树突状细胞的体外培养及其诱导的抗肿瘤免疫作用

In vitro generation of human blood dendritic cells for active tumor immunotherapy

目的 建立体外培养扩增树突状细胞 (DC)的方法 ,并观察其诱导的抗肿瘤免疫作用 .方法 分离外周血单个核细胞 ,应用 GM- CSF 及 IL- 4诱导 DC产生 ,用 TNF- α和PGE2 促进其成熟 ,并用肺癌细胞系 GL C- 82可溶性抗原致敏 .将成熟 DC与自体 L AK细胞混合培养 (DC- L AK) ,用MTT法检测 DC- L AK细胞及 L AK细胞对多种肿瘤细胞系的杀伤作用 .结果 经 GM- CSF,IL - 4,TNF-α,PGE2 作用后 ,细胞表达 CD1 a阳性率为 71.0 % ,CD83 5 0 .0 % ,CD1 4 阳性率低于 10 .0 % ,高表达 HL A- ,HL A- 和 CD45,为典型的 DC表型特征 .DC- L AK和 L AK细胞对 4种肿瘤细胞 (GL C- 82 ,A5 49,m oser和 MCF- 7)的杀伤率 ,前者为 84.7% ,6 6 .9% ,49.3%和 5 6 .0 % ,后者为 43.5 % ,31.3% ,45 .0 %和 32 .4% .结论 成功的诱导出具有典型形态特征及增强 L AK细胞杀伤活性的 DC.

AIM To establish a method to culture dendritic cells (DC) in vitro and to observe the anti tumor effect induced by DC. METHODS Dendritic cells were derived from PBMC stimulated with GM CSF and IL 4 and DC were pulsed with tumor soluble antigen abstracted from lung can cer cell line GLC 82. TNF α and PGE 2 were used to enhance antigen presenting ability of DC. Phenotype of DC were analyzed through FACS. Mature DC were cocultured with LAK cells and the mixed cells were named DC LAK. Cytotoxicity of DC LAK and LAK were measured by MTT assay. RESULTS The cultured DC expressed HLA Ⅰ, HLA Ⅱ and CD 45 and positive rates of CD 1a, CD 83 and CD 14 were 71.0 %, 50.0% and less than 10.0% respectively. Killing rates of DC LAK and LAK to the four cell lines (GLC 82, A549, moser and MCF 7) were 84.7%, 66.9%, 49.3%, 56.0% and 43.5 %, 31.3%, 45.0%, 32.4% respectively. CONCLUSION DC that have typical phenotype and could be enhance cytotoxity of LAK could be induced from PBMC successfully.