自身γδT细胞联合阿司匹林治疗晚期胃癌的实验研究和临床疗效观察  

Experimental and Clinical Study on Advaced Gastric Cancer Treated with Autologous γδT Cells Combined with Aspirin

作　　者：刘军权[1] 陈复兴[1] 巩新建[2] 李慧忠[1] 蔡凯[1] 张宝福[3] 张颂[1] 张娟[1] 

机构地区：[1]解放军第97医院实验科,徐州221004 [2]解放军第97医院普外科,徐州221004 [3]江苏徐州医学院附属医院泌尿外科,221002

出　　处：《医学研究杂志》2009年第12期32-36,146,共6页Journal of Medical Research

基　　金：南京军区医学科学技术研究"十一五"计划课题资助项目(06MA45)

摘　　要：目的探讨阿司匹林对人γδT细胞功能影响,评估自身γδT细胞联合阿司匹林治疗晚期胃癌的疗效。方法按常规方法培养γδT细胞,收集培养第9天的γδT细胞用不同浓度的阿司匹林诱导24h 后收集上清液用于细胞因子 IFN_(-γ)、TNF—α、IL—12检测,沉淀细胞用于穿孔素、粒酶 B 和 NKG2D 流式细胞测定。17例晚期胃癌患者入院后接受阿司匹林0.1～0.3克/日,口服,3次/日,每4周为1周期,同时采集患者外周血单个核细胞(PBMC),经6～13天培养后分6次输注给患者,回输细胞总数为(2.1～6)×10^(10)个。17例患者中接受1个疗程治疗2例,2个疗程治疗6例,3个疗程治疗6例,4个疗程治疗以上有3例。结果经阿司匹林诱导后γδT细胞穿孔素、粒酶 B、NKG2D 受体表达量明显高于对照组,尤其在阿司匹林浓度为0.8mmol/L时最显著。经0.4～0.8mmol/L 阿司匹林诱导的γδT细胞分泌 IFN_(-γ)量明显高于对照组;分泌的 TNF-α和 IL-12含量与对照组比较无明显差异;当阿司匹林浓度高至3.2mmol/L 时,可以显著抑制 TNF-α的分泌。17例患者用γδT细胞联合阿司匹林治疗后,全组病例均可评价疗效:其中 CR 2例,PR 6例,NC 8例,PD 1例;近期客观有效率47.1%(8/17),中位生存期为12个月。其中有2例患者化疗后接受手术治疗。细胞免疫治疗后大多数患者食欲增加、疼痛减轻、睡眠改善、体重增加。经γδT细胞和阿司匹林联合治疗后患者 CEA 有10例减低,3例增加。结论经阿司匹林诱导后γδT细胞的穿孔素、粒酶 B、NKG2D 受体明显高于对照组,其培养上清液中 IFN_(-γ)含量也明显高于对照组。晚期胃癌经γδT细胞联合阿司匹林治疗后能提高患者生存期,改善患者临床体征,且无毒性不良反应,对晚期胃癌是一种安全有效的治疗方法。Objective To explore the effect of aspirin on human＇ s γδT cell function and to assess the effect of combination of autologous γδT cells with aspirin in advanced gastric cancer. Methods γδT cells cultured routinely were collected on the 9th day and then induced with aspirin in different concentrations. The supernatants were collected after 24h for the cytokines IFN-γ, TNF-α, IL - 12 detection. Sediment cells were detected perforin, granzyme B, NKG2D by flow cytometry （FCM）. A total of 17 cases of advanced gastric cancer after hospitalization received aspirin treatment,0.1-0.3/d, orally, tid. , every four weeks for one cycle. At the same time, peripheral blood mononuclcar cells （PBMC） were collected and cultured for 6 - 13days. The retransfusion cells were infusioned to patients totally 6 times and the total number of the cells were about （2.1-6）×10^10. There were 2 cases received a course of treatment, 6 cases received two courses of treatment, 6 cases received three courses of treatment and 3 cases received more than four courses of treatment in all patients. Results The expression of perforin, granzyme B, NKG2D receptor of γδT cell induced by aspirin were significantly higher than that in the control group, especially in the 0.8mmol/L concentration of aspirin. IFN-γ secretion of γδT cells after induced by aspirin was significantly higher than that in the control group, especially in the 0.4 mmol/L concentration of aspirin. TNF-α and IL- 12 se- cretion of γδT cells had no significant difference as compared with that in the control group. The TNF-α secretion was significantly inhibited when concentrations of aspirin as high as 3.2mmol/L. After being treated with γδT cells combined with aspirin therapy, all patients could be evaluated： CR 3 cases, PR 10 cases, NC 3 cases , PD 1 cases. The recent efficiency objective was 76.5% （13/17） ,and the median survival period was 12 months. There was two cases undergoing surgery after chemotherapy. The majority of patients received cyt

关 键 词：ΓΔT细胞 阿司匹林免疫治疗 穿孔素/粒酶B/NKG2D γ-干扰素/α-肿瘤坏死因子/白介素12 胃癌 

分 类 号：R392.12[医药卫生—免疫学] R971.1[医药卫生—基础医学]