一种简单快速扩增和获取外周血γδT细胞的方法

本研究目的是建立一种简单快速的扩增人外周血γδT细胞的方法，以便用于γδT细胞的生物学特性和临床过继性免疫治疗的研究。取人外周血5-10ml，分离获取单个核细胞，用结核杆菌低分子多肽抗原(MTb-Ag)刺激细胞增殖，通过荧光单克隆抗体TCR γδ-PE染色及流式细胞仪检测γδT细胞所占比例，用MTT试验检测细胞毒活性。结果表明：MTb-Ag能特异地刺激外周血单个核细胞中的γδT细胞增殖，γδT细胞可达69.2％，对K562细胞具有较高的杀伤活性。结论：本方法是一种简单、快速、特异的体外扩增和获取人外周血γδT细胞的方法。

A Simple and Rapid Method for Propagation and Purification of the Peripheral Blood γδT cells

The purpose of this study was to set up an approach for expansion of the peripheral blood gammadeltaT cells from normal subjects in order to explore the characteristics of gammadeltaT cells. Peripheral blood mononuclear cells (PBMNC) were separated from 5 - 10 ml peripheral blood and stimulated by the low molecular peptide derived from Mycobacterium tuberculosis (MTb-Ag), and expanded in rIL-2-containing medium. The relative amount of gammadeltaT cells were measured by anti TCR gammadelta-PE staining and flow cytometry. The Cytotoxicity were detected by gammadeltaT assay. The results showed that after stimulation and expansion for 10 days, gammadeltaT cells increased to 69.2% of the total PBMNC and demonstrated significant cytotoxicity against K562 cells. In conclusion, this is a simple, rapid and specific method for expansion of peripheral blood gammadeltaT cells in vitro.