多基因调控的神经胶质细胞瘤瘤苗的制备

Preparation of Multiple Gene Regulated Neuroglioma Vaccine

目的探讨胶质瘤树突状细胞(DC)的融合性混合体瘤苗的制备及其效果。方法经过LipofeetamineTM2000把rIL-12质粒转染给U251细胞、DC和CIK细胞共培养,完成上述两步后,再把U251复合体与DC复合体融合,经过上述过程制备成的胶质瘤DC的融合性混合体瘤苗,该瘤苗经过流式细胞仪OX62+阳性细胞的筛选和纯化,得到提纯的融合瘤苗后,经过体外的混合淋巴细胞反应(MLR)和细胞毒性T淋巴细胞(CTL)测试。结果融合细胞混合体培养72小时见贴壁生长的细胞呈宽大梭形,胞浆丰满,具有较为均一的U251细胞形态。DC复合体与U251复合体细胞按2∶1进行融合,流式细胞学检查见OX62+细胞约占55%,经流式细胞仪分选、OX62+细胞混合体培养72小时,瘤苗的融合效率约为14%。结论自制的DC融合性混合体瘤苗能被分离、存活,呈现出神经胶质瘤细胞形态,胶质瘤融合性瘤苗经Wistar大鼠脾脏来源的MLR反应更强,融合瘤苗活化的CTL对U251的杀伤率明显高于单纯的DC/U251对活化的CTL对U251的杀伤率,具有更强的抗原特异性。

Objective To investigate the preparation and therapeutic effect of glioma-dendritic cells（DC） fusion mixture vaccine. Methods U251 cells was transfected with rlL-12 plasmid by LipofectamineTM 2000 cells was co-cultured with CIK cells. Then the U251 complex was fused with DC complex to prepare glioma-DC fusion mixture vaccine. By OX62＋ screening and purification through cytometry, the purified vaccine was detected by MLR and CTL in vitro. Results The fusion cell complex was wide and full of cytoplasma, with even morphology of U251 cells after 72 h adherent culture. DC complex was fused with U251 complex （2 ： 1）, OX62＋ cell was about 55% assayed by cytometry. Through screening, OX62＋ cell mixture was cultured for 72 h; the fusion rate was about 14%. Conclusion Glioma-DC fusion mixture vaccine prepared in our lab could be separated and survived with displaying neuroglioma cell morphology. The MLR response from spleen of Wistar rats induced by the vaccine was more strong. The kill rate on U251 cells of CTL activated by fusion complex vaccine was higher than that of simple DC/U251 vaccine, suggesting a strong antigen specificity.