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目的 研究胆管癌裂解物对转染全长野生型p5 3的树突状细胞 (DC)免疫功能的影响。方法 用腺病毒作为介质 ,将全长野生型 p5 3转染入DC ,然后用胆管癌裂解物修饰已转染全长野生型 p5 3的DC(Lywtp5 3DC) ,检测这种DC表面分子B7-1、B7-2、MHC -Ⅰ、MHC -Ⅱ表达的高低 ,诱导特异的细胞毒性T淋巴细胞 (CTLs)的能力 ,对小鼠的免疫保护和对动物模型治疗作用。结果 经流式细胞仪检测胆管癌裂解物刺激的wtp5 3DC表面高表达B7-1( 86.70± 0 .0 7)、B7-2 ( 18.77± 0 .0 8)、MHC -Ⅰ ( 87.2 0± 0 .0 5 )、MHC -Ⅱ ( 5 6.70± 0 .0 7) ,而单纯DC这些分子是低表达的 ;Lywtp5 3DC能够特异性地杀伤胆管癌细胞 ,杀伤率为 81%。Lywtp5 3DC治疗组和其它组在肿瘤生长的直径大小之间有显著差异 (P <0 .0 5 ) ,在Lywtp5 3DC治疗组中 ,肿瘤生长明显减慢。结论 全长野生型 p5 3基因转染 +胆管癌裂解物联合修饰的DC能诱导CTLs的特异性 ,显著地提高DC的抗原提呈功能 ,体外能诱导高效而特异的抗胆管癌免疫效应。
Objective To study the effect of cholangiocarcinoma lysate on immune function of dendritic cells (DCs) transfected with full-length wild-type p5 3. Methods The full-length wild-type p5 3 was transfected into DC using adenovirus as medium, and the DCs (Lywtp5 3DC) of full-length wild-type p5 3 were transfected with bile duct lysate to detect the expression of B7- 1, B7-2, MHC-I and MHC-II expression, the ability to induce specific cytotoxic T lymphocytes (CTLs), the immunoprotection in mice and the therapeutic effect on animal models. Results The expression of B7-1 (86.70 ± 0.07), B7-2 (18.77 ± 0.08), MHC-I (87.2 ± 0. 0) in wtp5 3DC stimulated by lysates of cholangiocarcinoma were detected by flow cytometry .0 5) and MHC-Ⅱ (5.70 ± 0.07). However, these molecules were low expressed in DC alone. Lywtp5 3DC could specifically kill cholangiocarcinoma cells with a killing rate of 81%. Lywtp5 3DC treatment group and the other groups in the diameter of tumor growth was significantly different (P <0.05), Lywtp5 3DC treatment group, the tumor growth was significantly slowed down. Conclusion The full-length wild type p5 3 gene transfection combined with cholangiocarcinoma lysate modified DC can induce the specificity of CTLs, significantly improve the antigen presenting function of DC, and induce efficient and specific anti-cholangiocarcinoma immune effect in vitro.