人工抗原呈递细胞复合物体外诱导特异性T淋巴细胞活化的研究  

Activation of Specific T Lymphocyte Induced by Artificial Antigen Presenting Cell Complex In Vitro

作　　者：毛汉文[1] 刘文励[1] 周剑锋[1] 耿哲[1] 黄伟[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院血液科,湖北武汉430030

出　　处：《中国实验血液学杂志》2008年第3期655-658,共4页Journal of Experimental Hematology

基　　金：国家自然科学基金资助项目;编号30400186

摘　　要：本研究旨在以人工抗原呈递细胞(artificial antigen-presenting cells,aAPCs)体外模拟树突状细胞生物学功能,探讨诱导特异性T淋巴细胞活化、增殖的作用。以磁性微珠为载体,选取慢性髓系白血病特异性抗原CML28作为目标抗原肽,通过抗原表位预测软件获取CML28表位序列(Vltfaedsv),并以该抗原表位与MHC分子(HLA-A2-Ig)偶联,作为第一信号分子,B7-1分子作为第二信号分子,将两种信号分子同时负载于磁珠表面,构建人工APC复合体;取HLA-A2+的健康骨髓捐赠者骨髓单个核细胞,以磁珠分选出CD8+T淋巴细胞并与人工APC复合体系统共培养。培养过程中加入羧基荧光素二醋酸盐琥珀酰亚胺酯(5,6-carboxylfluorescin diacetate succinimidylester,CFSE),并以此检测T细胞增殖,用流式细胞仪检测特异性T细胞增殖程度。结果发现:实验组中CML28-特异性T细胞增殖程度明显增高,实验组平均值为(17.34±0.35)%,对照组平均值为(2.25±0.43)%,两者比较差异显著(p<0.01)。结论:人工APC复合体系统在体外能模拟人体抗原呈递细胞,刺激CD8+T特异性淋巴细胞活化、增殖。This study was aimed to investigate the effect of artificial antigen-presenting cells （aAPCs） on inducing activation and proliferation of specific T-lymphocytes through stimulating biological function of dendritic cells with aAPCs in vitro. The specific antigen of chronic myeloid leukemia CML-28 was screened as objective antigen peptide by using magnetic microbeads as vector; the CML-28 epitope sequence （Vlffaldsv） was obtained by antigen epitope prediction software; this epitope was coupled with MHC molecule and used as first signal molecule, the B7-1 molecule was used as second signal molecule; these 2 molecules simultaneously were loaded onto surface of magnetic microbeads so as to contract aAPC complex. The bone marrow monomuclear cells were derived from HLA-A2 ^＋ healthy bone marrow donors, CD8 ^＋ T lymphocytes were screened and co-cultured with aAPC complex. During culture the 5,6- carboxyfluorescein diacetate succinimidyl ester （CFSE） was added and proliferafon of T-lymphocytes was detected by CPSE and proliferation level of specific T lymphocytes was detected by flow cytometry. The results showed that the proliferation level of CML-28 specfic T lymphocytes obviously increased in experimental group, average level was 17.34 ± 0.65 %, while average level in control was 2.25 ±0.43 %, there was significant difference belween them （p 〈 0.01）. It is concluded that the aAPC complex can mimic human APCs in vitro, and stimulate activation and proliferation of CD8^＋ specific T lymphocytes.

关 键 词：人工抗原呈递细胞 特异性T淋巴细胞 树突状细胞 慢性髓系白血病 

分 类 号：R730.51[医药卫生—肿瘤] R733.72[医药卫生—临床医学]