摘要
目的 探讨重组人IL-24联合CIK细胞体外杀伤黑色素瘤A375细胞的效果及其相关作用机制.方法 从黑色素瘤患者外周血分离外周血单个核细胞（PBMC）,通过IFN-γ、IL-2、IL-1α、CD3 McAb诱导培养获得CIK细胞.流式细胞仪检测CIK细胞免疫表型,LDH释放法检测IL-24联合CIK细胞对黑色素瘤A375细胞的杀伤效果.进一步通过检测CIK细胞NKG2D、穿孔素表达和IFN-γ分泌情况,以及A375细胞表面MICA/B的表达水平的,分析IL-24联合CIK细胞对A375细胞杀伤作用机制.结果 流式细胞仪检测结果示CIK细胞中主要效应细胞CD3＋ CD8＋双阳性细胞占（53.0±4.6）％、CD3＋ CD56＋双阳性细胞占（21.5±5.2）％.LDH释放法检测结果示IL-24联合CIK细胞对黑色素瘤A375细胞的杀伤作用最强,20∶1的效靶比时杀伤率达55％.IL-24与CIK细胞联合作用后,CIK细胞表达NKG2D和穿孔素水平提高,IFN-γ的分泌增多;同时IL-24能够提高A375细胞MICA/B的表达.结论 IL-24和CIK细胞联合作用明显增强了CIK细胞对黑色素瘤A375细胞的杀伤能力,其作用机制与IL-24促进CIK细胞分泌IFN-γ和上调NKG2D和穿孔素表达、同时上调A375细胞MICA/B的表达等密切相关。
Objective To investigate the inhibitory effects of combined use of human recombinant IL24 and CIK cell cultures on melanoma A375 cells and its underlying mechanism.Methods Peripheral blood mononuclear cells （PB-MCs） were isolated from patients with melanoma,and then induced into CIK cells by IFN-γ,IL-2,IL-1α,and CD3McAb.The immunophenotypes of CIK cells were examined by flowcytometry.The inhibitory effect of combined use of IL-24 and CIK cell cultures on A375 cells was evaluated by LDH release assay.Furthermore,the amounts of NKG2D,perforin,IFN-γ in/around CIK cells,as well as the expression of MICA/B in A375 cells were detected.Results Flowcytometric results showed that （53.0 ± 4.6） % of CIK cells were CD3 ＋ CD8 ＋ double positive and （21.5 ±5.2） % were CD3 ＋ CD56 ＋ double positive on day 14.According to LDH release assay,the combined use of IL-24 and CIK cell cultures resulted in the strongest killing for A375 cells with a killing rate of 55% at an effector/target ratio of20 ∶ 1.After the combined treatment,CIK cells could produce higher levels of NKG2D,perforin and IFN-γ,while IL-24 promoted the expression of MICA/B in A375 cells.Conclusion Enhanced killing of malignant melanoma A375 cells is observed after combined use of IL24 and CIK cell cultures,which may attribute to the enhanced expression of NKG2D,perforin,IFN-γand MICA/B after IL-24 exposure.
出处
《徐州医学院学报》
CAS
2014年第6期355-359,共5页
Acta Academiae Medicinae Xuzhou
基金
国家自然科学青年基金（81202015）
