氩氦冷冻消融对晚期肾癌外周血调节性T 细胞的影响*

目的:研究氩氦冷冻消融治疗对晚期肾癌患者外周血CD4 +CD 25+ 调节性T 细胞（Treg ）的影响及临床意义。方法:回顾性分析本院32例晚期肾癌患者临床资料，所有患者均于氩氦冷冻消融治疗术前、术后1～6 个月分别取外周血，应用流式细胞仪检测外周血T 淋巴细胞亚群（CD3 + T、CD4 + T、CD8 + T、CD4 +T/CD 8 +T、NK细胞、Treg 细胞）变化情况。术后1 个月采用增强MRI或CT评价肿瘤坏死情况，影像学显示肿瘤无强化区即认为肿瘤坏死，根据Cavalieri 方法计算肿瘤坏死率，进而评价肿瘤负荷。结果:冷冻消融治疗术后，Treg 细胞比例由术前（4.18± 1.58）% 逐渐下降至第3 个月达最低值（1.96± 0.54）% ，二者差异有统计学意义（P=0.001）。 CD3 +T、CD4 +T、CD4 +T/CD 8 +T、NK细胞比例分别由术前（19.26± 7.52）% 、（43.54± 12.99）% 、（1.15± 0.57）% 、（17.49±8.36）% 逐渐上升至第3 个月达最大值（30.83± 5.69）% 、（49.58± 10.76）% 、（1.84± 0.12）% 、（27.63± 8.20）% ，差异均有统计学意义（P=0.000，P=0.003，P=0.02，P=0.001）。 CD8 + T由术前（40.86± 8.89）% 逐渐下降至第3 个月达最低值（26.74± 4.29）% ，差异具有统计学意义（P=0.000）。 冷冻治疗后3～6 个月CD3 +T、CD4 +T、CD4 +T/CD 8 +T、NK、CD8 + T、Treg 细胞比例虽略有改变但差异均无统计学意义（P>0.05）。相关性分析结果表明:经氩氦冷冻治疗后Treg 细胞比例变化与肿瘤负荷下降程度呈正相关（r=0.793，P<0.01）。结论:肾癌氩氦冷冻治疗后外周血T 淋巴细胞亚群分布异常得到一定改善，抗肿瘤免疫反应增强，Treg 细胞比例变化与肿瘤负荷有关。 

Effect of Argon-Helium Cryosurgery on Regulatory T Lymphocytes in the Peripheral Blood of Patients with Advanced Renal Carcinoma

Objective: To analyze the effect of Argon-Helium cryosurgery (AHCS) on CD4+ CD25+ regulatory T cells (Treg) and its implication in patients with advanced renal carcinoma.Methods:,Peripheral venous blood samples were ob-tained from 32 patients with advanced renal cell carcinoma before and after AHCS.The proportions of Treg cells and T lym-phocyte subsets (CD3+ T, CD4+ T, CD8+ T, CD4+ T/CD8+ T, and NK cells) in the peripheral blood were measured by flow cytometry.Enhanced CT or enhanced MRI was used to observe the necrosis of tumor at 1 month after AHCS.The areas with no imaging enhancement in tumor were regarded as tumor necrosis.The necrosis rate was measured by Cavalieri method and the tumor burden was evaluated.Results: At 3 months after AHCS, the percentages of Treg cells were gradual-ly decreased from 4.18%±1.58% to 1.96%±0.54%, with a significant difference (P=0.001).At 3 months after AHCS, the pro-portions of CD3+ T, CD4+ T, NK and CD4+ T/CD8+ T were gradually increased from 19.26%±7.52%, 43.54%±12.99%, 1.15%±0.57%, and 17.49%±8.36% to 30.83%±5.69%, 49.58±10.76%, 1.84%±0.12%, and 27.63%±8.20%, with a statistical significance (P=0.000, P=0.003, P=0.02, and P=0.001).The proportion of CD8 + T was decreased from 40.86%±8.89% to the lowest ratio (26.74%±4.29%) at 3 months after AHCS, with a significant difference (P=0.000).At 3～6 months after cryo-therapy, there was only a slight change in the proportions of CD3 + T, CD4 + T, CD4 + T/CD8 + T, NK, CD8 + T, and Treg cells, with no significant difference (P>0.05).Correlation analysis showed that the decrease in tumor burden was positively correlated with the decrease of the proportion of Treg cells (r=0.793, P<0.01).Conclusion: After AHCS, the distribution of T-lymphocyte subsets can be improved and the anti-tumor immune response was strengthened.The percentage of Treg cells is correlated with tumor burden.