【摘要】 目的将人外周血淋巴细胞(PBL)以腹腔转输方法构建具备人免疫环境SCID鼠模型,探讨树突状细胞(DC)疫苗对荷瘤SCID鼠血循环膀胱癌细胞微转移转归的影响。方法体外侵袭法从膀胱癌细胞株T24中筛选具有高转移能力亚群(T24-3);从人外周血中分离PBL,经腹腔注射小鼠(4×107/鼠),1d后,于鼠右后肢内侧皮下接种3×106个T24-3细胞,2、4、5、6周分别检测鼠体内人免疫状态(包括血清IgG水平和CD3+、CD4+、CD8+T细胞数量)及外周血细胞角蛋白20(CK20)mRNA表达;用含10%人AB型血清的RPMI-1640培养基,在rhGM-CSF和rhIL-4存在下,37℃、5%CO2环境下体外诱导培养人外周血单个核细胞(PBMC)来源DC。用冻融法获得的T24-3细胞裂解上清液致敏DC;并干预荷人瘤细胞的SCID鼠自发血循环肿瘤细胞微转移,观察大体情况、外周血CK20mRNA表达及瘤体MMP-7mRNA表达。结果SCID鼠于接种T24-3肿瘤细胞后5周进行DC疫苗干预。DC疫苗治疗组小鼠外周血中CK20mRNA表达均阴性,无远处转移发生;PBS处理组小鼠外周血中有2例CK20mRNA表达阳性,3例发生远处转移;DC治疗组小鼠外周血中有1例CK20mRNA表达阳性,1例发生远处转移。DC疫苗治疗组小鼠瘤组织中MMP-7mRNA表达最低,较PBS处理和DC治疗组差异有显著性意义(P<0.01)。结论DC疫苗可以有效控制荷人膀胱癌SCID鼠血循环肿瘤细胞微转移发生,进而延缓或阻止肿瘤转移。更多还原

【Abstract】 Objective To study the effect of dendritic cells loaded with whole tumor antigen on hematogenous micrometastasis of bladder cancer model in hu-PBL-SCID mice.Methods T24-3 cell was selected from human bladder transitional cell carcinoma T24 cell line by Boyden chamber system.The SCID mice were intraperitoneally injected with 4×107 hu-PBL and subcutaneously injected with 3×106 T24-3 and were named hu-PBL-T24-3-SCID model.Human IgG level in the blood plasma of mice was detected by ELISA,and human CD3+,CD4+,CD8+T cells in blood and spleen cells of mice were detected by flow cytometry(FCM)analysis to study human immune reconstruction.Human CK20 mRNA expression in mice peripheral blood was detected by RT-PCR to investigate metastasis of tumor cells.The peripheral blood mononuclear cells(PBMCs)were isolated from human peripheral blood,and were induced into DCs by co-culture with rhGM-CSF and rhIL-4 in vitro.The DC vaccines were produced by co-culturing with whole tumor antigen which was purified through freezing and melting T24-3 cell line.After T24-3 cell was injected into SCID mice for 5 weeks,the mice were treated with DC vaccine.Results All mice were treated from the 5th week on.The expression of CK20 mRNA in peripheral blood of DC vaccine treated mice was the lowest.There were 2 mice showing CK20 mRNA expression and 3 mice with metastatic tumor in PBS group.MMP-7mRNA expression in tumor tissue of DC vaccine treated mice was statistically lower than that in the PBS group(P<0.01).Conclusion DC vaccines have good effects in hu-PBL-SCID mice bladder cancer model by reducing hematogenousmicrometastasis.更多还原