摘要
目的探究阻断程序性死亡受体-1／程序性死亡受体配体1（PD-I／PD-L1）信号通路在骨肉瘤的细胞免疫中的作用及其机制。方法构建裸鼠Saos-2骨肉瘤肺转移模型，Westernbolt法检测移植瘤PD-1和PD-L1表达；使用PD-L1的单克隆抗体给药骨肉瘤模型裸鼠，并以注射溶剂作为对照组，记录裸鼠存活时间、肺转移肿瘤数目和体积；从两组裸鼠肺组织中分离细胞毒性T淋巴细胞（CTL）细胞，与Saos-2细胞共培养，检测给药组和对照组中肿瘤坏死因子（TNF）和炎症因子白细胞介素（IL）-2、IL4的表达量。结果骨肉瘤肺转移模型中PD-1和PD-L1表达高于正常组织，差异具有统计学意义（F=38．93，F=62．38，均P〈0．01）；使用PD-L1的单克隆抗体阻断PD-1和PD-L1相互作用显著提高模型裸鼠存活时间，差异具有统计学意义（F=23．72，P〈0．05）；且给药组肺转移瘤数为（8±4）个，对照组肺转移瘤数为（47±24）个，差异具有统计学意义（F=39．36，P〈0．01），给药组肺转移瘤体积为（32．72±18．75）mm^3，对照组体积为（3．54±2．78）mm^3，差异具有统计学意义（F=48．63，P〈0．01）。对照组CTL的TNF＋、IL-2＋、IL4＋细胞比例分别为（0．63±0．58）％、（0．65±0．47）％、（0．57±0．34）％，而给药组CTL的TNF^＋、IL-2＋、IL4＋细胞比例为（6．84＋1．72）％、（4．63＋0．87）％、（5．23±1．15）％，差异具有统计学意义（F=56．25；F=42．74；F=47．31，均P〈0．01）。结论阻断PD-1／PD-L1信号通路能延缓骨肉瘤恶化，其作用机制与提高CTL细胞对骨肉瘤的细胞免疫潜能有关。
Objective To explore the role of the PD-1/PD-L1 signal pathway blocking in osteosarcoma cellular immunity and their significance. Method the pulmonary metastasis model of Saos-2 osteosareoma in mice were established and western bolt was used to detected the protein expression of PD-1 and PD-L1 ; PD-L1 antibody were administered to osteosarcoma mice, injected solvent were also administered as control, and the mice survival time, number and size of lung metastases tumor was recorded ; cytotoxic T lymphocyte cell of lung tissue of two groups was isolated to co-culture with Saos-2 cells, the expression level of tumor necrosis factor （TNF） and inflammatory cytokines IL-2,IL-4 was detected. Results The expression level of PD-1 and PD-L1 in pulmonary metastasis model of osteosarcoma were significantly higher than normal tissues （ F = 38.93 ; F = 62. 38 ,both P 〈 0. 01 ） ; the use of PD^-＇L1 monoclonal antibody blocking PD-1 and PD-L1 interaction signifi- cantly increased the survival time of osteosarcoma mice（ F = 23.72 ,P 〈 0. 05 ） ; and the pulmonary metastases tumour number of treated groups were （ 8 ＋ 4）, the pulmonary metastases tumour number of control groups were （47 ±24 ） （ F = 39.36, P 〈 0. 01 ）, the pulmonary metastases tumour volume of treated groups were （ 32.72 ± 18.75 ） mm3, volume of control group （ 3.54 ±2.78 ） mm3 （ F = 48.63, P 〈 0. 01 ）. The proportion of TNF ＋ IL-2 ＋ ,IL-4 ＋ CTL in the control group was （ 0.63 ＋ 0.58 ） %, （ 0.65 ＋ 0.47 ） %, （ 0.57 ＋ 0.34 ） % , but in treated group was （6.84 ＋ 1. 72 ） %, （4.63 ＋ 0.87 ） %, （ 5.23 ＋ 1.15 ） % , and the difference was statisticallysignificant （F =56. 25 ;F =42. 74;F =47.31 ,all P 〈0. 01 ）. Conclusion Blockade of PD-1/PD-L1 Signaling Pathway can prolong the deterioration of osteosarcoma, which is associated with the enhancement of CTL immune potential to osteosarcoma.
出处
《国际免疫学杂志》
CAS
2015年第6期537-540,共4页
International Journal of Immunology
