HSP70-TKD诱导的NK细胞对胰腺癌细胞裸鼠移植瘤的抑制作用  

Effect of TKD-activated NK cells on tumor growth inhibition of human pancreatic carcinoma in nude mice

作　　者：陈蓉明[1] 龚福生[1] 郑秋红[1] 谢云青[1] 应敏刚[1] 

机构地区：[1]福建医科大学教学医院福建省肿瘤医院肿瘤分子研究室,福州350014

出　　处：《中国免疫学杂志》2015年第12期1605-1609,共5页Chinese Journal of Immunology

基　　金：福建省医学创新课题(2011-CX-17)

摘　　要：目的:研究HSP70-TKD诱导的自然杀伤细胞(NK)对胰腺癌原位移植模型的体内抗肿瘤作用。方法:用干细胞培养基富集人外周血单个核细胞(Peripheral blood mononuclear cell,PBMC)至第10天时,加入2μg/ml TKD,4 d后用流式细胞仪检测诱导后NK细胞表面杀伤性受体CD94/NKG2C的表达情况,MTT法测定诱导后NK细胞体外对细胞表面HSP70表达阳性的胰腺癌Colo357细胞的杀伤活性。在BALB/c裸鼠胰腺原位接种胰腺癌Colo357细胞至第15天时,尾静脉注射HSP70-TKD诱导的NK细胞,观察NK细胞对荷瘤鼠的抑瘤作用。免疫组化染色检测NK细胞在胰腺癌组织的浸润情况。结果:NK细胞在干细胞培养基中可大量增殖,14 d时平均细胞纯度为(87.50±1.35)%。HSP70-TKD诱导的NK细胞其细胞表面CD94/NKG2C的表达明显增加,其平均荧光强度为(220.56±6.82),高于未诱导组的(68.72±1.85)。HSP70-TKD诱导的NK细胞对胰腺癌细胞Colo357有更强的杀伤活性,当效靶比为40∶1时杀伤率高达(68.72±2.55)%。体内实验表明,TKD诱导的NK细胞能显著抑制胰腺癌的生长,其抑瘤率可达(61.3±1.5)%,与对照组差异显著(P<0.01)。免疫组化染色表明IL-2和TKD同时诱导的NK细胞更易向胰腺癌组织浸润。结论:HSP70-TKD诱导的NK细胞是一种新型、高效的免疫活性细胞,具有较强的体外抗细胞表面HSP70表达阳性的胰腺癌细胞活性,具有明显的抑制胰腺肿瘤生长的作用。Objective： To investigate the antitumor effect of TKD-activated NK cells on tumor growth inhibition of human pancreatic carcinoma in nude mice. Methods： CD3-CD56＋NK cells were obtained from human peripheral blood mononuclear（ PBMC） in stem cell growth medium SCGM,2 μg / ml TKD was added to the medium on day 10. The activating receptor CD94 / NKG2 C expression levels on NK cells was detected with FAC after 4 days. The cytolytic activity of TKD-activated NK cells against human pancreatic carcinoma subline with HSP70 expression on their cell surface was analyzed by MTT assay. Established a new model of orthotopic-transplantation tumor of human pancreas. NK cells were injected i. v. into the tail vein of tumor-bearing mice on day 15,the antitumor activity of the NK were evaluated. The capacity to infiltrate Colo357 tumors in SCID / beige mice was detected with Immunohistochemistry. Results：Flow cytometry analysis showed that CD3-CD56＋NK cells could expanded in SCGM medium,and the average percentage of NK cell was（ 87. 50 ± 1. 35） %. CD94 expression levels on NK cells increased obviously,the mean fluorescence intensity of CD94 was（ 220. 56± 1. 82）,compared to（ 68. 72 ± 1. 85） of control group cell. The cytolytic activity against HSP70 membrane-positive pancreatic carcinoma sublines Colo357 cells was high and there was significantly statistical difference between TKD-activated NK cells and unactivated NK cells. The cytolytic activity was（ 68. 72 ± 2. 55） % when ratio of effector cells and target cell was 40∶ 1. TKD-activated NK cells had a stronger suppressive effect on tumor growth in BALB / c nude mice bearing Colo357 cells in vivo,Median inhibitory rates was（ 61. 3± 1. 5） %. There was significant statistical difference compare to control group（ P〈 0. 01）. The result of Immunohistochemistry indicated that predominantly NK cells induced with TKD had the capacity to infiltrate Colo357 tumors in SCID / beige mice. Conclusion：TKD-activated NK cells are highly efficient cy

关 键 词：胰腺肿瘤 HSP70-TKD NK细胞 CD94 抗肿瘤 

分 类 号：R392.12[医药卫生—免疫学]