摘要
目的:从人脐血单个核细胞(CBMC)中高效扩增CD56+细胞毒性淋巴细胞。方法:以干细胞生长培养基(SCGM)为基础培养基,设计不同浓度抗CD3单抗、IL鄄2的培养条件,从CBMC中诱导扩增CD56+细胞毒性淋巴细胞,用四甲基氮唑蓝(MTT)法检测其对肿瘤细胞株K562和Raji的杀伤活性,并与抗CD3单抗激活的杀伤细胞(CD3AK)、细胞因子诱导的杀伤细胞(CIK)进行比较。结果:在以SCGM为基础培养基时,CBMC经500ng/ml抗CD3单抗、500U/mlIL鄄2作用获得大量增殖,在12天时扩增(14.7±4.0)倍,(56.4±2.8)%为CD56+细胞,其中CD3-CD56+NK细胞和CD3+CD56+T细胞分别占细胞总数的(24.9±5.3)%和(30.0±4.7)%;在效/靶比10∶1时,对K562和Raji的杀伤率分别为80.5%和55.3%,均显著高于CD3AK和CIK细胞。结论:在抗CD3单抗和IL鄄2作用下可使用SCGM,获得以CD56+淋巴细胞为主的强细胞毒性免疫效应细胞,为进行肿瘤过继免疫治疗提供了一种简单的扩增脐血CD56+细胞毒性淋巴细胞的方法。
Objective: To expand CD56+ cytotoxic lymphocytes from human umbilical cord blood mononuclear cells(CBMC). Methods: CBMC were cultured in stem cell growth medium(SCGM) supplemented with monoclonal anti-CD3 antibodies and IL-2 at varying concentrations to expand CD56+ cytotoxic lymphocytes. The cytotoxicity of the expanded cells against K562 and Raji cell lines was examined by MTT assay and compared with that of CD3AK and CIK cells. Results: After 12 days, CBMC cultured in SCGM supplemented with 500 ng/ml monoclonal anti-CD3 antibodies and 500 U/ml IL-2, expanded(14.7 ± 4.0) fold and contained(56.4 ± 2.8)%CD56+ cells, including(24.9 ± 5.3)% CD3-CD56+NK cells and(30.0 ± 4.7)% CD3+CD56+T cells, respectively. The expanded cell population had significantly higher cytotoxicity than CD3AK and CIK cells, and could lysed 83.7% of K562 and 55.3% of Raji targets at a ratio of 10∶1 between effector and garget. Conclusion: Cytotoxic cells including major CD56+ lymphocytes could be expanded from CBMC using SCGM via the synergistic induction with monoclonal anti-CD3 antibodies and IL-2. This work provides a simple and efficient way of expanding and enriching CD56+ cytotoxic lymphocytes from CBMC for the adoptive immunotheraphy of tumor.
出处
《南京医科大学学报（自然科学版）》
CAS
CSCD
北大核心
2004年第6期612-614,共3页
Journal of Nanjing Medical University(Natural Sciences)
基金
无锡市卫生局"十五"重大科研项目[锡卫科教(2002)3号]
关键词
淋巴细胞
脐血
体外扩增
细胞毒性
lymphocyte
umbilical cord blood
ex vivo expansion
cytotoxicity
