摘要
目的:研究负载LNCaP冻融抗原的树突状细胞(DC)诱导的细胞毒性T淋巴细胞(CTL)对前列腺癌LN-CaP细胞的杀伤效应。方法:采用反复冻融法获得LNCaP细胞抗原,联合应用rhIL-4、rhGM-CSF和TNF-α体外诱导人外周血单个核细胞为成熟DC,并用LNCaP细胞冻融抗原致敏,用该致敏DC体外诱导CTL,观察CTL对LNCaP细胞的特异性杀伤作用,并采用ELISA法检测致敏DCIL-12p70的分泌量。以未致敏DC为对照。结果:冻融抗原致敏DC诱导的CTL对LNCaP细胞具较高杀伤率,该作用强于未致敏DC,且当效靶比为20:1时效用最显著(F组别=68.314,F效靶比=7.251,F交互=56.233,P<0.05);与未致敏DC相比,致敏DCIL-12p70分泌量显著增加(t=4.020,P=0.002)。结论:LNCaP冻融抗原致敏DC激活的CTL在体外具有更强的杀伤LNCaP细胞的作用。
Aim：To investigate the immune responses of cytotoxic T lymphocytes（CTL） against prostate cancer cell LNCaP after activated by dendritic cell（DC） loaded with LNCaP frozen-thawing antigen in vitro.Methods：LNCaP cells were frozen and melt repeatedly 3 times,then tumor lysate was obtained.After separated from heparinized venous blood of healthy donors by Ficoll-Hypaque density-gradient centrifugation,peripheral blood mononuclear cells（PBMCs） were cultured and induced by rhIL-4 and rhGM-CSF and TNF-α to differentiate into DC.The cytotoxicity of CTL activated by DCs loaded with frozen-thawing antigen and cytokine release were measured by MTT assay and ELISA.Results：DCs could be harvested through healthy donors＇ PBMCs cultured in vitro.CTL activated by lysate-loaded DCs had more evident cytotoxicity against LNCaP cells in vitro（Fgroup=68.314,Fratio=7.251,Finteraction=56.233,P0.05）.The level of DCs excreting IL-12,which sensitized by LNCaP lysate,was higher than that of unsensitized DCs（t=4.020,P=0.002）.Conclusion：CTL activated by DCs loaded with LNCaP frozen-thawing antigen exerts a remarkable killing activity on LNCaP cell in vitro.
出处
《郑州大学学报（医学版）》
CAS
北大核心
2012年第6期824-826,共3页
Journal of Zhengzhou University(Medical Sciences)
基金
河南省卫生厅科技攻关基金资助项目200903134
