摘要
目的:探讨人工抗原提呈细胞(artificial antigen presenting cell,aAPC)K32/4-IBBL/CD86对肝癌CD8+T细胞的活化作用。方法:磁珠法分选HLA-A 2阳性肝癌患者外周血CD8+T细胞,aAPC与CD8+T细胞按不同比例(1∶1、1∶2、1∶3)混合培养,诱导CTL。锥虫蓝拒染法测定CTL的生长曲线,MTS/PMS法检测CTL的增殖,流式细胞术检测CTL分泌IFN-γ的能力,MTT法检测CTL对人肝癌细胞株BEL7402和自体肝癌细胞的杀伤作用。结果:肝癌患者外周血CD8+T细胞经aAPC作用后,增殖能力明显增强,按1∶1、1∶2、1∶3比例混合培养后第8天分别为(21.2±2.5)×105、(17.6±3.2)×105、(15.3±2.8)×105,明显高于对照组的(8.5±0.15)×105(P<0.01),混合培养后分泌IFN-γ的CTL比例分别为(26.2±1.91)%、(21.3±1.38)%、(18.6±1.20)%,明显高于对照组的(0.1±0.02)%(P<0.01)。aAPC活化的CTL对BEL7402细胞和自体肝癌细胞的杀伤率较对照组显著增强,按1∶3混合培养后得到的CTL对BEL7402细胞和自体肝癌细胞的杀伤率分别为(21.8±4.3)%和(25.6±3.6)%,明显高于对照组的(3.8±1.8)%和(3.8±2.0)%(P<0.01);效靶比为50∶1时,1∶1混合培养组CTL对BEL7402细胞的杀伤率(56.8±3.0)%和对自体肝癌细胞的杀伤率(64.8±4.2)%明显高于1∶2混合培养组的(44.3±2.6)%、(56.1±3.4)%和1∶3混合培养组的(38.9±4.7)%、(46.2±4.7)%(P<0.05)。结论:aAPC在体外可高效活化肝癌患者CD8+T细胞,诱导CTL分泌IFN-γ,且CTL对人肝癌细胞株BEL7402和自体肝癌细胞具有明显的杀伤作用。
Objective：To explore the activation of CD8＋T cells of hepatocellular carcinoma（HCC） stimulated by the artificial antigen presenting cells（aAPC）（K32/4-IBBL/CD86）.Methods： Peripheral CD8＋T cells of HLA-A＊0201 positive HCC patients were enriched by MACS separation system.CD8＋T cells were cultured with aAPC at different ratios（3∶1,2∶1 and 1∶1）,and CTLs were induced.Growth curve of CTLs was examined by trypan blue exclusion assay;MTS/PMS assay was used to detect the proliferation of CTLs;the IFN-γ secreting CTLs were assayed by FACS;the cytotoxicity of CTLs on a human liver cancer cell line BEL7402 and self-HCC cells was tested using MTT method.Results： The proliferation of peripheral CD8＋ T cells stimulated with aAPC was significantly increased.At days 8,the CTL cell numbers of aAPC to CD8＋ T cells at 1∶1,1∶2,1∶3 groups was significantly higher than those in the control group（[21.2±2.5]×105,[17.6±3.2]×105,[15.3±2.8]×105 vs [8.5±0.15]×105,P0.01）;and the proportion of IFN-γ secreting CTLs in 1∶1,1∶2,1∶3 groups was higher than those in the control group（[26.2±1.91]%,[21.3±1.38]%,[18.6±1.20]% vs [0.1±0.02]%,P0.01）.The cytotoxicity of aAPC-activated CTLs against BEL7402 cells and self-HCC cells in 1∶3 group was stronger than that in the control group（[21.8±4.3]% vs [3.8±1.8]%,P0.01;[25.6±3.6]% vs [3.8±2.0]%,P0.01）.A higher cytotoxic rate on BEL7402 cells and self-HCC cells in the 1∶1 group at a ratio of E（effect）： T（target） as 50∶1 was achieved compared with that in the 1∶2 and 1∶3 groups（[56.8±3.0]% vs [44.3±2.6]%,[38.9±4.7]%,P0.05;[64.8±4.2]% vs [56.1±3.4]%,[46.2±4.7]%,P0.05）.Conclusion： aAPC can effectively activate CD8＋T cells of HCC patients,and the resultant CTLs not only secrete IFN-γ but also efficiently kill human liver cancer BEL7402 cells and self-HCC cells.
出处
《中国肿瘤生物治疗杂志》
CAS
CSCD
北大核心
2012年第2期148-153,共6页
Chinese Journal of Cancer Biotherapy
基金
江苏省卫生厅预防医学科研基金资助项目(No.Y201032)~~
关键词
人工抗原提呈细胞
肝癌
CD8+T细胞
CTL
IFN-Γ
artificial antigen presenting cell（aAPC）
hepatocellular carcinoma（HCC）
CD8＋T cell
CTL
IFN-γ
