不同刺激物对细胞因子诱导的杀伤细胞分泌细胞因子的影响

Influence of different stimulators on secretion of cytokines by cytokine-induced killer cells

目的:探讨抗CD3单抗联合不同浓度的抗CD28单抗对细胞因子诱导的杀伤(CIK)细胞细胞因子分泌的影响。方法:取6例肿瘤患者的外周血,分离、诱导培养,获取CIK细胞。CIK细胞成熟后用1.0 mg/L抗CD3单抗分别联合0.5、1.0、2.5 mg/L抗CD28单抗进行活化,使用ELISA方法检测细胞因子IFN-γ、IL-10、IL-2、TNF-α的分泌水平,并用胞内细胞染色法检测CD3+CD8+T细胞中IFN-γ、穿孔素的表达水平。以50μg/L佛波酯联合750μg/L离子霉素共刺激的CIK细胞作对照。结果:抗CD3单抗联合不同浓度抗CD28单抗组CIK细胞IFN-γ、IL-10、IL-2、TNF-α分泌水平和胞内穿孔素表达水平差异均无统计学意义(P>0.05)。结论:抗CD3单抗联合不同浓度抗CD28单抗共刺激对CIK细胞细胞因子的分泌无影响,临床上可选择1.0 mg/L抗CD3单抗联合0.5 mg/L抗CD28单抗活化淋巴细胞。

Aim：To investigate the effects of anti-CD3 mAb and different concentrations of anti-CD28 mAb stimuli on the secretion of cytokines by cytokine-induced killer（CIK） cells.Methods：CIK cells were obtained from peripheral blood of 6 tumor patients,and stimulated by 1.0 mg/L anti-CD3 mAb combined with 0.5,1.0 or 2.5 mg/L anti-CD28 mAb,re-spectively.The secretion of IFN-γ, IL-10, IL-2 and TNF-αwas measured by ELISA.Flow cytometry was used to detect the expressions of IFN-γand perforin in CIK cells .CIK cells stimulated by 50 μg/L PMA combined with 750 μg/L iono-mycin were the control.Results：There were no statistical differences in the secretion levels of IL-10, IL-2, IFN-γ, TNF-αor the expressions of IFN-γand perforin in CIK cells among the different stimulatory concentrations of anti-CD3 mAb/an-ti-CD28 mAb groups（P＆gt;0.05）.Conclusion：There is no obvious difference in cytokine secretory capacity after stimula-tion with different concentrations of anti-CD3 mAb/anti-CD28 mAb.Thus 1.0 mg/L anti-CD3 mAb and 0.5 mg/L anti-CD28 mAb may be chosen to stimulate lymphocytes .