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目的:考察两种载肽聚乳酸((polylaticacid,PLA)免疫微球M1(hAFP158~166)和M2(hAFP218~226)在体外诱导特异性CTL的能力及其对肝癌细胞的杀伤作用。方法:制备分别荷载表位肽hAFP158~166、、hAFP218~226的PLA免疫微球M1和M2,体外刺激HLA-A2+健康志愿者的外周血单个核细胞(PBMC)作为效应细胞,实验分为3组:对照组、hAFP表达组和hAFP阴性组。采用标准51Cr释放法检测CTL杀伤活性。结果:两种免疫微球在体外均能刺激人PBMC增殖,形成大量可见克隆;两者诱导的效应细胞对hAFP+的荷肽T2细胞、HepG-2和Alexander的杀伤率均达75%以上,均显著高于不表达hAFP的膀胱癌细胞BTT和未荷肽的T2细胞,差异非常显著(P<0.01),而两者杀伤活性之间没有明显差异(P>0.05)。结论:两种载肽聚乳酸免疫微球均能在体外诱导产生特异性CTL,并对表达靶抗原的肝癌细胞有较强杀伤作用。
OBJECTIVE: To investigate the ability of two carrier peptides, polylaticacid (PLA) immune microspheres M1 (hAFP158-166) and M2 (hAFP218 ~ 226), to induce specific CTL in vitro and their cytotoxicity on hepatoma cells.Methods: The PBMCs of HLA-A2 + healthy volunteers were stimulated with PLA immunostaining microspheres M1 and M2 containing epitope-bearing peptides hAFP158-166 and hAFP218-226, respectively. The experiment was divided into three groups: Control group, hAFP expression group and hAFP negative group.The CTL killing activity was detected by standard 51Cr release method.Results: Both of the two kinds of immune microspheres could stimulate the proliferation of human PBMC in vitro to form a large number of visible clones.The induced cells induced by hAFP + The killing rates of HepG2, HepG-2 and Alexander reached more than 75%, which were significantly higher than those of BTT and non-peptide-free T2 cells that did not express hAFP (P <0.01) (P> 0.05) .Conclusion: Both of the two peptide-loaded polylactic acid immunospheres can induce specific CTLs in vitro and have a strong killing effect on hepatoma cells expressing the target antigen.