肺腺癌恶性胸水中肿瘤浸润T淋巴细胞成分及其对自体肿瘤细胞的杀伤活性

Tumor infiltrating T lymphocyte components in malignant pleural effusion of lung adenocarcinoma and their killing activities to autologous tumor cells

目的:分析恶性胸水中的肿瘤浸润T淋巴细胞(tumor infiltrating T lymphocyte,TIL)的细胞成分,探究其对自体肿瘤细胞的杀伤活性。方法:收集17例肺腺癌患者的胸水,采用Ficoll密度梯度离心法分离恶性胸水中的单个核细胞,流式细胞检测仪分析TIL细胞成分,贴壁培养法分离肿瘤细胞和TIL,裸鼠成瘤实验明确贴壁细胞为肿瘤细胞,MTT法检测培养的TIL对自体肿瘤细胞的杀伤活性。结果:恶性胸水TIL中T细胞占60.6%~79.3%,辅助性T细胞占比显著高于细胞毒性T细胞(36.63%±1.90%vs 24.64%±2.32%,P<0.001),同时还含有少量自然杀伤(natural killer,NK)细胞和NK T细胞;采用贴壁培养法成功分离培养肿瘤细胞,体外培养增殖的TIL对自体肿瘤细胞杀伤活性为39.14%±12.04%,且体外高增殖组的TIL对自体肿瘤细胞的杀伤活性高于低增殖组(50.51%±3.80%vs 29.04%±5.77%,P<0.001)。结论:肺腺癌恶性胸水中TIL主要以T细胞为主,辅助性T细胞多于细胞毒性T细胞,体外增殖能力强的TIL对自体肿瘤细胞的杀伤活性高于增殖能力弱者,恶性胸水可能为肿瘤细胞免疫治疗提供细胞来源。

Objective:To analyze the components of tumor infi ltrating T lymphocyte(TIL)cells in malignant pleural eff usion of lung adenocarcinoma,and evaluate their killing activities to autologous tumor cells.Methods:Malignant pleural effusions were collected from 17 patients with lung adenocarcinoma.Mononuclear cells were isolated by Ficoll density gradient centrifugation and flow cytometer was used to analyze TIL cell components.TIL and tumor cells were separated through adherent culture.The tumor cells were identified via intramuscular injection of adherent cells into nude mice and the killing effect of cultured lymphocytes on autologous tumor cells was studied.Results:Of the TIL in malignant pleural effusions,T cells accounted for 60.6%–79.3%,while T helper cells were significantly higher than T killer cells(36.63%±1.90%vs 24.64%±2.32%,P<0.001).There were also natural killer(NK)cells and NK T cells in the effusions.Tumor cells were successfully isolated and cultured.The killing activity of cultured TIL to autologous tumor cells was 39.14%±12.04%,and the killing activity of TIL with high proliferation rate to autologous tumor cells was higher than that of low proliferation group(50.51%±3.80%vs 29.04%±5.77%,P<0.001).Conclusion:T lymphocytes are the major components of TIL in malignant pleural effusions derived from lung adenocarcinoma,and T helper cells are more than T killer cells.The killing activity of TIL with strong proliferation ability to autologous tumor cells is higher than that of TIL with weak proliferation ability.Therefore,cells from malignant pleural effusions could be used for cellular immunotherapy against tumor.