树突状细胞诱导高聚金葡素刺激的淋巴细胞体外高效特异性杀瘤作用

High potency and specific tumoricidal activity of HAS-stimulated lymphocytes induced by tumor antigen pulsed dendritic cells

目的体外诱导高效特异的抗肿瘤免疫反应,以期制备新型特异性肿瘤免疫治疗效应细胞。方法分离健康人外周血单个核细胞PBMC将其中非贴壁细胞淋巴细胞加高聚金葡素HAS培养称HASL;贴壁细胞经IL4和GMCSF联合刺激诱导树突状细胞DC分化。DC用人肺癌细胞系GLC82的可溶性抗原致敏,并用TNFα和PGE2促进DC的抗原递呈能力。培养第10天,将DC与经或未经HAS刺激的淋巴细胞混合培养4h,分别称混合培养细胞为DCHASL和DCL。将DCHASL、HASL及DCL作为效应细胞,肿瘤细胞作为靶细胞,用MTT法检测细胞毒活性。细胞表型用流式细胞仪分析。结果DCHASL对GLC82细胞具有高效而特异性的杀伤作用杀伤率100%对另一肺癌细胞系A549细胞的杀伤率也达94.2%对大肠癌细胞moser和乳腺癌细胞MCF7的杀伤率分别为73.5%和71.2%。而HASL和DCL对A549、GLC82、moser和MCF7细胞的杀伤率则分别为48%、63%、60%、45%和42.1%、46.4%、39%、25.6%。淋巴细胞经HAS刺激后CD3+CD56+细胞的比例大幅度增加。结论HAS在体外刺激淋巴细胞增殖产生的HASL对肿瘤细胞有一定的杀伤作用,无明显特异性。而DCHASL则可高效特异性地杀伤GLC82细胞,对其它肿瘤细胞也有不同程度的杀伤作用。另外。

Aim To induce high potency and specific tumoricidal immune response in vitro by HAS stimulated lymphocytes and dendritic cells. Methods Dendritic cells(DC) were derived from PBMC stimulated with GM CSF and IL 4. DC were pulsed with soluble tumor antigen extracted from lung cancer cell line GLC 82 TNF α and PGE2 were used to enhance antigen presenting ability of DC. Nonadherant cells from the PBMC were cultured in the presence of HAS(named HASL).DC and HASL were cocultured for 4 hours on day 10 and the obtained cells were named DC HASL. DC cocultured with lymphocytes were named DC L. Cytotoxicity of HASL,DC L and DC HASL were determined by MTT assay. Phenotypes of DC and HASL were analyzed by FACS. Results Cytotoxic rates of DC HASL to GLC 82 cells reached 100% and the cytotoxic rates to other cancer cell lines A549, moser and MCF 7 were 94.2%,73.5% and 71.2% respectively; Cytotoxic rates of HASL and DC L to the above four cell lines were 63%,48%,60%,45% and 46.4%,42.1%,39%,25.6% respectively. The percent of CD3+CD56+ cells was increasing greatly after stimulated by HAS. Conclusion HASL can lyse tumor cells to a certain extent ,but there was no obvious specificity. DC HASL can specifically lyse tumor antigen bearing GLC 82 cell line and it's cytotoxicity to A549 cells increased distinctly. As compared with DC HASL, DC L has lower cytotoxicity and specificity, suggesting that specific effect cells for tumor immunotherapy can be prepared using DC and HASL,meanwhile, also providing the probability for tumor adoptive immunotherapy by homologous effect cells.