摘要
目的:观察单链双特异性抗体(BHL-I)介导的PBL对靶细胞SKOV3的杀伤作用,并研究其细胞毒作用的可能机制。方法:MTT法检测在不同效靶比、不同作用时间、不同靶细胞(SKOV3、BEL-7402)、不同BHL-I浓度等条件下,BHL-I介导PBL对靶细胞的杀伤作用;RT-PCR检测杀伤过程中PBL的穿孔素(Perforin)、颗粒酶(GranzymeB)mRNA的表达及细胞培养上清液中TNF-α、IFN-γ含量变化。结果:BHL-I介导的PBL对SKOV3细胞毒作用明显高于对BEL-7402的,并且在效靶比10∶1、作用36小时、BHL-I浓度为25μg/ml时,PBL对靶细胞SKOV3的细胞毒作用最为显著;在IL-2存在下,BHL-I可引起Perforin、GranzymeB mRNA较高表达及细胞培养上清液中TNF-α、IFN-γ含量升高,当作用时间为72小时时,这些细胞因子的表达有所下降。结论:BHL-I能介导PBL对表达有相应靶抗原的SKOV3细胞具有高效杀伤作用,其细胞毒作用可能与效应细胞表达Perforin、GranzymeB、TNF-α、IFN-γ等有关。
Objective：To investigate the PBL single-strand bispecific antibody BHL-I mediated by killing of SKOV3, and to study on possible mechanisms responsible for the phenomenon. Methods： MTF assay was conducted to measure cytotoxicities of PBL against tumors target cells at various concentration of BHL-I. The content of Perforin, GranzymeB mRNA and of hIFN-γ, hTNF-α secreted by PBL in the process of killing target cells, were evaluated by RT-PCR or ELISA. Results：The cytotoxicity of PBL to SKOV3 was more efficient than to BEL-7402, and the PBL had most effective cytotoxicity to SKOV3 when effector/target ratio was at 10： 1. The optional time and concentration for BHL-I co-incubation were 25 μg/ml and 36 h, respectively. Furthermore, the PBL expressed the higher content of Perforin, GranzymeB mRNA and TNF-α, IFN-γ when IL-2 was present together with BHL-I. Whereas they declined when the time continued for 72 h. Condusion：The BHL-I can induce the PBL efficiently to kill the SKOV3 that has relating Ag. The cytotoxicity activity may be associated with the cytotoxic cytokine（ Perforin, GranzymeB, TNF-α, IFN-γ） which secreted by activated PBL.
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2007年第5期402-406,共5页
Chinese Journal of Immunology
基金
国家重点基础研究发展规划(973)基金资助(No2003CB514113
No2001CB510008)
