肽刺激培养法体外扩增巨细胞病毒特异细胞毒性T淋巴细胞  

Expansion of cytomegalovirus-specific cytotoxic T lymphocytes after peptides stimulation

作　　者：阮光萍[1] 姚翔[1] 安梅[1] 邓淑芬[1] 王桂华[1] 叶蕾[1] 

机构地区：[1]解放军昆明总医院输血科,650032

出　　处：《中华检验医学杂志》2007年第12期1368-1372,共5页Chinese Journal of Laboratory Medicine

摘　　要：目的研制一种扩增特异细胞毒性 T 淋巴细胞频数的方法。方法用不同浓度的巨细胞病毒(CMV)特异的表位肽 pp65刺激人外周血单个核细胞(PBMCs)使 CMV 特异 CTLs 在 PBMCs中的百分比增高。用四聚体-PE 和 CD_8-FITC 双标记 PBMCs 中的 CMV 特异 CTLs,然后用流式细胞仪分析。结果这个方法能有效扩增 CMV 特异 CTLs,使在外周血中占淋巴细胞百分比不到1%的CMV 特异 CTLs 迅速增加到接近20%,占 CD_8^+T 淋巴细胞的40%以上,并能从单份血样中获得大量特异 CTLs 用于流式分析。结论肽刺激培养法简便、易操作,能有效扩增 CMV 特异 CTLs,使不同人的数据更容易对比和连续的评价。该方法也为过继性免疫疗法治疗巨细胞病毒疾病打下了基础。Objective To develop a convenient method efficiently expands the frequency of specific CTLS. Methods We used different concentrations of CMV-specific epitope peptides pp65 to stimulate PBMCs for expansion of CMV-specific CTLs. CMV-specific CTLs were doubly labeled by tetramers-PE and CDs-FITC for FACS analysis. Results The method expands CMV-specific CTLs efficiently. CMV-specific CTLs were expanded from 1% to 20% of PBMCs quickly （namely 40% of CDs T cells）. The method provided a large number of cells with tetramer staining of CDs T cells for FACS analysis from a single blood sampling. Conclusions Peptides stimulation methods are convenient, easy to operate and expanded CMV- specific CTLs efficiently. The increased frequencies of CMV-specific CTLs allowed the data of different individuals to be easily compared and sequentially evaluated. The methods lay the base for adoptive immunotherapy to prevent CMV disease.

关 键 词：T淋巴细胞 细胞毒性 巨细胞病毒 T淋巴细胞 辅助诱导 

分 类 号：R446.6[医药卫生—诊断学]