人舌癌Tca8113核外体对树突状细胞诱导CTL杀伤活性的影响

Killing effect of DC-CTL induced by exosomes derived from human tongue cancer Tca8113

目的:探讨人舌癌Tca8113细胞核外体(exosomes)体外诱导外周血来源的树突状细胞(dendritic cell,DC)对CTL杀伤活性的影响。方法:抽取健康成人外周血,分离单核细胞,经贴壁获得树突状细胞前体细胞,在体外先后加入GM-CSF、rhIL-4、LPS和/或exosomes诱导培养9d,获得成熟DC;而未贴壁细胞经rhlL-2诱导培养,获得T淋巴细胞。设置舌癌Tca8113组和对照EC9706组,每组再根据效应细胞不同分为A组:Tca8113细胞核外体致敏的DC与T淋巴细胞共培养组;B组:未经致敏DC与T细胞共培养组;C组:单纯T细胞组。结果:A、B、C组效应细胞对Tca8113细胞株的杀伤活性分别为(78.56±2.26)%、(50.34±6.21)%、(26.47±3.52)%。A与B组、A与C组比较差异有统计学意义(P<0.05),A组效应细胞对EC9706杀伤活性为(51.18±5.31)%,与对Tca8113的杀伤活性相比差异有统计学意义(P<0.05)。结论:人舌癌Tca8113核外体能增强DC-CTL效应细胞对舌鳞癌Tca8113细胞株特异性的杀伤活性。

Objective： To explore the killing effect of CTL. induced by DC from peripheral blood sensitized with exosomes derived from human tongue cancer Tea8113 in vitro. Methods：Tca8113 cells were. cultured successively for 2 days and the supernatant was collected. Exosomes were separated from the supernatant through 0.45um fitter membrane and 100 kDa MWCO Millipore Amicon centrifuging at 5000g. The mononuclear cells collected from normal peripheral blood were cultured with rhIL-4, GM-CSF,LPS and with or without exosomes-to induce DC for 9 days.In this experiment tongue cancer Tea8113 group and control EC9706 group were set up, each group subdivided to gronp A（DC sensitized by exosomes from Tea8113 with T lymphocyte co-cultured）,group B（DC not sensitized by exosomes with T lymphocyte co-cultured）, group C（pure T lymphocyte） group on the hasis of differeutial effector cells. Results：The mononuclear cells were cultured with various cytnkine, typical DCs was observed under phase-contrast microscope and scanning electron microscope.To Tca8113 cell line. the killing activity of A, B, C effector cells were（78.56 ±2.26）% ,（50.34±6.21）% , （26.47 3.52）% respectively.Group A showed significant difference from group B and group C （P〈 0.05）. To EC9706 cell line, the killing activity of group A was （51.18 5.31） %, compared with that of Tea8113, the difference had statistic meaning（P〈0.05）. Conclusion： Tumor freeze thawing antigen derived from human tongue cancer Tea8113 can enhance specific cytotoxieity of DC-CTL