不同磷酸化合物扩增外周血γδ T细胞的效率及条件优化

发布时间：2018-01-04 04:27

本文关键词：不同磷酸化合物扩增外周血γδ T细胞的效率及条件优化　出处：《细胞与分子免疫学杂志》2014年08期 　论文类型：期刊论文

【摘要】：目的研究异戊烯焦磷酸(IPP)与帕米磷酸钠(PAM)体外扩增成人外周血γδT细胞的效率及条件优化。方法梯度离心法分离人外周血单个核细胞(PBMC),分别加入(1.0、5.0、10.0、15.0)μg/mL IPP或(2.0、5.0、8.0、12.0)μg/mL PAM及(100.0、200.0、500.0)IU/mL IL-2,置于100 mL/L小牛血清的RPMI1640培养体系培养,观察并收集细胞,采用流式细胞仪检测刺激后CD3+TCRδ2+的γδT细胞数量以及比例,并计算其扩增效率。结果经过14 d扩增培养后,IPP组的γδT细胞数量在淋巴细胞中比例可上升为81.3%,PAM组的γδT细胞数量在淋巴细胞中比例上升为78.5%,表明IPP和PAM均能有效刺激扩增γδT细胞,2组的扩增效率无统计学差异(P0.05)。结论 PAM具有与IPP相似的体外扩增γδT细胞的能力,PAM扩增法可能成为扩增γδT细胞更为经济实用的选择。
[Abstract]:Objective to study the efficiency and conditions of in vitro amplification of 纬未 T cells from adult peripheral blood by IPP and PAM. Methods Human peripheral blood mononuclear cells (PBMC) were isolated by gradient centrifugation. PBMC). Addition of 1.0 渭 g / mL IPP or 2.0 / 5.0 渭 g / mL / mL PAM and 100.0 渭 g / mL PAM, respectively. The cells were observed and collected in the RPMI1640 culture system of 100 mL/L calf serum. The number and proportion of 纬未 T cells of CD3 TCR 未 2 were measured by flow cytometry. The percentage of 纬未 T cells in lymphocytes in IPP group increased to 81.3%. The percentage of 纬未 T cells in IPP group increased to 78.5%. The results showed that both IPP and PAM could stimulate and amplify 纬未 T cells effectively. There was no significant difference in amplification efficiency between the two groups (P 0.05). Conclusion PAM has the ability to amplify 纬未 T cells in vitro similar to that of IPP. PAM amplification may be a more economical and practical method to amplify 纬未 T cells.
【作者单位】：成都市血液中心;四川大学华西临床医学院;四川大学华西基础医学与法医学院;
【基金】：国家自然科学基金(81170606) 四川大学大学生创新创业计划(201310610068)
【分类号】：R329.2
【正文快照】： γδT细胞是T细胞受体(T cell receptor,TCR)为γδ链的具有独特生物学特征的一类T淋巴细胞亚群[1-2]。γδT细胞不仅有高效的抗病毒和吞噬等典型的天然免疫功能,同时还可以作为特殊的抗原提呈细胞(antigen presenting cell,APC)进一步活化适应性免疫应答[3-4],因而γδT细胞

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