氩氦刀治疗小鼠肺腺癌后瘤周注射树突状细胞对冷冻免疫效应的影响

Effect of Peri-tumoral Injection of Dendritic Cells Following Argon-Helium Crytherapy on Cryoimmune Response in the model of lung adenocarcinoma bearing mice

目的探讨氩氦刀治疗小鼠肺腺癌后,在瘤体周围注射树突状细胞(dendritic cells,DCs)是否可提高冷冻免疫效应。方法 38只T739荷瘤小鼠(LA795肺腺癌,瘤体位于右后肢皮下,直径7±0.5mm)随机分成2组(19只/组)。实验组:双循环氩氦冷冻消融后于瘤周注射DCs;对照组:双循环氩氦冷冻消融。术后第3、7、14d脱颈处死小鼠,眼眶采血(3只/组/时间点),流式细胞术微球阵列法检测细胞因子IL-12,IFN-γ,IL-4,IL-10浓度;流式细胞术检测CD4+T、CD8+T细胞比例;每组10只小鼠用于观察生存时间。结果实验组在各时间点IL-12、IFN-γ浓度均高于对照组,IL-4、IL-10浓度均低于对照组;实验组CD4+T细胞比例在各时间点均高于冷冻治疗组,CD8+T细胞在术后第7、14d均高于对照组。以上所有差异均具有统计学意义(P<0.05)。在术后第3d,实验组CD8+T细胞比例高于对照组,但其差异无统计学意义(P>0.05)。实验组生存率高于对照组,其差异具有统计学意义(P<0.05)。结论氩氦冷冻消融术后在瘤周注射DCs可显著提高冷冻治疗的抗肿瘤免疫效应,并延长生存时间。

Objective To evaluate the effect of peri-tumoral injection of dendritic cells（DCs）following argon-helium cryoablation of lung adenocarcinoma on the cryoimmune response.Methods Thiety-eight LA795 lung adenocarcinoma bearing mice（tumor of 7±0.5mm in diameter was subcutaneous in the right hind femur）were randomly devided into 2groups：experimental group（double-circled Argon-Helium cryoablation peritumoral injection of DCs）and the control group（double-circled Argon-Helium cryoablation）.Mice were excuted on day 3,7and 14 after treatment（3mice/group/time point）for orbital blood sampling.Cytokines（IL-12,IFN-γ,IL-4,IL-10）and T lymphocyte subsets（CD4^＋TCD8^＋T cells）in the blood were tested by Cytometric Bead Array（CBA）flow cytometry（FCM）respectively.Ten mice in each group were used to evaluate survival time.Results Compared with the control group,the experimental group has higher concentration of IL-12 and IFN-γ;while has lower concentration of IL-4and IL-10 at each time point.Percentage of CD4^＋T cells in the experimental group was higher at each time point;percentage of CD8^＋T cells in the experimental group was higher on day 7and 14.All of the differences have statistical significance（P〈0.05）.On day 7post treatment,the percentage of CD8^＋T cells in the experimental group was higher than that in the control group,while the difference showed no statistical significance（P〉0.05）.Survival time of the experimental group was longer than that of the control group,and the difference was statistically significant（P〈0.05）.Conclusion Peri-tumoral injection of DCs following cryotherapy can effectively enhance the anti-tumor croyimmune effect and thus prolong the life span.