【摘要】 目的探讨自体外周血单个核细胞(PBMC)应用于恶性实体瘤过继性免疫治疗的有效性。方法选取恶性实体瘤患者100名,粒细胞动员后,单采PMBC并计数,分析相关因素;流式检测CIK细胞中NKG2D受体率及CIK细胞治疗前后外周血CD3+、CD4+、CD8+、CD4+/CD8+、CD16+CD56+、CD3+CD56+百分率;采用LDH释放法检测PMBC及CIK细胞的杀伤活性;利用Kaplan-Meier生存曲线对比分析CIK治疗组与非治疗组的生存期差异。结果 PMBC数与采血前外周血象的WBC及MNC总数相呈正相关,相关系数r分别为0.62、0.72(P<0.05),而与采集时间及血液流速无关。对照组、实验组各组CIK的杀伤活性与PMBC杀伤活性呈正相关(r分别为0.523、0.627,P<0.05)。培养前、培养至d 7、培养至d 14时CIK细胞杀伤活性都与NKG2D受体表达率成正相关r,分别为0.413、0.5460、.657(P<0.01)。CIK回输治疗前、后CD3+、CD4+、CD8+、CD4+/CD8+、CD16+CD56+、CD3+CD56数量分别为38.66±8.34及62.15±12.36(P<0.05)、30.16±10.26及38.15±9.46(P<0.05)、37.42±11.12及25.74±10.35(P<0.05)1、.15±0.12及1.89±0.12(P>0.05)、17.63±4.25及25.89±8.96(P<0.05)、0.93±0.31及5.67±1.84(P<0.05)。对照组及CIK治疗组的3年生存率平均为86.5%及79%(P<0.05)。结论实体瘤患者的自体PMBC可以用于CIK细胞的制备及治疗。更多还原

【Abstract】 Objective To evaluate the effectiveness of autologous peripheral blood mononuclear cells(PBMC)in adoptive immunotherapy for solid tumor.Methods We collected and counted PMBC of 100 cases of malignant solid tumor patients,mobilized by rhG-CSF,and related factors were analyzed.NKG2D receptors and peripheral CD3+、CD4+、CD8+、CD4+/CD8+、CD16+CD56+、CD3+CD56+ ratio were detected by flow cytometry before and after cytokine-induced killer cells treatment,killing activity of PMBC and CIK were determined by LDH release test,difference of 3-years survival rate between CIK therapy group and non-CIK therapy group were analyzed by Kaplan-Meier survival curve.Results Number of PMBC was correlated positively to white blood cell counts and total circulating blood volume(P<0.05),and unrelated to collection time and blood flow speed.The killing activity of CIK was correlated positively to PMBC in every group(P<0.05).The killing activity of CIK before culture,7 days,and 14 days after culture were all correlated positively to the percentage of NKG2D receptors(P<0.01).The number of CD3+、CD4+,CD8+,CD4+/CD8+,CD16+CD56+,CD3+ CD56+ before and after CIK treatment were 38.66±8.34 vs 62.15±12.36(P<0.05),30.16±10.26 vs 38.15±9.46(P<0.01),37.42±11.12 vs 25.74±10.35(P<0.05),1.15±0.12 vs 1.89±0.12(P>0.05),17.63±4.25 vs 25.89±8.96(P<0.05),0.93±0.31 vs 5.67±1.84(P<0.05),respectively.The mean 3-year survival rate between CIK treatment group and non-treatment group was 86.5% vs 79%(Log-rank test,P<0.05).Conclusion Autoglous PMBC could be used in the preparation of CIK for solid tumor patients.更多还原