 and LOVO cell line of CIK cells （ P 〈 0. 05）. Early emergence, rapid growth and the large volume of transplanted tumor in control group was observed; while significantly later emergence, slower growth and smaller volume were characterized in prevention and treatment group, with most prominent effect of combination treatment subgroup （ P 〈 0. 05 ）. Conclusion CIK with higher proliferation velocity, CD3 ＋ CD8 ＋ and CD3 ＋ CD56 ＋ positive rates, and cytotoxieity is observed from of healthy donors than that form tumor patients in vitro. CIK cells from healthy donors show preventive and therapeutic effect on transplant tumor in nude mice.