CAJ | 학술논문

目的探讨源自健康人和肿瘤患者的细胞因子诱导的杀伤细胞(CIK)体外增殖能力及对原代肝癌细胞的抗肿瘤作用的差异,进一步了解CIK的抗肿瘤作用.方法分离健康人和肝癌患者外周血单个核细胞经细胞因子激活诱导培养为CIK,流式细胞分析检测CIK免疫表型;用机械研磨法将新鲜肝癌组织标本分离成单细胞悬液;四甲基偶氮唑盐法榆测两种CIK对原代肝癌细胞的杀伤活性.结果健康人和肝癌患者的CIK中CD3+ CD56+双阳性细胞数量均随培养时间的延长而增加,健康人CD3+ CD56+双阳性细胞所占比例在细胞因子诱导的第1、7、14天分别为1.053%±0.22%、25.36%±2.19%和55.12%±1.99%,均高于肝癌患者.重复测量方差分析显示健康人CIK体外扩增能力高于肝癌患者(P＜0.05),第1、7、14天3个时间点CD3+ CD56+ 双阳性细胞所占比例差异有统计学意义(P＜0.05).健康人和肝癌患者CIK对原代肝癌细胞的细胞毒活性与效靶比、作用时间有关.CIK与原代肝癌细胞作用24 h后,在10∶1、20∶1和40∶1等3个效靶比条件下,健康人CIK的细胞毒活性在不断增加,并且均高于肝癌患者(P＜0.05);CIK与原代肝癌细胞作用48 h小时后,在3个效靶比条件下,健康人CIK的细胞毒活性依次升高,并均高于肝癌患者(P＜0.05).结论源自健康人和肝癌患者的CIK对原代肝癌细胞都有细胞毒作用,但健康人CIK的增殖能力和细胞毒活性均高于肿瘤患者,因此,健康人来源的CIK可能具有更强的肿瘤杀伤作用.
목적탐토원자건강인화종류환자적세포인자유도적살상세포(CIK)체외증식능력급대원대간암세포적항종류작용적차이,진일보료해CIK적항종류작용.방법분리건강인화간암환자외주혈단개핵세포경세포인자격활유도배양위CIK,류식세포분석검측CIK면역표형;용궤계연마법장신선간암조직표본분리성단세포현액;사갑기우담서염법유측량충CIK대원대간암세포적살상활성.결과건강인화간암환자적CIK중CD3+ CD56+쌍양성세포수량균수배양시간적연장이증가,건강인CD3+ CD56+쌍양성세포소점비례재세포인자유도적제1、7、14천분별위1.053%±0.22%、25.36%±2.19%화55.12%±1.99%,균고우간암환자.중복측량방차분석현시건강인CIK체외확증능력고우간암환자(P＜0.05),제1、7、14천3개시간점CD3+ CD56+ 쌍양성세포소점비례차이유통계학의의(P＜0.05).건강인화간암환자CIK대원대간암세포적세포독활성여효파비、작용시간유관.CIK여원대간암세포작용24 h후,재10∶1、20∶1화40∶1등3개효파비조건하,건강인CIK적세포독활성재불단증가,병차균고우간암환자(P＜0.05);CIK여원대간암세포작용48 h소시후,재3개효파비조건하,건강 인CIK적세포독활성의차승고,병균고우간암환자(P＜0.05).결론원자건강인화간암환자적CIK대원대간암세포도유세포독작용,단건강인CIK적증식능력화세포독활성균고우종류환자,인차,건강인래원적CIK가능구유경강적종류살상작용.
Objective To explore the differences of the proliferation ability and the antineoplastic activity of CIK cells against origenal hepatocellullar cancer cells between healthy adults and tumor patients in vitro. Methods Peripheral blood mononuclear cells (PBMC) from healthy donors and tumor patients were incubated to induce CIK cells in the presence of interferon gamroa(IFN-γ) ,IL-2 and anti-CD3 monoclonal antibody (mAb). The changes in the proliferation activity and phenotypes of the CIK cells were identified by flow cytometric analysis. Single cell suspension was prepared bom the fresh hepatocellular carcinoma tissue by using mechanical trituration method. MTT assays were used to determine the cytotoxicity of CIK cells against origenal hepatocellullar cancer cells. Results The CIK cells from both healthy donors and tumor patients were significantly increased with the extension of time. The expression rate of CD3+/CD56+ cells from healthy donors rose from 1.053% ±0.22% on 1st day ,25.36% ±2.19% on the 7th day to 55.12% ±1.99% on the 14th day ,which was significantly higher than that of tumor patients ( P ＜ 0.05). MTT assays showed that the cytotoxicity of CIK cells from both enhanced obviously with the addition of Effect/Target rate and extension of time(P ＜0.05). After the origenal hepatocellullar cancer cells were treated by CIK cells 24 hours, the cytotoxicity of CIK cells from healthy donors at the effector:target ratio of 10 ∶ 1,20 ∶1 and 40 ∶1 was significantly higher than that of tumor patients(P＜0.05) ,respectively. After treated 48 hours, compared with tumor patients, the cytotoxicity of healthy donors? CIK at the three effector:target ratio was alsosignificantly higher(P ＜0.05), respectively. Conclusion CIK cells from both have amplification ability and cytotoxic activity in vitro,and the proliferation ability and killing activity of healthy adults CIK is stronger than that of tumor patients,which provides an experimental basis for CIK to clinical application as an adoptive immunotherapy.