胆管癌肿瘤裂解物修饰和P53基因导入对树突状细胞免疫效应的影响

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野生型P5 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% ) ,单纯DC低表达 ;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能诱导细胞毒性T淋巴细胞的特异性 ,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。

Effects of Dendritic Cells Transfected with Full Length Wild Type P53 and Modified by Bile Duct Cancer Lysate on Immune Response

Objective: To investigate the effects of dendritic cells(DCs) transfected with full length wild type P53 and modified by bile duct cancer lysates on immune response. Methods: The wild type P53 was transducted to DCs with adenovirus, and the DCs were modified by bile duct cancer lysates (Lywt-P53DC). The concentration of the surface molecules (B7-1,B7-2 and MHC-Ⅰ,MHC-Ⅱ) of all DCs was detected with FACS, and the ability of the DCs to induce efficient and specific immunological response in anti- 51 Cr-labeled target cells was studied. BalB/c mice were infected with DCs and QBC939, and CTL response in mice immunized with LywtP53DC and treatment of tumor-bearing mice with LywtP53DC and CTL response were studied in those mice. Results: The surface molecules of LywtP53DC had a high expression of B7-1(86.70%±0.07%), B7-2( 18.77% ±0.08% ), MHC-Ⅰ(87.20%±0.05%) and MHC-Ⅱ(56.70%±0.07%) with FACS. The T lymphocytes had a specific CTL lysing ability induced by Lywt-P53DC, and the CTL lysis rate was as high as 81%. The immune protection of LywtP53DC group was obvious, the tumor diameter of LywtP53DC group was (3.10±0.31) mm, (2.73±0.23) mm, (3.70± 0.07) mm on day 13, 16 and 19, smaller than any group of control (P<0.05), DC, wtP53DC and LyDC group. On the other hand, the growth rate of tumor of LywtP53DC group was slower than that of any other group(P< 0.05 ). Conclusion: Dendritic cells transfected with wild type P53 and modified by bile duct cancer lysates have specific CTL killing capability.